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Sample records for bradyrhizobium japonicum infection1cwoa

  1. Co-inoculation effects of Bradyrhizobium japonicum and ...

    African Journals Online (AJOL)

    Co-inoculation effects of Bradyrhizobium japonicum and Azospirillum sp. on competitive nodulation and rhizosphere eubacterial community structures of soybean under rhizobia-established soil conditions.

  2. Gibberellin oxidase activities in Bradyrhizobium japonicum bacteroids.

    Science.gov (United States)

    Méndez, Constanza; Baginsky, Cecilia; Hedden, Peter; Gong, Fan; Carú, Margarita; Rojas, María Cecilia

    2014-02-01

    Bradyrhizobium japonicum bacteroids isolated from root nodules of soybean (Glycine max.) plants converted the gibberellin (GA) precursor [(14)C1]GA12 into several products identified by combined gas chromatography-mass spectrometry as [(14)C1]GA24, [(14)C1]GA9, [(14)C1]GA15, GA9 17-nor-16-one and unidentified products. The oxidation of GA12, catalyzed by the GA 20-oxidase, was present in symbiotic bacteroids from plants around flowering, but not in bacteroids from plants at either an early vegetative stage or at late growth stages. Expression of cps and ks genes, involved in ent-kaurene biosynthesis, was also demonstrated in bacteroids from soybean plants around flowering. Earlier precursors of the GA pathway, ent-[(14)C1]kaurenoic acid or [(14)C4]GA12-aldehyde, were efficiently utilized by B. japonicum bacteroids to give labelled GA9 plus intermediates partially oxidized at C-20, as well as GA9 17-nor-16-one and an unidentified product. No 3β or 13-hydroxylated [(14)C]GAs were detected in any of the incubations. Moreover the C19-GAs [(14)C1]GA4 or [(14)C1]GA20 were recovered unconverted upon incubation with the bacteroids which supports the absence of GA 3β-hydroxylase activity in B. japonicum. The bacterial 20-oxidase utilized the 13-hydroxylated substrates [(14)C1]GA53, [(14)C1]GA44 or [(14)C1]GA19, although with less efficiency than [(14)C1]GA12 to give [(14)C1]GA20 as final product, while the 3β-hydroxylated substrate [(14)C1]GA14 was converted to [(14)C1]GA4 to a very small extent. Endogenous GA9 and GA24 were identified by GC-MS in methanolic nodule extracts. These results suggest that B. japonicum bacteroids would synthesize GA9 under the symbiotic conditions present in soybean root nodules. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Identification of pH tolerant Bradyrhizobium japonicum strains and ...

    African Journals Online (AJOL)

    Eight strains of Bradyrhizobium japonicum were isolated from the root nodules of soybean cultivar PK 472 collected from Adaptive Trial Centre, Bundi, India. All the isolates were authenticated through plant assay test in germination pouches. Growth of the isolated strains on different pH levels was observed and three strains ...

  4. Identification of pH tolerant Bradyrhizobium japonicum strains and ...

    African Journals Online (AJOL)

    Owner

    Eight strains of Bradyrhizobium japonicum were isolated from the root nodules of soybean cultivar PK. 472 collected from Adaptive Trial Centre, Bundi, India. All the isolates were authenticated through plant assay test in germination pouches. Growth of the isolated strains on different pH levels was observed and three ...

  5. A link between arabinose utilization and oxalotrophy in Bradyrhizobium japonicum.

    Science.gov (United States)

    Koch, Marion; Delmotte, Nathanaël; Ahrens, Christian H; Omasits, Ulrich; Schneider, Kathrin; Danza, Francesco; Padhi, Barnali; Murset, Valérie; Braissant, Olivier; Vorholt, Julia A; Hennecke, Hauke; Pessi, Gabriella

    2014-04-01

    Rhizobia have a versatile catabolism that allows them to compete successfully with other microorganisms for nutrients in the soil and in the rhizosphere of their respective host plants. In this study, Bradyrhizobium japonicum USDA 110 was found to be able to utilize oxalate as the sole carbon source. A proteome analysis of cells grown in minimal medium containing arabinose suggested that oxalate oxidation extends the arabinose degradation branch via glycolaldehyde. A mutant of the key pathway genes oxc (for oxalyl-coenzyme A decarboxylase) and frc (for formyl-coenzyme A transferase) was constructed and shown to be (i) impaired in growth on arabinose and (ii) unable to grow on oxalate. Oxalate was detected in roots and, at elevated levels, in root nodules of four different B. japonicum host plants. Mixed-inoculation experiments with wild-type and oxc-frc mutant cells revealed that oxalotrophy might be a beneficial trait of B. japonicum at some stage during legume root nodule colonization.

  6. Evolutionary Instability of Symbiotic Function in Bradyrhizobium japonicum

    Science.gov (United States)

    Sachs, Joel L.; Russell, James E.; Hollowell, Amanda C.

    2011-01-01

    Bacterial mutualists are often acquired from the environment by eukaryotic hosts. However, both theory and empirical work suggest that this bacterial lifestyle is evolutionarily unstable. Bacterial evolution outside of the host is predicted to favor traits that promote an independent lifestyle in the environment at a cost to symbiotic function. Consistent with these predictions, environmentally-acquired bacterial mutualists often lose symbiotic function over evolutionary time. Here, we investigate the evolutionary erosion of symbiotic traits in Bradyrhizobium japonicum, a nodulating root symbiont of legumes. Building on a previous published phylogeny we infer loss events of nodulation capability in a natural population of Bradyrhizobium, potentially driven by mutation or deletion of symbiosis loci. Subsequently, we experimentally evolved representative strains from the symbiont population under host-free in vitro conditions to examine potential drivers of these loss events. Among Bradyrhizobium genotypes that evolved significant increases in fitness in vitro, two exhibited reduced symbiotic quality, but no experimentally evolved strain lost nodulation capability or evolved any fixed changes at six sequenced loci. Our results are consistent with trade-offs between symbiotic quality and fitness in a host free environment. However, the drivers of loss-of-nodulation events in natural Bradyrhizobium populations remain unknown. PMID:22073160

  7. Succinate Dehydrogenase (Sdh) from Bradyrhizobium japonicum Is Closely Related to Mitochondrial Sdh

    OpenAIRE

    Westenberg, David J.; Guerinot, Mary Lou

    1999-01-01

    The sdhCDAB operon, encoding succinate dehydrogenase, was cloned from the soybean symbiont Bradyrhizobium japonicum. Sdh from B. japonicum is phylogenetically related to Sdh from mitochondria. This is the first example of a mitochondrion-like Sdh functionally expressed in Escherichia coli.

  8. Application of earthworm humus and Bradyrhizobium japonicum in Glycine max (L.) Merrill

    OpenAIRE

    Ricardo Gómez Machado; Marta Travieso Torres; Luis Antonio Tamayo López; Yoannia Gretel Pupo Blanco

    2017-01-01

    The experiment was developed in the Granma University's productive area, with the objective of evaluating the alone and combined earthworm humus application effect with Bradyrhizobium japonicum on G7R-315 variety soybean cultivation. Six treatments were evaluated: T1 Control, T2 B. japonicum, T3 earthworm humus (6 t ha-1), T4 B. japonicum + earthworm humus (6 t ha-1), T5 earthworm humus (8 t ha-1), T6 B. japonicum + earthworm humus (8 t ha-1) on a brown soil. A randomized block design was use...

  9. Bioakumulasi Logam Berat oleh Beberapa Galur Bradyrhizobium japonicum

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    ADE NOOR SYAMSUDIN

    2005-09-01

    Full Text Available Heavy metal utilization in industry and agriculture have caused an environmental problem to existing life. Bioaccumulation is made up by a concentration of certain chemical compounds in living tissues. The objective of this research was to reveal the ability of lipopolysaccharides (LPSs of heavy metal Bradyrhizobium japonicum tolerant strains in accumulating heavy metals. The strains used were BDG 10, KDR 10, and KDR 15. The ability of each strains on heavy metal accumulation of Cu, Pb, Zn, Ni, and Cd were quantitatively determined using atomic absorption spectrophotometry. The result showed that each strains has its own capacity to accumulate heavy metals. Accumulation of Cu (0.100, Pb (0.320, and Cd (0.048 ppm/mg dry weight by KDR 10 seem higher than BDG 10 and KDR 15. The highest accumulation of Zn and Ni was shown by KDR 15 in which the value were 0.360 and 0.165 ppm/mg dry weight, respectively and the least accumulation of all heavy metal studied was shown by BDG 10.

  10. Whole-Genome Sequences of 14 Strains of Bradyrhizobium canariense and 1 Strain of Bradyrhizobium japonicum Isolated from Lupinus spp. in Algeria.

    Science.gov (United States)

    Chekireb, Djamel; Crovadore, Julien; Brachmann, Andreas; Chablais, Romain; Cochard, Bastien; Lefort, François

    2017-07-20

    We report here the whole-genome sequences of 14 strains of Bradyrhizobium canariense, isolated from root nodules of Lupinus microanthus and Lupinus angustifolius, and 1 strain of Bradyrhizobium japonicum isolated from root nodules from Lupinus angustifolius in Algeria. These sequences add to the known diversity of this agronomically important genus. Copyright © 2017 Chekireb et al.

  11. Respon Pertumbunan Tanaman Kedelai terhadap Bradyrhizobium japonicum Toleran Masam dan Pemberian Pupuk di Tanah Masam

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    , Triadiati

    2013-10-01

    Full Text Available The use of acid tolerant rhizobacteria such as Bradyrhizobium japonicum is one effort for increasing soybeanproductivity in acid soil. B. japonicum is a N-fixing bacteria that can promote soybean growth through symbiosis with thehost plants. The objective of this study was to investigate the growth and production of soybean var. Wilis inoculated by B.japonicum and NPK inorganic fertilizer application in acid soil. Two isolates of B. japonicum that were BJ 11(19 and BJ11(wt were used as inoculant for soybean. BJ 11(19 was resulted by transposons mutagenesis, whereas BJ 11(wt is a wild type of bacteria. Both isolates of B. japonicum were acid tolerant. Soybean was inoculated with BJ 11(19 and BJ 11(wtcombined with compost and nitrogen fertilizer (with two rates. The field experiment was conducted at Cikabayan, Darmaga,in a randomized complete block design with 12 treatments and 3 replicates. The results showed that application of the acidtolerant B. japonicum BJ 11(wt, compost, and nitrogen fertilizer (10 g m-2 increased the plant height, dry weight of shootsand roots, nodule number, dry weight of nodules, nitrogenase activity, number of pod and seed, seed weight, and nitrogencontent of seeds in acid soil.Keywords: acid soil, acid tolerant rhizobia, Bradyrhizobium japonicum, compost, nitrogen fertilizer

  12. Application of earthworm humus and Bradyrhizobium japonicum in Glycine max (L. Merrill

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    Ricardo Gómez Machado

    2017-10-01

    Full Text Available The experiment was developed in the Granma University's productive area, with the objective of evaluating the alone and combined earthworm humus application effect with Bradyrhizobium japonicum on G7R-315 variety soybean cultivation. Six treatments were evaluated: T1 Control, T2 B. japonicum, T3 earthworm humus (6 t ha-1, T4 B. japonicum + earthworm humus (6 t ha-1, T5 earthworm humus (8 t ha-1, T6 B. japonicum + earthworm humus (8 t ha-1 on a brown soil. A randomized block design was used with three replicas. The evaluated variables were: number per plant leguminous; weigh from 100 seeds and agricultural yield. The data obtained were processed by double classification variance analysis, applied a Turkey's multivariate statistical analysis. It was found that treatments that included the earthworm humus, the evaluated variables shown superior significantly results and it differed of the control treatment and to the aloneB. japonicum application.

  13. Soybean Lectin Enhances Biofilm Formation by Bradyrhizobium japonicum in the Absence of Plants

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    Julieta Pérez-Giménez

    2009-01-01

    Full Text Available Soybean lectin (SBL purified from soybean seeds by affinity chromatography strongly bound to Bradyrhizobium japonicum USDA 110 cell surface. This lectin enhanced biofilm formation by B. japonicum in a concentration-dependent manner. Presence of galactose during biofilm formation had different effects in the presence or absence of SBL. Biofilms were completely inhibited in the presence of both SBL and galactose, while in the absence of SBL, galactose was less inhibitory. SBL was very stable, since its agglutinating activity of B. japonicum cells as well as of human group A+ erythrocytes was resistant to preincubation for one week at 60°C. Hence, we propose that plant remnants might constitute a source of this lectin, which might remain active in soil and thus favor B. japonicum biofilm formation in the interval between soybean crop seasons.

  14. Atmospheric nitrogen uptake by soyabean cultivars in combination with Bradyrhizobium japonicum strains

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    A. D. P Botha

    1997-07-01

    Full Text Available A pot experiment was conducted, using three soyabean (Giycinc max (L. Merrill cultivars (Forrest, Prima and A5409 inoculated at planting, with no (0 and two Bradyrhizobium japonicum strains (WB1 and WB74. The objectives were to investigate the effect of cultivar-rhizobial strain combinations on biomass and N derived from the atmosphere (Ndfa and to compare the atmospheric N uptake determined by the isolope technique, with the conventional method.

  15. Identification and use of actinomycetes for enhanced nodulation of soybean co-inoculated with Bradyrhizobium japonicum.

    Science.gov (United States)

    Gregor, A K; Klubek, B; Varsa, E C

    2003-08-01

    The utilization of actinomycetes as potential soybean (Glycine max (L.)) co-inoculants was evaluated. Soil samples from Carbondale and Belleville, Ill., were used to inoculate pre-germinated soybean plants to determine antibiotic sensitivity in the native Bradyrhizobium japonicum population. Sensitivity was in the order kanamycin > tetracycline > oxytetracycline > rifampicin > neomycin. Antagonism by five actinomycete cultures toward seven test strains of B. japonicum was also assessed. The ranking average inhibition (across all seven B. japonicum strains) by these actino mycetes was Streptomyces kanamyceticus = Streptomyces coeruleoprunus > Streptomyces rimosus > Streptomyces sp. > Amy colatopsis mediterranei. Ten antibiotic combinations were used to isolate antibiotic-resistant mutants of B. japonicum I-110 and 3I1B-110 via successive cycles of mutation. Eighty-one antibiotic-resistant strains were isolated and tested for symbiotic competency; nine of which were selected for further characterization in a greenhouse pot study. Few differences in nodule number were caused by these treatments. Nodule occupancy varied from 0% to 18.3% when antibiotic-resistant strains of B. japonicum were used as the sole inoculants. However, when three mutant strains of B. japonicum were co-inoculated with S. kanamyceticus, significant increases in nodule occupancy (up to 55%) occurred. Increases in shoot nitrogen composition (27.1%-40.9%) were also caused by co-inoculation with S. kanamyceticus.

  16. [Effect of plant growth regulators on physiological activity of Bradyrhizobium japonicum ].

    Science.gov (United States)

    Leonova, N O; Tytova, L V; Tantsiurenko, O V; Antypchuk, A F

    2005-01-01

    Influence of plant growth regulators Ivin, Emistim C, Eney and Agrostimulin on the biomass production and exopolymers synthesis of soybean nodule bacteria, which have contrasting symbiotic properties, and glutamine synthetase activity of their cell-free extracts were studied. It was shown that the processes of the biomass and exopolymers accumulation had an opposite direction. Of all preparations only Ivin and Agrostimulin intensificol growth activity of the microorganisms under study. The level of glutamine synthetase activity and this enzymatic reaction specificity to the bivalent metal ions were determined by the special features of Bradyrhizobium strains and nature of the plant growth regulators. Only in the presence of Eney the increase of glutamine synthetase activity of both cultures of Bradyrhizobium japonicum was established.

  17. Oxygen-dependent catabolism of indole-3-acetic acid in Bradyrhizobium japonicum

    DEFF Research Database (Denmark)

    Egebo, L A; Nielsen, S V; Jochimsen, B U

    1991-01-01

    Some strains of Bradyrhizobium japonicum have the ability to catabolize indole-3-acetic acid (IAA). Examination of this catabolism in strain 110 by in vivo experiments has revealed an enzymatic activity catalyzing the degradation of IAA and 5-hydroxy-indole-3-acetic acid. The activity requires...... addition of the substrates for induction and is oxygen dependent. The highest activity is obtained when the concentration of inducer is 0.2 mM. Spectrophotometric data are consistent with the suggestion that the indole ring is broken during degradation of IAA. We hypothesize that the enzyme catalyzes...

  18. System analysis of salt and osmotic stress induced proteins in Nostoc muscorum and Bradyrhizobium japonicum

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    Vipin Kaithwas

    2017-06-01

    Full Text Available In this study the proteome response of the two diazotrophic organism’s viz. Nostoc muscorum and Bradyrhizobium japonicum exposed to salt (NaCl and osmotic (sucrose stresses was compared. Out of the total over expressed proteins; we have selected only three over expressed proteins viz. GroEL chaperonin, nitrogenase Mo-Fe protein and argininosuccinate synthase for further analysis, and then we analyzed the amino acid frequencies of all the three over expressed proteins. That led to the conclusion that amino acids e.g. alanine, glycine and valine that were energetically cheaper to produce were showing higher frequencies. This study would help in tracing the phylogenetic relationship between protein families.

  19. Complete Genome Sequence of the Soybean Symbiont Bradyrhizobium japonicum Strain USDA6T

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    Nobukazu Uchiike

    2011-10-01

    Full Text Available The complete nucleotide sequence of the genome of the soybean symbiont Bradyrhizobium japonicum strain USDA6T was determined. The genome of USDA6T is a single circular chromosome of 9,207,384 bp. The genome size is similar to that of the genome of another soybean symbiont, B. japonicum USDA110 (9,105,828 bp. Comparison of the whole-genome sequences of USDA6T and USDA110 showed colinearity of major regions in the two genomes, although a large inversion exists between them. A significantly high level of sequence conservation was detected in three regions on each genome. The gene constitution and nucleotide sequence features in these three regions indicate that they may have been derived from a symbiosis island. An ancestral, large symbiosis island, approximately 860 kb in total size, appears to have been split into these three regions by unknown large-scale genome rearrangements. The two integration events responsible for this appear to have taken place independently, but through comparable mechanisms, in both genomes.

  20. Manganese is required for oxidative metabolism in unstressed Bradyrhizobium japonicum cells

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    Hohle, Thomas H.; O’Brian, Mark R.

    2012-01-01

    Recent studies of Mn2+ transport mutants indicate that manganese is essential for unstressed growth in some bacterial species, but is required primarily for induced stress responses in others. A Bradyrhizobium japonicum mutant defective in the high affinity Mn2+ transporter gene mntH has a severe growth phenotype under manganese limitation, suggesting a requirement for the metal under unstressed growth. Here, we found that activities of superoxide dismutase and the glycolytic enzyme pyruvate kinase were deficient in an mntH strain grown under manganese limitation. We identified pykM as the only pyruvate kinase-encoding gene based on deficiency in activity of a pykM mutant, rescue of the growth phenotype with pyruvate, and pyruvate kinase activity of purified recombinant PykM. PykM is unusual in that it required Mn2+ rather than Mg2+ for high activity, and that neither fructose 1,6-bisphosphate nor AMP was a positive allosteric effector. The mntH-dependent superoxide dismutase is encoded by sodM, the only expressed superoxide dismutase-encoding gene under unstressed growth conditions. An mntH mutant grew more slowly on pyruvate under manganese-limited conditions than did a pykM sodM double mutant, implying additional manganese-dependent processes. The findings implicate roles for manganese in key steps in unstressed oxidative metabolism in B. japonicum. PMID:22463793

  1. Discovery of a haem uptake system in the soil bacterium Bradyrhizobium japonicum.

    Science.gov (United States)

    Nienaber, A; Hennecke, H; Fischer, H M

    2001-08-01

    In Bradyrhizobium japonicum, the nitrogen-fixing symbiont of soybeans, we have identified a haem uptake system, Hmu, that comprises a cluster of nine open reading frames. Predicted products of these genes include: HmuR, a TonB-dependent haem receptor in the outer membrane; HmuT, a periplasmic haem-binding protein; and HmuUV, an ABC transporter in the inner membrane. Furthermore, we identified homologues of ExbBD and TonB, that are required for energy transduction from the inner to the outer membrane. Mutant analysis and complementation tests indicated that HmuR and the ExbBD-TonB system, but not the HmuTUV transporter, are essential for haem uptake or haem acquisition from haemoglobin and leghaemoglobin. The TonB system seems to be specific for haem uptake as it is dispensable for siderophore uptake. Therefore, we propose the existence of a second TonB homologue functioning in the uptake of Fe-chelates. When tested on soybean host plants, hmuT-hmuR and exbD-tonB mutants exhibited wild-type symbiotic properties. Thus, haem uptake is not essential for symbiotic nitrogen fixation but it may enable B. japonicum to have access to alternative iron sources in its non-symbiotic state. Transcript analysis and expression studies with lacZ fusions showed that expression of hmuT and hmuR is induced under low iron supply. The same was observed in fur and irr mutant backgrounds although maximal induction levels were decreased. We conclude either that both regulators, Fur and Irr, independently mediate transcriptional control by iron or that a yet unknown iron regulatory system activates gene expression under iron deprivation. An A/T-rich cis-acting element, located in the promoter region of the divergently transcribed hmuTUV and hmuR genes, is possibly required for this type of iron control.

  2. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Tran H.; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese RW; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolic, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-11-11

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e., roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag 8-plex ITRAQ, enriched using Ni-NTA magnetic beads and subjected to nRPLC-MS/MS analysis using HCD and decision tree guided CID/ETD strategy. A total of 1,625 unique phosphopeptides, spanning 1,659 non-redundant phosphorylation sites, were detected from 1,126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5 fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  3. Produtividade da soja em resposta à aplicação de Molibdênio e inoculação com Bradyrhizobium japonicum Soybean yield in response to Molybdenum and Bradyrhizobium japonicum inoculation

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    Eloir Paulo Gris

    2005-02-01

    Full Text Available A soja em simbiose com Bradyrhizobium japonicum é capaz de ter a sua exigência de N satisfeita com a fixação biológica de N2 (FBN. Entretanto, a FBN é afetada pela deficiência Mo, visto que este nutriente faz parte da enzima nitrogenase responsável pelo processo. Foi realizado um experimento, em condição de campo, em Latossolo Vermelho eutroférrico, no município de Palotina, PR, com o objetivo de avaliar o tratamento de sementes com Mo, inoculação de B. japonicum e adubação foliar de Mo na produtividade da soja. Os tratamentos foram quatro doses de Mo (0, 40, 80 e 160 g ha-1 aplicadas em adubação foliar e tratamento com aplicação nas sementes de 40 g ha-1 de Mo, combinados com e sem inoculação com B. japonicum. Não foram observados efeitos estatísticos significativos do tratamento de sementes com Mo, inoculação de B. japonicum e adubação foliar de Mo na produtividade da soja. Não foram obtidas diferenças significativas entre os tratamentos, obtendo-se tendência apra ganho de produtividade apenas com aplicação foliar de 80 g ha-1 de Mo e tratamento de sementes com 40 g ha-1.Soybean in symbiosis with Bradyrhizobium japonicum is able to satisfy its nitrogen (N2 demand with biological nitrogen fixation (BNF. However, BNF can be affected by molybdenium deficiency because this micronutrient is part of the nitrogenase enzyme responsible for the process. An experiment was conducted under field conditions on a Red Latosol (Oxisol in Palotina, state of Paraná, Brazil. The effects of seed treatments with Mo, B. japonicum inoculation as well as foliar Mo fertilization on soybean yield were evaluated. Four Mo rates (0, 40, 80 and 160 g ha-1 were applied as foliar fertilizer in two Mo application forms (without Mo, seed treatment with Mo 40 g ha-1 combined with and without B. japonicum inoculation. There were no significant effects of the seed treatments with Mo, inoculation with B. japonicum and foliar Mo fertilization on

  4. Proteomic Analysis of Soybean [Glycine max (L.) Merrill] Roots Inoculated with Bradyrhizobium japonicum Strain CPAC 15.

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    Torres, Adalgisa R; Rodrigues, Elisete P; Batista, Jesiane Ss; Gomes, Douglas F; Hungria, Mariangela

    2013-01-01

    This research intended to analyze the expression pattern of proteins in roots of the Brazilian soybean cultivar Conquista when inoculated with Bradyrhizobium japonicum CPAC 15, a strain broadly used in commercial inoculants in Brazil. At ten days after bacterial inoculation, whole-cell proteins were extracted from roots and separated by 2-D gel electrophoresis. Comparative analysis revealed significant changes in the intensity of 37 spots due to the inoculation (17 up-regulated and 20 down-regulated proteins), identified by MALDI-TOF/TOF-TOF. Identified proteins were associated with COG functional categories of information storage and processing, cellular processes and signaling, metabolism, and also in the "poorly characterized" and "not in COG" categories. Among the up-regulated proteins, we identified sucrose synthase (nodulin-100), β-tubulin, rubisco activase, glutathione-S-transferase, a putative heat-shock 70-kDa protein, pyridine nucleotide-disulphideoxidoreductase and a putative transposase. Proteomic analysis allowed for the identification of some putative symbiotic functions and confirmed the main biological processes triggered in the nitrogen-fixing symbiosis with soybean.

  5. Mutualistic co-evolution of type III effector genes in Sinorhizobium fredii and Bradyrhizobium japonicum.

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    Jeffrey A Kimbrel

    2013-02-01

    Full Text Available Two diametric paradigms have been proposed to model the molecular co-evolution of microbial mutualists and their eukaryotic hosts. In one, mutualist and host exhibit an antagonistic arms race and each partner evolves rapidly to maximize their own fitness from the interaction at potential expense of the other. In the opposing model, conflicts between mutualist and host are largely resolved and the interaction is characterized by evolutionary stasis. We tested these opposing frameworks in two lineages of mutualistic rhizobia, Sinorhizobium fredii and Bradyrhizobium japonicum. To examine genes demonstrably important for host-interactions we coupled the mining of genome sequences to a comprehensive functional screen for type III effector genes, which are necessary for many Gram-negative pathogens to infect their hosts. We demonstrate that the rhizobial type III effector genes exhibit a surprisingly high degree of conservation in content and sequence that is in contrast to those of a well characterized plant pathogenic species. This type III effector gene conservation is particularly striking in the context of the relatively high genome-wide diversity of rhizobia. The evolution of rhizobial type III effectors is inconsistent with the molecular arms race paradigm. Instead, our results reveal that these loci are relatively static in rhizobial lineages and suggest that fitness conflicts between rhizobia mutualists and their host plants have been largely resolved.

  6. Functional characterization of the Bradyrhizobium japonicum modA and modB genes involved in molybdenum transport.

    Science.gov (United States)

    Delgado, María J; Tresierra-Ayala, Alvaro; Talbi, Chouhra; Bedmar, Eulogio J

    2006-01-01

    A modABC gene cluster that encodes an ABC-type, high-affinity molybdate transporter from Bradyrhizobium japonicum has been isolated and characterized. B. japonicum modA and modB mutant strains were unable to grow aerobically or anaerobically with nitrate as nitrogen source or as respiratory substrate, respectively, and lacked nitrate reductase activity. The nitrogen-fixing ability of the mod mutants in symbiotic association with soybean plants grown in a Mo-deficient mineral solution was severely impaired. Addition of molybdate to the bacterial growth medium or to the plant mineral solution fully restored the wild-type phenotype. Because the amount of molybdate required for suppression of the mutant phenotype either under free-living or under symbiotic conditions was dependent on sulphate concentration, it is likely that a sulphate transporter is also involved in Mo uptake in B. japonicum. The promoter region of the modABC genes has been characterized by primer extension. Reverse transcription and expression of a transcriptional fusion, P(modA)-lacZ, was detected only in a B. japonicum modA mutant grown in a medium without molybdate supplementation. These findings indicate that transcription of the B. japonicum modABC genes is repressed by molybdate.

  7. Polysaccharide-based bioflocculant template of a diazotrophic Bradyrhizobium japonicum 36 for controlled assembly of AgCl nanoparticles.

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    Rasulov, Bakhtiyor A; Pattaeva, Mohichehra A; Yili, Abulimiti; Aisa, Haji Akber

    2016-08-01

    A simple and green method was developed for the biosynthesis of silver chloride nanoparticles, free from silver nanoparticles, using polysaccharide-based bioflocculant of a diazotrophic rhizobacteria Bradyrhizobium japonicum 36 strain. The synthesized silver chloride nanoparticles were characterized by UV-vis, XRD, FT-IR and TEM. The concentration-dependent and controllable method for silver chloride nanoparticles was developed. The biosynthesized silver chloride nanoparticles exhibited strong antimicrobial activity towards pathogenic microorganisms such as Escherichia coli, Staphylococcus aureus and Candida albicans. The synthesized silver chloride nanoparticles can be exploited as a promising new biocide bionanocomposite against pathogenic microorganisms. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Cultivos de Bradyrhizobium japonicum y Sinorhizobium melitoti en medios con hojas de Amaranthus cruentus, como aporte de factores de crecimiento

    OpenAIRE

    Ronchi, Ana L.; Grassano, A. E.; Ripani, G.; Balatti, Antonio Pedro

    2003-01-01

    p.43-50 En este trabajo se estudió la obtención de suspensiones de Sinorhizobium meliloti y Bradyrhizobium japonicum, considerando el efecto de la harina de hoja de amaranto sobre el crecimiento celular en medios recomendados por distintos autores. Los experimentos se realizaron en erlenmeyers en agitador rotatorio a 250 rpm y 2,5 cm de excentricidad. Los estudios realizados permitieron establecer medios de cultivo para alcanzar concentraciones celulares del orden de 2 x 10¹° células viabl...

  9. Nodulação e rendimento de soja co-infectada com Bacillus Subtilis e Bradyrhizobium Japonicum / Bradyrhizobium Elkanii Soybean nodulation and yield when co-inoculated with Bacillus Subtilis and Bradyrhizobium Japonicum / Bradyrhizobium Elkanii

    Directory of Open Access Journals (Sweden)

    Fábio Fernando de Araújo

    1999-09-01

    Full Text Available O Bacillus subtilis pode favorecer o desempenho simbiótico do rizóbio, pelos efeitos na inibição de fitopatógenos ou pela exsudação de fitormônios. Com o objetivo de verificar a viabilidade da co-infecção de sementes de soja com Bradyrhizobium e Bacillus foram conduzidos três experimentos, no Paraná, em solos com população estabelecida de Bradyrhizobium, em que as estirpes de Bradyrhizobium SEMIA 5019 e SEMIA 5080 e suas variantes tolerantes aos metabólitos de Bacillus foram co-infectadas com duas estirpes de Bacillus (AP-3 e PRBS-1, ou seus metabólitos. Na safra 1993/94, em Londrina, o tratamento de co-inoculação de Bradyrhizobium com os metabólitos formulados de Bacillus incrementou, significativamente, em relação ao não-inoculado, o número de nódulos (59%, estádio V3, a ocupação dos nódulos pelas estirpes de Bradyrhizobium (76%, R2 e o rendimento de grãos (24%; em Ponta Grossa, esses incrementos foram de 60%, 145% e 22%, respectivamente. Nessa safra, em Londrina, a co-inoculação das variantes tolerantes com os metabólitos de Bacillus também aumentou o rendimento (26% e N total (17% dos grãos de soja e incrementos significativos foram constatados, na ocupação dos nódulos, pela co-inoculação das variantes tolerantes com as células de Bacillus (78%. Os resultados obtidos indicam a viabilidade da co-inoculação, em sementes de soja, de metabólitos brutos ou formulados ou, ainda, de células de Bacillus subtilis, para incrementar a contribuição do processo de fixação biológica do nitrogênio.Bacillus subtilis can improve rhizobial symbiotic performance by inhibiting plant pathogens or by the exudation of hormones. To verify the viability of co-inoculation of soybean seeds with Bradyrhizobium and Bacillus, three experiments were performed, in the State of Paraná, Brazil, in soils with established population of Bradyrhizobium. The Bradyrhizobium strains SEMIA 5019 and SEMIA 5080, and their natural

  10. Predominant populations of indigenous soybean-nodulating Bradyrhizobium japonicum strains obtained from organic farming systems in Minnesota.

    Science.gov (United States)

    Wongphatcharachai, M; Staley, C; Wang, P; Moncada, K M; Sheaffer, C C; Sadowsky, M J

    2015-05-01

    Bradyrhizobium from organic fields in Minnesota were isolated and genotyped to assess diversity of soybean-bradyrhizobia in organic farming systems that can be used to improve soybean productivity. Soil samples were collected from 25 organic fields in Minnesota during May to July 2012. Soybean (cv. Lambert) was used as a host to trap indigenous bradyrhizobia in each sample. Genetic diversity of Bradyrhizobium strains (n=733) was determined using the horizontal, fluorophore-enhanced, repetitive extragenic palindromic-PCR (HFERP) DNA fingerprinting technique and the soybean-bradyrhizobia were classified into 79 different genotypes. Of these, 15 dominant genotypes were found and were highly similar (>92% fingerprint similarity) to serotypes USDA 127 (40.4%), USDA 4 (31.8%) and USDA 123 (15.5%), which were the three main populations of soybean-bradyrhizobia in organic fields. Bradyrhizobium japonicum serogroup USDA 4 strains were found to make up a previously unrecognized, predominant rhizobial population in the organic farming soils examined. The relative abundance of strain USDA 4 was negatively correlated with that of USDA 127 and this relationship may be influenced by the levels of NO3 -N and other soil edaphic factors. The local community of bradyrhizobia can be affected by applying inoculant bacteria to organic fields. Based on these results, soybean production in organic farms may be improved by displacing strains similar to USDA 4 with those better at nitrogen fixation and competitive ability than indigenous strains. © 2015 The Society for Applied Microbiology.

  11. Catabolism of indole-3-acetic acid and 4- and 5-chloroindole-3-acetic acid in Bradyrhizobium japonicum

    DEFF Research Database (Denmark)

    Jensen, J B; Egsgaard, H; Van Onckelen, H

    1995-01-01

    Some strains of Bradyrhizobium japonicum have the ability to catabolize indole-3-acetic acid. Indoleacetic acid (IAA), 4-chloro-IAA (4-Cl-IAA), and 5-Cl-IAA were metabolized to different extents by strains 61A24 and 110. Metabolites were isolated and analyzed by high-performance liquid...... chromatography and conventional mass spectrometry (MS) methods, including MS-mass spectroscopy, UV spectroscopy, and high-performance liquid chromatography-MS. The identified products indicate a novel metabolic pathway in which IAA is metabolized via dioxindole-3-acetic acid, dioxindole, isatin, and 2......-aminophenyl glyoxylic acid (isatinic acid) to anthranilic acid, which is further metabolized. Degradation of 4-Cl-IAA apparently stops at the 4-Cl-dioxindole step in contrast to 5-Cl-IAA which is metabolized to 5-Cl-anthranilic acid. Udgivelsesdato: 1995-Oct...

  12. Role of Bradyrhizobium japonicum and Trichoderma spp. in the control of root rot disease of soybean

    Directory of Open Access Journals (Sweden)

    Syed Ehteshamul-Haque

    2014-08-01

    Full Text Available Seed treatment of soybean with Bndyrhizobium japonicum, Trichoderma harzianum, T. viride, T. hamatum, T. koningii and T. pseudokoningii significantly controlled the infection of 30-day-old seedlingsby Maerophomina phaseolina, Rhizoctonia solani and Fusarium spp. In 60-day-old plants Trichoderma spp.. and B. japonicum inhibited the grouth of R. solani and Fusarium spp., whereas the use of B. japonicum (TAL-102 with T. harzianum. T. viride, T. koningii and T. pseudokoningii controlled the infection by M. phaseolina. Greater grain yield was recorded when B. japonium (TAI-102 was used with T. hamatum.

  13. Plant recognition of Bradyrhizobium japonicum nod factors. Final report, September 15, 1992--March 14, 1997

    Energy Technology Data Exchange (ETDEWEB)

    Stacey, G.

    1998-01-01

    This grant had three objectives: (1) isolate and identify the unique nod factor metabolites made by different wild-type B. japonicum strains; (2) investigate the biological activity of these unique nod factors, especially as it relates to host range; and (3) initiate studies to define the mechanism of plant recognition of the nod factors. This report summarizes the results of this research.

  14. Transcriptional analysis of genes involved in nodulation in soybean roots inoculated with Bradyrhizobium japonicum strain CPAC 15.

    Science.gov (United States)

    Carvalho, Gesiele Almeida Barros de; Batista, Jesiane Stefânia Silva; Marcelino-Guimarães, Francismar Corrêa; Nascimento, Leandro Costa do; Hungria, Mariangela

    2013-03-06

    Biological nitrogen fixation in root nodules is a process of great importance to crops of soybean [Glycine max (L.) Merr.], as it may provide the bulk of the plant's needs for nitrogen. Legume nodulation involves several complex steps and, although studied for many decades, much remains to be understood. This research aimed at analyzing the global expression of genes in soybean roots of a Brazilian cultivar (Conquista) inoculated with Bradyrhizobium japonicum CPAC 15, a strain broadly used in commercial inoculants in Brazil. To achieve this, we used the suppressive subtractive hybridization (SSH) technique combined with Illumina sequencing. The subtractive library (non-inoculated x inoculated) of soybean roots resulted in 3,210 differentially expressed transcripts at 10 days after inoculation were studied. The data were grouped according to the ontologies of the molecular functions and biological processes. Several classes of genes were confirmed as related to N2 fixation and others were reported for the first time. During nodule formation, a higher percentage of genes were related to primary metabolism, cell-wall modifications and the antioxidant defense system. Putative symbiotic functions were attributed to some of these genes for the first time.

  15. A Putative Type III Secretion System Effector Encoded by the MA20_12780 Gene in Bradyrhizobium japonicum Is-34 Causes Incompatibility with Rj4 Genotype Soybeans.

    Science.gov (United States)

    Tsurumaru, Hirohito; Hashimoto, Syougo; Okizaki, Kouhei; Kanesaki, Yu; Yoshikawa, Hirofumi; Yamakawa, Takeo

    2015-09-01

    The nodulation of Bradyrhizobium japonicum Is-34 is restricted by Rj4 genotype soybeans (Glycine max). To identify the genes responsible for this incompatibility, Tn5 mutants of B. japonicum Is-34 that were able to overcome this nodulation restriction were obtained. Analysis of the Tn5 mutants revealed that Tn5 was inserted into a region containing the MA20_12780 gene. In addition, direct disruption of this gene using marker exchange overcame the nodulation restriction by Rj4 genotype soybeans. The MA20_12780 gene has a tts box motif in its upstream region, indicating a possibility that this gene encodes a type III secretion system (T3SS) effector protein. Bioinformatic characterization revealed that the MA20_12780 protein contains the small ubiquitin-like modifier (SUMO) protease domain of the C48 peptidase (ubiquitin-like protease 1 [Ulp1]) family. The results of the present study indicate that a putative T3SS effector encoded by the MA20_12780 gene causes the incompatibility with Rj4 genotype soybeans, and they suggest the possibility that the nodulation restriction of B. japonicum Is-34 may be due to Rj4 genotype soybeans recognizing the putative T3SS effector (MA20_12780 protein) as a virulence factor. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  16. Effects of the Bradyrhizobium japonicum waaL (rfaL Gene on Hydrophobicity, Motility, Stress Tolerance, and Symbiotic Relationship with Soybeans

    Directory of Open Access Journals (Sweden)

    Jun-Gu Noh

    2015-07-01

    Full Text Available We cloned and sequenced the waaL (rfaL gene from Bradyrhizobium japonicum, which infects soybean and forms nitrogen-fixing nodules on soybean roots. waaL has been extensively studied in the lipopolysaccharide (LPS biosynthesis of enteric bacteria, but little is known about its function in (bradyrhizobial LPS architecture. To characterize its role as O-antigen ligase in the LPS biosynthesis pathway, we constructed a waaL knock-out mutant and its complemented strain named JS015 and CS015, respectively. LPS analysis showed that an LPS structure of JS015 is deficient in O-antigen as compared to that of the wild type and complemented strain CS015, suggesting that WaaL ligates the O-antigen to lipid A-core oligosaccharide to form a complete LPS. JS015 also revealed increased cell surface hydrophobicity, but it showed decreased motility in soft agar plates. In addition to the alteration in cell surface properties, disruption of the waaL gene caused increased sensitivity of JS015 to hydrogen peroxide, osmotic pressure, and novobiocin. Specifically, plant tests revealed that JS015 failed to nodulate the host plant soybean, indicating that the rhizobial waaL gene is responsible for the establishment of a symbiotic relationship between soybean and B. japonicum.

  17. Synthesis of silver nanoparticles on the basis of low and high molar mass exopolysaccharides of Bradyrhizobium japonicum 36 and its antimicrobial activity against some pathogens.

    Science.gov (United States)

    Rasulov, Bakhtiyor; Rustamova, Nigora; Yili, Abulimiti; Zhao, Hai-Qing; Aisa, Haji A

    2016-07-01

    Silver nanoparticles (SNPs) were synthesized on the basis of exopolysaccharides (low and high molar mass) of diazotrophic Bradyrhizobium japonicum 36 strain. The synthesis of SNPs was carried out by direct reduction of silver nitrate with ethanol-insoluble (high molar mass, HMW) and ethanol-soluble (low molar mass, LMW) fractions of exopolysaccharides (EPS), produced by diazotrophic strain B. japonicum 36. SNPs were characterized using UV-vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). SNPs synthesized on the basis of LMW EPS absorbed radiation in the visible regions of 420 nm, whereas SNPs based on the HMW EPS have a wavelength maximum at 450 nm because of the strong SPR transition. Moreover, the antibacterial and antifungal activities of the SNPs were examined in vitro against Escherichia coli, Staphylococcus aureus, and Candida albicans. SNPs synthesized on the basis of LMW EPS were active than those synthesized on the basis of HMW EPS. Besides, UV-visible spectroscopic evaluation confirmed that SNPs synthesized on the basis of LMW EPS were far more stable than those obtained on the basis of HMW EPS.

  18. Effects of the Bradyrhizobium japonicum waaL (rfaL) Gene on Hydrophobicity, Motility, Stress Tolerance, and Symbiotic Relationship with Soybeans.

    Science.gov (United States)

    Noh, Jun-Gu; Jeon, Han-Eul; So, Jae-Seong; Chang, Woo-Suk

    2015-07-23

    We cloned and sequenced the waaL (rfaL) gene from Bradyrhizobium japonicum, which infects soybean and forms nitrogen-fixing nodules on soybean roots. waaL has been extensively studied in the lipopolysaccharide (LPS) biosynthesis of enteric bacteria, but little is known about its function in (brady)rhizobial LPS architecture. To characterize its role as O-antigen ligase in the LPS biosynthesis pathway, we constructed a waaL knock-out mutant and its complemented strain named JS015 and CS015, respectively. LPS analysis showed that an LPS structure of JS015 is deficient in O-antigen as compared to that of the wild type and complemented strain CS015, suggesting that WaaL ligates the O-antigen to lipid A-core oligosaccharide to form a complete LPS. JS015 also revealed increased cell surface hydrophobicity, but it showed decreased motility in soft agar plates. In addition to the alteration in cell surface properties, disruption of the waaL gene caused increased sensitivity of JS015 to hydrogen peroxide, osmotic pressure, and novobiocin. Specifically, plant tests revealed that JS015 failed to nodulate the host plant soybean, indicating that the rhizobial waaL gene is responsible for the establishment of a symbiotic relationship between soybean and B. japonicum.

  19. Labeling of Carbon Pools in Bradyrhizobium japonicum and Rhizobium leguminosarum bv viciae Bacteroids following Incubation of Intact Nodules with CO(2).

    Science.gov (United States)

    Salminen, S O; Streeter, J G

    1992-10-01

    The aim of the work reported here was to ascertain that the patterns of labeling seen in isolated bacteroids also occurred in bacteroids in intact nodules and to observe early metabolic events following exposure of intact nodules to (14)CO(2). Intact nodules of soybean (Glycine max L. Merr. cv Ripley) inoculated with Bradyrhizobium japonicum USDA 110 and pea (Pisum sativum L. cv Progress 9) inoculated with Rhizobium leguminosarum bv viciae isolate 128C53 were detached and immediately fed (14)CO(2) for 1 to 6 min. Bacteroids were purified from these nodules in 5 to 7 min after the feeding period. In the cytosol from both soybean and pea nodules, malate had the highest radioactivity, followed by citrate and aspartate. In peas, asparagine labeling equaled that of aspartate. In B. japonicum bacteroids, malate was the most rapidly labeled compound, and the rate of glutamate labeling was 67% of the rate of malate labeling. Aspartate and alanine were the next most rapidly labeled compounds. R. leguminosarum bacteroids had very low amounts of (14)C and, after a 1-min feeding, malate contained 90% of the radioactivity in the organic acid fraction. Only a trace of activity was found in aspartate, whereas the rate of glutamate and alanine labeling approached that of malate after 6 min of feeding. Under the conditions studied, malate was the major form of labeled carbon supplied to both types of bacteroids. These results with intact nodules confirm our earlier results with isolated bacteroids, which showed that a significant proportion of provided labeled substrate, such as malate, is diverted to glutamate. This supports the conclusion that microaerobic conditions in nodules influence carbon metabolism in bacteroids.

  20. Expression of Animal Anti-Apoptotic Gene Ced-9 Enhances Tolerance during Glycine max L.–Bradyrhizobium japonicum Interaction under Saline Stress but Reduces Nodule Formation

    Science.gov (United States)

    Robert, Germán; Muñoz, Nacira; Melchiorre, Mariana; Sánchez, Federico; Lascano, Ramiro

    2014-01-01

    The mechanisms by which the expression of animal cell death suppressors in economically important plants conferred enhanced stress tolerance are not fully understood. In the present work, the effect of expression of animal antiapoptotic gene Ced-9 in soybean hairy roots was evaluated under root hairs and hairy roots death-inducing stress conditions given by i) Bradyrhizobium japonicum inoculation in presence of 50 mM NaCl, and ii) severe salt stress (150 mM NaCl), for 30 min and 3 h, respectively. We have determined that root hairs death induced by inoculation in presence of 50 mM NaCl showed characteristics of ordered process, with increased ROS generation, MDA and ATP levels, whereas the cell death induced by 150 mM NaCl treatment showed non-ordered or necrotic-like characteristics. The expression of Ced-9 inhibited or at least delayed root hairs death under these treatments. Hairy roots expressing Ced-9 had better homeostasis maintenance, preventing potassium release; increasing the ATP levels and controlling the oxidative damage avoiding the increase of reactive oxygen species production. Even when our results demonstrate a positive effect of animal cell death suppressors in plant cell ionic and redox homeostasis under cell death-inducing conditions, its expression, contrary to expectations, drastically inhibited nodule formation even under control conditions. PMID:25050789

  1. Synthetic Lethality of the bfr and mbfA Genes Reveals a Functional Relationship between Iron Storage and Iron Export in Managing Stress Responses in Bradyrhizobium japonicum.

    Directory of Open Access Journals (Sweden)

    Siva Sankari

    Full Text Available An mbfA mutant of Bradyrhizobium japonicum defective in iron export is sensitive to short term exposure to high levels iron or H2O2. Here, we found that the mbfA strain grown in elevated iron media (100 μM became resistant to those treatments, suggesting a stress response adaptation. The bfr gene encodes the iron storage protein bacterioferritin, and its expression is derepressed by iron. An mbfA bfr double mutant showed a loss of stress adaptation, and had a severe growth phenotype in high iron media. Moreover, a bfrup allele in which bfr is constitutively derepressed conferred stress tolerance on an mbfA mutant without elevating the iron content in the growth media. The intracellular iron content of the mbfA bfr double mutant was substantially higher than that found in the wild type, even when grown in relatively low iron media (5 μM. Under that condition, iron-responsive gene expression was aberrant in the mbfA bfr strain. Moreover, the double mutant was sensitive to the iron-activated antibiotic streptonigrin. We conclude that MbfA and Bfr work in concert to manage iron and oxidative stresses. In addition, the need for iron detoxification is not limited to extreme environments, but is also required for normal cellular function.

  2. Application of AM Fungi with Bradyrhizobium japonicum in improving growth, nutrient uptake and yield of Vigna radiata L. under saline soil

    Directory of Open Access Journals (Sweden)

    Nisha Kadian

    2014-08-01

    Full Text Available A pot experiment was conducted under polyhouse conditions, to evaluate the effect of two different arbuscular mycorrhizal fungi (G. mosseae and A. laevis in combination with Bradyrhizobium japonicum on growth and nutrition of mungbean plant grown under different salt stress levels (4 dS m−1, 8dS m−1 and 12 dS m−1. It was found that under saline conditions, mycorrhizal fungi protect the host plant against the detrimental effect of salinity. The AM inoculated plants showed positive effects on plant growth, dry biomass production, chlorophyll content, mineral uptake, electrolyte leakage, proline, protein content and yield of mungbean plants in comparison to non-mycorrhizal ones but the extent of response varied with the increasing level of salinity. In general, the reduction in Na uptake along with associated increase in P, N, K, electrolyte leakage and high proline content were also found to be better in inoculated ones. The overall results demonstrate that the co-inoculation of microbes with AM fungi promotes salinity tolerance by enhancing nutrient acquisition especially phosphorus (P, producing plant growth hormones, improving rhizospheric and condition of soil by altering the physiological and biochemical properties of the mungbean plant.

  3. Análisis de la interacción soya-cepa (Bradyrhizobium japonicum x ambiente, en oxisoles de la Orinoquia colombiana

    Directory of Open Access Journals (Sweden)

    Valencia Rubén

    2010-11-01

    Full Text Available

    La Orinoquia colombiana y en particular las sabanas de la altillanura, representa una vasta región con ventajas comparativas y competitivas para la producción de soya. La especie se asocia con bacterias del género Bradyrhizobium para suplir, parcial o totalmente, los requerimientos de nitrógeno. Esta asociación es variable y de alta especificidad huésped-rizobio. La investigación tuvo como propósito determinar la interacción existente entre variedades de soya, cepas del género Bradyrhizobium japonicum y ambiente, en oxisoles de la Orinoquia colombiana, para potenciar el efecto de la fijación biológica del nitrógeno (FBN, en la productividad del cultivo. Se evaluaron las variedades Soyica P-34, Orinoquia 3, C. Libertad 4, C. Taluma 5, C. Superior 6 y C. Sabana 7, con las cepas de B. japonicum ICA J-01, J-96, J-98, mezcla de J-01+J-96 y un control con fertilización con nitrógeno de 150 Kg h-1, en ocho de la Orinoquia. La interacción variedad-cepa-ambiente para la variable rendimiento de grano fue determinada mediante los modelos AMMI (Additive main effects and multiplicative interaction y SREG (Sites regression. El rendimiento de grano varió a través de los ambientes, y estuvo influenciado por el tipo de cepa y variedad de soya. Los biplot generados por los modelos AMMI y SREG permitieron identificar la localidad C.I. Libertad 2008A como el ambiente más discriminante y representativo. El ordenamiento de las variedades x cepa y la identificación de mega-ambientes fueron más consistentes con el modelo SREG (r=0.97, ≤ 0,01. En general, las variedades presentaron una respuesta positiva a las cepas J96, mezcla de J-01+J-96 y J98, y una respuesta negativa con J-01 y aplicación de N. Las variedades más estables productivas fueron C. Superior 6 y Orinoquia 3 con la cepa J96. En la interacción, se observó una consistente asociación (r=0

  4. Expressão dos genes nodC, nodW e nopP em Bradyrhizobium japonicum estirpe CPAC 15 avaliada por RT-qPCR Expression of nodC, nodW and nopP genes in Bradyrhizobium japonicum CPAC 15 strain evaluated by RT-qPCR

    Directory of Open Access Journals (Sweden)

    Simone Bortolan

    2009-11-01

    Full Text Available O objetivo deste trabalho foi avaliar a expressão, por RT-qPCR, dos genes de nodulação nodC e nodW e do gene nopP da estirpe CPAC 15, que provavelmente atuam na infecção das raízes da soja. Foram realizados dois experimentos. No primeiro, a expressão dos genes foi avaliada nas células após a incubação com genisteína por 15 min, 1, 4 e 8 horas. Os resultados revelaram que os três genes apresentaram maior expressão imediatamente após o contato com o indutor (15 min. No segundo experimento, a bactéria foi cultivada na presença de indutores (genisteína ou exsudatos de sementes de soja por 48 horas. A expressão dos três genes foi maior na presença de genisteína, com valores de expressão para nodC, nodW e nopP superiores ao controle. Os resultados obtidos confirmam a funcionalidade dos três genes na estirpe CPAC 15, com ênfase para o nopP, cuja funcionalidade em Bradyrhizobium japonicum foi descrita pela primeira vez.The objective of this work was to evaluate, by RT-qPCR, the expression of the nodC and nodW nodulation genes and of the nopP gene of the CPAC 15 strain, which probably play a role in the infection of soybean roots. Two experiments were done. In the first, the gene expression was evaluated in cells after incubation with genistein for 15 min, 1, 4 and 8 hours. Results showed that the three genes showed higher expression immediately after contact with the inducer (15 min. In the second experiment, the bacterium was grown in the presence of inducers (genistein or soybean seed exudates for 48 hours. The expression of the three genes was greater when induced by genistein, and the expression of nodC, nodW and nopP had higher values than the control. The results confirm the functionality of the three genes in the CPAC 15 strain, with an emphasis on the nopP, whose functionality in Bradyrhizobium japonicum was described for the first time.

  5. Iron regulation of gene expression in the Bradyrhizobium japonicum/soybean symbiosis. Final technical report, June 1, 1991--May 31, 1995

    Energy Technology Data Exchange (ETDEWEB)

    Guerinot, M.L.

    1996-02-08

    B.japonicum produces ALA in a reaction catalyzed by the product of the hemA gene. Expression of the gene is affected by iron availability. To address the question of how the 5 prime untranslated region of the hemA transcript is involved in iron regulation, evenly spaced 10bp deletions within the hemA leader region was constructed and effects on hemA-lacZ expression were determined.

  6. Efeito do pH do solo rizosférico e não rizosférico de plantas de soja inoculadas com Bradyrhizobium japonicum na absorção de boro, cobre, ferro, manganês e zinco Effect of pH of rhizospheric and non-rhizospheric soil on boron, copper, iron, manganese, and zinc uptake by soybean plants inoculated with Bradyrhizobium japonicum

    Directory of Open Access Journals (Sweden)

    Luiz Humberto Souza

    2010-10-01

    Full Text Available Alterações no pH da rizosfera de plantas fixadoras de N2 parecem exercer papel fundamental na absorção de micronutrientes que têm sua disponibilidade dependente de alterações da acidez do solo. Estudaram-se variações na absorção de B, Cu, Fe, Mn e Zn durante o ciclo de crescimento e desenvolvimento da soja, induzidas pela fixação biológica de N2 e pelo pH inicial de amostras de dois solos (um LV argiloso e outro arenoso, em um ensaio conduzido em casa de vegetação. Essas amostras foram incubadas com doses de CaCO3 + MgCO3 (4:1 para elevar o pH (H2O a valores de 5,2, 5,6, 6,2 e 6,6 no solo argiloso e 5,3, 5,6, 5,9 e 6,3 no solo arenoso. Após 60 dias de incubação, essas amostras receberam 450 mg dm-3 de P e 120 mg dm-3 de K no solo. Sementes de soja (Glycine max (L Merrill, variedade Paranaíba, inoculadas com Bradyrhizobium japonicum, estirpes SEMIA 587 e SEMIA 5019, foram colocadas para germinar. Foram cultivadas quatro plantas por vaso (2,2 dm³ e colhidas aos 16, 20, 24, 28, 32, 36, 40, 46 e 54 dias após a emergência. Determinaram-se o pH da rizosfera (pHr, o pH do solo entre raízes - não rizosférico (pHnr, os teores de B, Cu, Fe, Mn e de Zn na parte aérea e raiz, o N apenas na parte aérea, o número de nódulos e o peso da matéria seca de parte aérea, raiz e nódulos. Observou-se que as mudanças ocorridas no pHr e pHnr foram dependentes do pH inicial dos solos (pHs e da fixação biológica de N2. O acúmulo de B e de Fe na parte aérea não foi alterado pelos valores de pHr, modificados em função do pHs, exceto para o Fe no solo argiloso. Todavia, aumentos significativos no acúmulo destes nutrientes na parte aérea ocorreram com o aparecimento dos nódulos, a partir de 24 dias após a emergência. Para Cu, Mn e Zn, as diferenças apareceram sobretudo quanto ao pHs. O conteúdo de micronutrientes na planta revelou-se sensível a mudanças no pH rizosférico, principalmente após a nodulação.Changes in the p

  7. Genetic divergence of bradyrhizobium strains nodulating soybeans as revealed by multilocus sequence analysis of genes inside and outside the symbiosis island.

    Science.gov (United States)

    Zhang, Xing Xing; Guo, Hui Juan; Wang, Rui; Sui, Xin Hua; Zhang, Yan Ming; Wang, En Tao; Tian, Chang Fu; Chen, Wen Xin

    2014-05-01

    The genus Bradyrhizobium has been considered to be a taxonomically difficult group. In this study, phylogenetics and evolutionary genetics analyses were used to investigate divergence levels among Bradyrhizobium strains nodulating soybeans in China. Eleven genospecies were identified by sequence analysis of three phylogenetic and taxonomic markers (SMc00019, thrA, and truA). This was also supported by analyses of eight genes outside the symbiosis island ("off-island" genes; SMc00019, thrA, truA, fabB, glyA, phyR, exoN, and hsfA). However, seven genes inside the symbiosis island ("island" genes; nifA, nifH, nodC, nodV, fixA, trpD, and rhcC2) showed contrasting lower levels of nucleotide diversity and recombination rates than did off-island genes. Island genes had significantly incongruent gene phylogenies compared to the species tree. Four phylogenetic clusters were observed in island genes, and the epidemic cluster IV (harbored by Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens, Bradyrhizobium huanghuaihaiense, Bradyrhizobium liaoningense, Bradyrhizobium daqingense, Bradyrhizobium sp. I, Bradyrhizobium sp. III, and Bradyrhizobium sp. IV) was not found in Bradyrhizobium yuanmingense, Bradyrhizobium sp. II, or Bradyrhizobium elkanii. The gene flow level of island genes among genospecies is discussed in the context of the divergence level of off-island genes.

  8. Nodulation and N2 fixation effectiveness of Bradyrhizobium strains in symbiosis with Adzuki Bean, Vigna angularis

    Directory of Open Access Journals (Sweden)

    Dušica Delić

    2010-04-01

    Full Text Available In pot experiment, one isolate Knj from a Serbian soil, four strains of Bradyrhizobium japonicum and three strains of Bradyrhizobium spp. were examined for the effect on adzuki bean nodulation and effectiveness in symbiotic N2 fixation. All the tested strains produced root nodules in adzuki bean. Strains of B. japonicum showed high potential of N2 fixation, particularly 525 and 542. B. japonicum strains resulted 65-71% shoot dry weight and 99-138% total N content of uninoculated control with full N content (100%. No significant difference was found between the plants inoculated with Bradyrhizobium spp. strains and uninoculated control plants without N (40-42 and 42% shoot dry weight, respectively, which indicated symbiotic N2 fixation inactivity of the Bradyrhizobium spp. strains. Knj strain had the middle position (56% shoot dry weight. These data showed that B. japonicum 525 and 542 strains could be used in further investigations in order to apply them as inoculants in microbiological N fertilizers.

  9. Genome sequence of Bradyrhizobium embrapense strain CNPSo 2833T, isolated from a root nodule of Desmodium heterocarpon

    Directory of Open Access Journals (Sweden)

    Jakeline Renata Marçon Delamuta

    Full Text Available Abstract Bradyrhizobium embrapense CNPSo 2833T is a nitrogen-fixing symbiont of the legume pasture Desmodium. Its draft genome contains 8,267,832 bp and 7876 CDSs. The symbiotic island includes nodulation and nitrogen fixation genes resembling the operon organization of B. japonicum. Several CDSs related to secretion proteins and stress tolerance were also identified.

  10. [Queries related to the technology of soybean seed inoculation with Bradyrhizobium spp].

    Science.gov (United States)

    Lodeiro, Aníbal R

    2015-01-01

    With the aim of exploiting symbiotic nitrogen fixation, soybean crops are inoculated with selected strains of Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens or Bradyrhizobium elkanii (collectively referred to as Bradyrhizobium spp.). The most common method of inoculation used is seed inoculation, whether performed immediately before sowing or using preinoculated seeds or pretreated seeds by the professional seed treatment. The methodology of inoculation should not only cover the seeds with living rhizobia, but must also optimize the chances of these rhizobia to infect the roots and nodulate. To this end, inoculated rhizobia must be in such an amount and condition that would allow them to overcome the competition exerted by the rhizobia of the allochthonous population of the soil, which are usually less effective for nitrogen fixation and thus dilute the effect of inoculation on yield. This optimization requires solving some queries related to the current knowledge of seed inoculation, which are addressed in this article. I conclude that the aspects that require further research are the adhesion and survival of rhizobia on seeds, the release of rhizobia once the seeds are deposited in the soil, and the movement of rhizobia from the vicinity of the seeds to the infection sites in the roots. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  11. Sensibilidade de estirpes de Bradyrhizobium ao glyphosate

    Directory of Open Access Journals (Sweden)

    Rodrigo Josemar Seminoti Jacques

    2010-02-01

    Full Text Available A aplicação do glyphosate sobre a soja resistente a este herbicida pode causar prejuízos à simbiose com o rizóbio. O objetivo deste trabalho foi avaliar a sensibilidade ao herbicida glyphosate de três estirpes de Bradyrhizobium recomendadas para a produção de inoculantes de sementes de soja no Brasil. Avaliou-se o efeito das concentrações de 0,0; 5,4; 10,8; 21,6 e 43,2 µg L-1 do ingrediente ativo do glyphosate [N-(fosfonometil glicina] no meio YM líquido sobre o crescimento de B. japonicum (estirpe SEMIA 5079 e de B. elkanii (estirpe SEMIA 5019 e estirpe SEMIA 587, por meio de leituras das densidades óticas e geração de curvas de crescimento. As reduções de crescimento na presença da menor concentração do glyphosate foram de 18% para SEMIA 5079, 29% para SEMIA 5019 e de 35% para SEMIA 587, sendo, de modo geral, quanto maior a concentração do herbicida no meio de cultura maior a inibição do crescimen­to. As estirpes apresentaram sensibilidade diferencial somente às concentrações mais baixas do glyphosate; nesse caso, foi possível determinar a seguinte ordem de sensibilidade: SEMIA 587 > SEMIA 5019 > SEMIA 5079. Essa sensibilidade diferencial é dependente da concentração do herbicida, pois na presença de 43,2 µg L-1 todas as estirpes tiveram seu crescimento severamente reduzido, não havendo diferença entre elas.

  12. Taxonomy Icon Data: Schistosoma japonicum [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Schistosoma japonicum Schistosoma japonicum Platyhelminthes Schistosoma_japonicum_L.png Schistosoma_japon...icum_NL.png Schistosoma_japonicum_S.png Schistosoma_japonicum_NS.png http://bioscience...dbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japonicum&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japon...icum&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japon...icum&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japonicum&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=132 ...

  13. Phylogeny of nodulation and nitrogen-fixation genes in Bradyrhizobium: supporting evidence for the theory of monophyletic origin, and spread and maintenance by both horizontal and vertical transfer.

    Science.gov (United States)

    Menna, Pâmela; Hungria, Mariangela

    2011-12-01

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

  14. Strains nodulating Lupinus albus on different continents belong to several new chromosomal and symbiotic lineages within Bradyrhizobium.

    Science.gov (United States)

    Velázquez, Encarna; Valverde, Angel; Rivas, Raúl; Gomis, Víctor; Peix, Alvaro; Gantois, Inne; Igual, José M; León-Barrios, Milagros; Willems, Anne; Mateos, Pedro F; Martínez-Molina, Eustoquio

    2010-05-01

    In this work we analysed different chromosomal and symbiotic markers in rhizobial strains nodulating Lupinus albus (white lupin) in several continents. Collectively the analysis of their rrs and atpD genes, and 16S-23S intergenic spacers (ITS), showed that they belong to at least four chromosomal lineages within the genus Bradyrhizobium. Most isolates from the Canary Islands (near to the African continent) grouped with some strains isolated on mainland Spain and were identified as Bradyrhizobium canariense. These strains are divided into two ITS subgroups coincident with those previously described from isolates nodulating Ornithopus. The remaining strains isolated on mainland Spain grouped with most isolates from Chile (American continent) forming a new lineage related to Bradyrhizobium japonicum. The strains BLUT2 and ISLU207 isolated from the Canary Islands and Chile, respectively, formed two new lineages phylogenetically close to different species of Bradyrhizobium depending on the marker analyzed. The analysis of the nodC gene showed that all strains nodulating L. albus belong to the biovar genistearum; nevertheless they form four different nodC lineages of which lineage C is at present exclusively formed by L. albus endosymbionts isolated from different continents.

  15. Microevolution of symbiotic Bradyrhizobium populations associated with soybeans in east North America

    Science.gov (United States)

    Tang, Jie; Bromfield, E S P; Rodrigue, N; Cloutier, S; Tambong, J T

    2012-01-01

    Microevolution and origins of Bradyrhizobium populations associated with soybeans at two field sites (A and B, 280 km apart in Canada) with contrasting histories of inoculation was investigated using probabilistic analyses of six core (housekeeping) gene sequences. These analyses supported division of 220 isolates in five lineages corresponding either to B. japonicum groups 1 and 1a or to one of three novel lineages within the genus Bradyrhizobium. None of the isolates from site A and about 20% from site B (the only site with a recent inoculation history) were attributed to inoculation sources. The data suggest that most isolates were of indigenous origin based on sequence analysis of 148 isolates of soybean-nodulating bacteria from native legumes (Amphicarpaea bracteata and Desmodium canadense). Isolates from D. canadense clustered with B. japonicum group 1, whereas those from A. bracteata were placed in two novel lineages encountered at soybean field sites. One of these novel lineages predominated at soybean sites and exhibited a significant clonal expansion likely reflecting selection by the plant host. Homologous recombination events detected in the 35 sequence types from soybean sites had an effect on genetic diversification that was approximately equal to mutation. Interlineage transfer of core genes was infrequent and mostly attributable to gyrB that had a history of frequent recombination. Symbiotic gene sequences (nodC and nifH) of isolates from soybean sites and native legumes clustered in two lineages corresponding to B. japonicum and B. elkani with the inheritance of these genes appearing predominantly by vertical transmission. The data suggest that soybean-nodulating bacteria associated with native legumes represent a novel source of ecologically adapted bacteria for soybean inoculation. PMID:23301163

  16. Colony Dimorphism in Bradyrhizobium Strains

    Science.gov (United States)

    Sylvester-Bradley, Rosemary; Thornton, Philip; Jones, Peter

    1988-01-01

    Ten isolates of Bradyrhizobium spp. which form two colony types were studied; the isolates originated from a range of legume species. The two colony types differed in the amount of gum formed or size or both, depending on the strain. Whole 7-day-old colonies of each type were subcultured to determine the proportion of cells which had changed to the other type. An iterative computerized procedure was used to determine the rate of switching per generation between the two types and to predict proportions reached at equilibrium for each strain. The predicted proportions of the wetter (more gummy) or larger colony type at equilibrium differed significantly between strains, ranging from 0.9999 (strain CIAT 2383) to 0.0216 (strain CIAT 2469), because some strains switched faster from dry to wet (or small to large) and others switched faster from wet to dry (or large to small). Predicted equilibrium was reached after about 140 generations in strain USDA 76. In all but one strain (CIAT 3030) the growth rate of the wetter colony type was greater than or similar to that of the drier type. The mean difference in generation time between the two colony types was 0.37 h. Doubling times calculated for either colony type after 7 days of growth on the agar surface ranged from 6.0 to 7.3 h. The formation of two persistent colony types by one strain (clonal or colony dimorphism) may be a common phenomenon among Bradyrhizobium strains. Images PMID:16347599

  17. Bradyrhizobium strain and the {sup 15}N natural abundance quantification of biological N{sub 2} fixation in soybean

    Energy Technology Data Exchange (ETDEWEB)

    Guimaraes, Ana Paula [Universidade Estadual do Norte Fluminense (UENF), Campos dos Goytacases, RJ (Brazil). Centro de Ciencias e Tecnologias Agropecuarias (CCTA). Dept. de Producao Vegetal; Morais, Rafael Fiusa de; Urquiaga, Segundo; Boddey, Robert Michael; Alves, Bruno Jose Rodrigues [EMBRAPA Agrobiologia, Seropedica, RJ (Brazil)]. E-mail: bruno@cnpab.embrapa.br

    2008-09-15

    In commercial plantations of soybean in both the Southern and the Cerrado regions, contributions from biological nitrogen fixation (BNF) are generally proportionately high. When using the {sup 15}N natural abundance technique to quantify BNF inputs, it is essential to determine, with accuracy, the {sup 15}N abundance of the N derived from BNF (the 'B' value). This study aimed to determine the effect of four recommended strains of Bradyrhizobium spp. (two B. japonicum and two B. elkanii) on the 'B' value of soybean grown in pots in an open field using an equation based on the determination of {delta}{sup 15}N natural abundance in a non-labelled soil, and estimate of the contribution of BNF derived from the use of {sup 15}N-isotope dilution in soils enriched with {sup 15}N. To evaluate N{sub 2} fixation by soybean, three non-N{sub 2}-fixing reference crops were grown under the same conditions. Regardless of Bradyrhizobium strain, no differences were observed in dry matter, nodule weight and total N between labelled and non-labelled soil. The N{sub 2} fixation of the soybeans grown in the two soil conditions were similar. The mean 'B' values of the soybeans inoculated with the B. japonicum strains were -1.84 per mille and -0.50 per mille, while those inoculated with B. elkanii were -3.67 per mille and -1.0 per mille, for the shoot tissue and the whole plant, respectively. Finally, the 'B' value for the soybean crop varied considerably in function of the inoculated Bradyrhizobium strain, being most important when only the shoot tissue was utilised to estimate the proportion of N in the plant derived from N{sub 2} fixation. (author)

  18. The endemic Genista versicolor from Sierra Nevada National Park in Spain is nodulated by putative new Bradyrhizobium species and a novel symbiovar (sierranevadense).

    Science.gov (United States)

    Cobo-Díaz, José F; Martínez-Hidalgo, Pilar; Fernández-González, Antonio J; Martínez-Molina, Eustoquio; Toro, Nicolás; Velázquez, Encarna; Fernández-López, Manuel

    2014-05-01

    Genista versicolor is an endemic legume from Sierra Nevada National Park which constitutes one of the UNESCO-recognized Biosphere Reserves. In the present study, a collection of strains nodulating this legume was analysed in characteristic soils of this ecosystem. Most strains nodulating G. versicolor belonged to rrs group I within the genus Bradyrhizobium and only one strain, named GV137, belonged to rrs group II from which only a single species, B. retamae, has been described in Europe to date. Strain GV137, and some strains from rrs group I, belonged to putative new species of Bradyrhizobium, although most strains from group I belonged to B. canariense, according to the ITS fragment and atpD gene analysis. This result contrasted with those obtained in Genista tinctoria in Northeast Europe whose endosymbionts were identified as B. japonicum. The analysis of the symbiotic nodC and nifH genes carried by G. versicolor-nodulating strains showed that most of them belonged to symbiovar genistearum, as did those isolated from G. tinctoria. Nevertheless, strain GV137, belonging to rrs group II, formed a divergent lineage that constituted a novel symbiovar within the genus Bradyrhizobium for which the name sierranevadense is proposed. This finding showed that the Genisteae are not restrictive legumes only nodulated by symbiovar genistearum, since Genista is a promiscuous legume nodulated by at least two symbiovars of Bradyrhizobium, as occurs in Retama species. Copyright © 2013. Published by Elsevier GmbH.

  19. Isolation and selection of Bradyrhizobium from the root nodules of ...

    African Journals Online (AJOL)

    This research was conducted from 2004 to 2005 to isolate and select Bradyrhizobium from the root nodules of indigo plants (Indigofera tinctoria L.). For isolation of Bradyrhizobium, root nodules were collected from indigo plants. Fourteen Bradyrhizobium isolates were identified depending on colony, morphological and ...

  20. Endosymbiotic bacteria nodulating a new endemic lupine Lupinus mariae-josephi from alkaline soils in Eastern Spain represent a new lineage within the Bradyrhizobium genus.

    Science.gov (United States)

    Sánchez-Cañizares, Carmen; Rey, Luis; Durán, David; Temprano, Francisco; Sánchez-Jiménez, Paloma; Navarro, Albert; Polajnar, Mira; Imperial, Juan; Ruiz-Argüeso, Tomás

    2011-05-01

    Lupinus mariae-josephi is a recently described endemic Lupinus species from a small area in Eastern Spain where it thrives in soils with active lime and high pH. The L. mariae-josephi root symbionts were shown to be very slow-growing bacteria with different phenotypic and symbiotic characteristics from those of Bradyrhizobium strains nodulating other Lupinus. Their phylogenetic status was examined by multilocus sequence analyses of four housekeeping genes (16S rRNA, glnII, recA, and atpD) and showed the existence of a distinct evolutionary lineage for L. mariae-josephi that also included Bradyrhizobium jicamae. Within this lineage, the tested isolates clustered in three different sub-groups that might correspond to novel sister Bradyrhizobium species. These core gene analyses consistently showed that all the endosymbiotic bacteria isolated from other Lupinus species of the Iberian Peninsula were related to strains of the B. canariense or B. japonicum lineages and were separate from the L. mariae-josephi isolates. Phylogenetic analysis based on nodC symbiotic gene sequences showed that L. mariae-josephi bacteria also constituted a new symbiotic lineage distant from those previously defined in the genus Bradyrhizobium. In contrast, the nodC genes of isolates from other Lupinus spp. from the Iberian Peninsula were again clearly related to the B. canariense and B. japonicum bv. genistearum lineages. Speciation of L. mariae-josephi bradyrhizobia may result from the colonization of a singular habitat by their unique legume host. Copyright © 2011 Elsevier GmbH. All rights reserved.

  1. Efeitos de diferentes formulações comerciais de glyphosate sobre estirpes de Bradyrhizobium Effects of different glyphosate commercial formulations on Bradyrhizobium strains

    Directory of Open Access Journals (Sweden)

    J.B. Santos

    2004-06-01

    Full Text Available O objetivo deste trabalho foi avaliar efeitos de formulações comerciais de glyphosate sobre estirpes de Bradyrhizobium, em condições de laboratório. As formulações foram aplicadas na concentração de 43,2 µg L-1 do equivalente ácido. As bactérias foram inoculadas em meio de cultura à base de manitol e extrato de levedura (YM. O efeito do herbicida no crescimento das estirpes de Bradyrhizobium foi avaliado mediante leitura da densidade ótica em espectrofotômetro. Avaliou-se o crescimento das estirpes de B. japonicum SEMIA 5079 e de B. elkanii SEMIA 5019 e SEMIA 587 sob efeito de nove formulações de glyphosate: Zapp Qi®, Roundup®, Roundup Multiação®, Roundup Transorb®, Roundup WG®, Trop®, Agrisato®, glyphosate técnico [padrão de N-(phosphonomethyl glycina] e controle sem adição de herbicida (testemunha para as estirpes. Foram utilizadas seis repetições. Confeccionaram-se curvas de crescimento para cada estirpe. Pelos resultados, pôde-se observar que todas as formulações de glyphosate causaram efeitos diferenciados sobre as estirpes de Bradyrhizobium SEMIA 5019, SEMIA 5079 e SEMIA 587. Constatou-se que a formulação Zapp Qi foi a menos tóxica às estirpes de Bradyrhizobium avaliadas. A maior toxicidade foi observada para Roundup Transorb, que provocou reduções no crescimento acima de 94% para todas as estirpes de Bradyrhizobium estudadas. Não se observou correlação entre o tipo de sal - isopropilamina, amônio ou potássico, presentes na formulação herbicida - e o grau de inibição no crescimento das estirpes. SEMIA 587 foi a estirpe menos tolerante à maioria das formulações testadas, porém SEMIA 5019 foi a mais sensível ao glyphosate padrão, sem adição de sais ou de outros aditivos.This work aimed to evaluate the effects of glyphosate commercial formulations on Bradyrhizobium strains under laboratory conditions. The formulations were applied in the concentration of 43.2 µg L-1 of the a.e. and

  2. Pectolinarigenin - A Flavonoid Compound from Cirsium Japonicum ...

    African Journals Online (AJOL)

    HP

    the extract of C. japonicum and its major constituents possess anti-tumor [1, 2], anti- diabetic [3], antioxidant ... Extraction and isolation of pectolinarigenin. The powdered air-dried aerial parts of C. japonicum (1 kg) .... Caspase 3 is a member of the cysteine-aspartic acid protease family and a critical executioner of apoptosis ...

  3. Drivers of Symbiotic Quality in Wild Bradyrhizobium

    OpenAIRE

    Gano, Kelsey Annette

    2016-01-01

    Understanding the drivers of variation in symbiont quality is a fundamental objective in the study of mutualisms. Eukaryotic hosts express control traits that can selectively favor beneficial symbionts over ineffective genotypes, but bacterial symbionts range widely in beneficial quality. Evolutionary instability in symbiotic function and/or context dependency in the expression of symbiotic traits are predicted to contribute to this variation. The Acmispon-Bradyrhizobium mutualism is a model ...

  4. Bradyrhizobium neotropicale sp. nov., isolated from effective nodules of Centrolobium paraense

    OpenAIRE

    Zilli, Jerri E; Baraúna, Alexandre C; da Silva, Krisle; De Meyer, Sofie; Farias, Eliane NC; Kaminski, Paulo E; da Costa, Ismaele B; Ardley, Julie K; Willems, Anne; Camacho, Natalia N; Dourado, Fernanda dos S; O'Hara, Graham

    2014-01-01

    Root nodule bacteria were isolated from Centrolobium paraense Tul. grown in soils from the Amazon region, State of Roraima (Brazil). 16S rRNA gene sequence analysis of seven strains (BR 10247(T), BR 10296, BR 10297, BR 10298, BR 10299, BR 10300 and BR 10301) placed them in the genus Bradyrhizobium with the closest neighbours being the type strains of Bradyrhizobium paxllaeri (98.8 % similarity), Bradyrhizobium icense (98.8 %), Bradyrhizobium lablabi (98.7 %), Bradyrhizobium jicamae (98.6 %), ...

  5. Phylogenies of symbiotic genes of Bradyrhizobium symbionts of legumes of economic and environmental importance in Brazil support the definition of the new symbiovars pachyrhizi and sojae.

    Science.gov (United States)

    Delamuta, Jakeline Renata Marçon; Menna, Pâmela; Ribeiro, Renan Augusto; Hungria, Mariangela

    2017-07-01

    Bradyrhizobium comprises most tropical symbiotic nitrogen-fixing strains, but the correlation between symbiotic and core genes with host specificity is still unclear. In this study, the phylogenies of the nodY/K and nifH genes of 45 Bradyrhizobium strains isolated from legumes of economic and environmental importance in Brazil (Arachis hypogaea, Acacia auriculiformis, Glycine max, Lespedeza striata, Lupinus albus, Stylosanthes sp. and Vigna unguiculata) were compared to 16S rRNA gene phylogeny and genetic diversity by rep-PCR. In the 16S rRNA tree, strains were distributed into two superclades-B. japonicum and B. elkanii-with several strains being very similar within each clade. The rep-PCR analysis also revealed high intra-species diversity. Clustering of strains in the nodY/K and nifH trees was identical: 39 strains isolated from soybean grouped with Bradyrhizobium type species symbionts of soybean, whereas five others occupied isolated positions. Only one strain isolated from Stylosanthes sp. showed similar nodY/K and nifH sequences to soybean strains, and it also nodulated soybean. Twenty-one representative strains of the 16S rRNA phylogram were selected and taxonomically classified using a concatenated glnII-recA phylogeny; nodC sequences were also compared and revealed the same clusters as observed in the nodY/K and nifH phylograms. The analyses of symbiotic genes indicated that a large group of strains from the B. elkanii superclade comprised the novel symbiovar sojae, whereas for another group, including B. pachyrhizi, the symbiovar pachyrhizi could be proposed. Other potential new symbiovars were also detected. The co-evolution hypotheses is discussed and it is suggested that nodY/K analysis would be useful for investigating the symbiotic diversity of the genus Bradyrhizobium. Copyright © 2017 Elsevier GmbH. All rights reserved.

  6. Non-symbiotic Bradyrhizobium ecotypes dominate North American forest soils

    National Research Council Canada - National Science Library

    VanInsberghe, David; Maas, Kendra R; Cardenas, Erick; Strachan, Cameron R; Hallam, Steven J; Mohn, William W

    2015-01-01

    The genus Bradyrhizobium has served as a model system for studying host-microbe symbiotic interactions and nitrogen fixation due to its importance in agricultural productivity and global nitrogen cycling...

  7. Occurrence of H2-Uptake Hydrogenases in Bradyrhizobium sp. (Lupinus) and Their Expression in Nodules of Lupinus spp. and Ornithopus compressus1

    Science.gov (United States)

    Murillo, Jesús; Villa, Ana; Chamber, Manuel; Ruiz-Argüeso, Tomás

    1989-01-01

    Fifty-four strains of Bradyrhizobium sp. (Lupinus) from worldwide collections were screened by a colony hybridization method for the presence of DNA sequences homologous to the structural genes of the Bradyrhizobium japonicum hydrogenase. Twelve strains exhibited strong colony hybridization signals, and subsequent Southern blot hybridization experiments showed that they fell into two different groups on the basis of the pattern of EcoRI fragments containing the homology to the hup probe. All strains in the first group (UPM860, UPM861, and 750) expressed uptake hydrogenase activity in symbiosis with Lupinus albus, Lupinus angustifolius, Lupinus luteus, and Ornithopus compressus, but both the rate of H2 uptake by bacteroids and the relative efficiency of N2 fixation (RE = 1 - [H2 evolved in air/acetylene reduced]) by nodules were markedly affected by the legume host. L. angustifolius was the less permissive host for hydrogenase expression in symbiosis with the three strains (average RE = 0.76), and O. compressus was the more permissive (average RE = 1.0). None of the strains in the second group expressed hydrogenase activity in lupine nodules, and only one exhibited low H2-uptake activity in symbiosis with O. compressus. The inability of these putative Hup+ strains to induce hydrogenase activity in lupine nodules is discussed on the basis of the legume host effect. Among the 42 strains showing no homology to the B. japonicum hup-specific probe in the colony hybridization assay, 10 were examined in symbiosis with L. angustifolius. The average RE for these strains was 0.51. However, one strain, IM43B, exhibited high RE values (higher than 0.80) and high levels of hydrogenase activity in symbiosis with L. angustifolius, L. albus, and L. luteus. In Southern blot hybridization experiments, no homology was detected between the B. japonicum hup-specific DNA probe and total DNA from vegetative cells or bacteroids from strain IM43B even under low stringency hybridization

  8. Diversity of Bradyrhizobium strains nodulating Lupinus micranthus on both sides of the Western Mediterranean: Algeria and Spain.

    Science.gov (United States)

    Bourebaba, Yasmina; Durán, David; Boulila, Farida; Ahnia, Hadjira; Boulila, Abdelghani; Temprano, Francisco; Palacios, José M; Imperial, Juan; Ruiz-Argüeso, Tomás; Rey, Luis

    2016-06-01

    Lupinus micranthus is a lupine distributed in the Mediterranean basin whose nitrogen fixing symbiosis has not been described in detail. In this study, 101 slow-growing nodule isolates were obtained from L. micranthus thriving in soils on both sides of the Western Mediterranean. The diversity of the isolates, 60 from Algeria and 41 from Spain, was addressed by multilocus sequence analysis of housekeeping genes (16S rRNA, atpD, glnII and recA) and one symbiotic gene (nodC). Using genomic fingerprints from BOX elements, 37 different profiles were obtained (22 from Algeria and 15 from Spain). Phylogenetic analysis based on 16S rRNA and concatenated atpD, glnII and recA sequences of a representative isolate of each BOX profile displayed a homogeneous distribution of profiles in six different phylogenetic clusters. All isolates were taxonomically ascribed to the genus Bradyrhizobium. Three clusters comprising 24, 6, and 4 isolates, respectively, accounted for most of the profiles. The largest cluster was close to the Bradyrhizobium canariense lineage, while the other two were related to B. cytisi/B. rifense. The three remaining clusters included only one isolate each, and were close to B. canariense, B. japonicum and B. elkanii species, respectively. In contrast, phylogenetic clustering of BOX profiles based on nodC sequences yielded only two phylogenetic groups. One of them included all the profiles except one, and belonged to symbiovar genistearum. The remaining profile, constituted by a strain related to B. elkanii, was not related to any well-defined symbiotic lineage, and may constitute both a new symbiovar and a new genospecies. Copyright © 2016 Elsevier GmbH. All rights reserved.

  9. Genetic diversity of symbiotic Bradyrhizobium elkanii populations recovered from inoculated and non-inoculated Acacia mangium field trials in Brazil.

    Science.gov (United States)

    Perrineau, M M; Le Roux, C; de Faria, S M; de Carvalho Balieiro, F; Galiana, A; Prin, Y; Béna, G

    2011-07-01

    Acacia mangium is a legume tree native to Australasia. Since the eighties, it has been introduced into many tropical countries, especially in a context of industrial plantations. Many field trials have been set up to test the effects of controlled inoculation with selected symbiotic bacteria versus natural colonization with indigenous strains. In the introduction areas, A. mangium trees spontaneously nodulate with local and often ineffective bacteria. When inoculated, the persistence of inoculants and possible genetic recombination with local strains remain to be explored. The aim of this study was to describe the genetic diversity of bacteria spontaneously nodulating A. mangium in Brazil and to evaluate the persistence of selected strains used as inoculants. Three different sites, several hundred kilometers apart, were studied, with inoculated and non-inoculated plots in two of them. Seventy-nine strains were isolated from nodules and sequenced on three housekeeping genes (glnII, dnaK and recA) and one symbiotic gene (nodA). All but one of the strains belonged to the Bradyrhizobium elkanii species. A single case of housekeeping gene transfer was detected among the 79 strains, suggesting an extremely low rate of recombination within B. elkanii, whereas the nodulation gene nodA was found to be frequently transferred. The fate of the inoculant strains varied depending on the site, with a complete disappearance in one case, and persistence in another. We compared our results with the sister species Bradyrhizobium japonicum, both in terms of population genetics and inoculant strain destiny. Copyright © 2011 Elsevier GmbH. All rights reserved.

  10. Genome analysis of a novel Bradyrhizobium sp. DOA9 carrying a symbiotic plasmid.

    Directory of Open Access Journals (Sweden)

    Shin Okazaki

    Full Text Available Bradyrhizobium sp. DOA9 isolated from the legume Aeschynomene americana exhibited a broad host range and divergent nodulation (nod genes compared with other members of the Bradyrhizobiaceae. Genome analysis of DOA9 revealed that its genome comprised a single chromosome of 7.1 Mbp and a plasmid of 0.7 Mbp. The chromosome showed highest similarity with that of the nod gene-harboring soybean symbiont B. japonicum USDA110, whereas the plasmid showed highest similarity with pBBta01 of the nod gene-lacking photosynthetic strain BTAi1, which nodulates Aeschynomene species. Unlike in other bradyrhizobia, the plasmid of DOA9 encodes genes related to symbiotic functions including nodulation, nitrogen fixation, and type III/IV protein secretion systems. The plasmid has also a lower GC content (60.1% than the chromosome (64.4%. These features suggest that the plasmid could be the origin of the symbiosis island that is found in the genome of other bradyrhizobia. The nod genes of DOA9 exhibited low similarity with those of other strains. The nif gene cluster of DOA9 showed greatest similarity to those of photosynthetic bradyrhizobia. The type III/IV protein secretion systems of DOA9 are similar to those of nod gene-harboring B. elkanii and photosynthetic BTAi1. The DOA9 genome exhibited intermediate characteristics between nod gene-harboring bradyrhizobia and nod gene-lacking photosynthetic bradyrhizobia, thus providing the evidence for the evolution of the Bradyrhizobiaceae during ecological adaptation. Bradyrhizobium sp. DOA9 isolated from the legume Aeschynomene americana exhibited a broad host range and divergent nodulation (nod genes compared with other members of the Bradyrhizobiaceae. Genome analysis of DOA9 revealed that its genome comprised a single chromosome of 7.1 Mbp and a plasmid of 0.7 Mbp. The chromosome showed highest similarity with that of the nod gene-harboring soybean symbiont B. japonicum USDA110, whereas the plasmid showed highest

  11. Bradyrhizobium tropiciagri sp. nov. and Bradyrhizobium embrapense sp. nov., nitrogen-fixing symbionts of tropical forage legumes.

    Science.gov (United States)

    Delamuta, Jakeline Renata Marçon; Ribeiro, Renan Augusto; Ormeño-Orrillo, Ernesto; Parma, Marcia Maria; Melo, Itamar Soares; Martínez-Romero, Esperanza; Hungria, Mariangela

    2015-12-01

    Biological nitrogen fixation is a key process for agricultural production and environmental sustainability, but there are comparatively few studies of symbionts of tropical pasture legumes, as well as few described species of the genus Bradyrhizobium, although it is the predominant rhizobial genus in the tropics. A detailed polyphasic study was conducted with two strains of the genus Bradyrhizobium used in commercial inoculants for tropical pastures in Brazil, CNPSo 1112T, isolated from perennial soybean (Neonotonia wightii), and CNPSo 2833T, from desmodium (Desmodium heterocarpon). Based on 16S-rRNA gene phylogeny, both strains were grouped in the Bradyrhizobium elkanii superclade, but were not clearly clustered with any known species. Multilocus sequence analysis of three (glnII, gyrB and recA) and five (plus atpD and dnaK) housekeeping genes confirmed that the strains are positioned in two distinct clades. Comparison with intergenic transcribed spacer sequences of type strains of described species of the genus Bradyrhizobium showed similarity lower than 93.1 %, and differences were confirmed by BOX-PCR analysis. Nucleotide identity of three housekeeping genes with type strains of described species ranged from 88.1 to 96.2 %. Average nucleotide identity of genome sequences showed values below the threshold for distinct species of the genus Bradyrhizobium ( < 90.6 %), and the value between the two strains was also below this threshold (91.2 %). Analysis of nifH and nodC gene sequences positioned the two strains in a clade distinct from other species of the genus Bradyrhizobium. Morphophysiological, genotypic and genomic data supported the description of two novel species in the genus Bradyrhizobium, Bradyrhizobium tropiciagri sp. nov. (type strain CNPSo 1112T = SMS 303T = BR 1009T = SEMIA 6148T = LMG 28867T) and Bradyrhizobium embrapense sp. nov. (type strain CNPSo 2833T = CIAT 2372T = BR 2212T = SEMIA 6208T = U674T

  12. A new serratene triterpenoid from Lycopodium japonicum.

    Science.gov (United States)

    Sun, Zhang-Hua; Li, Wei; Tang, Gui-Hua; Yin, Sheng

    2017-03-01

    Phytochemical investigation on the herbs of Lycopodium japonicum led to the isolation of a new serratene triterpenoid, 3α,21α-dihydroxy-16-oxoserrat-14-en-24-yl p-coumarate (1), together with two known ones, lycernuic ketone C (2) and tohogenol (3). Their structures were elucidated by extensive spectroscopic methods, including 1D- and 2D-NMR and HR-ESI-MS. The 13 C NMR data of tohogenol was first reported.

  13. Symbiotic properties of Bradyrhizobium sp. (Lupinus assayed on serradella plants

    Directory of Open Access Journals (Sweden)

    Mieczysława Deryło

    2014-01-01

    Full Text Available Physiological and symbiotic properties of Bradyrhizobium sp. (Lupinus nodule isolates were compared to the standard slow-growing Bradyrhizobium sp. (Lupinus strain USDA 3045. Lupine nodules isolates showed typical characteristics for bradyrhizobial strains and nodulated small seed legume, serradella (Ornithopus sativus, in tube test. We observed a permanent physiological segregation of the effective (Fix' and ineffective (Fix- symbiotic phenotype for all tested bradyrhizobial strains during the growth of serradella in plant tube test. The ultrastructural differences between Fix* and Fix serradella nodules were observed. Rapid and visible nodulation as well as easy assay of the reduction of acetylene make serradella a convenient system for studies of Bradyrhizobium sp. (Lupinus strains in laboratory conditions.

  14. Photosynthetic adaptation of soybean due to varying effectiveness of N2 fixation by two distinct Bradyrhizobium japonicum strains

    NARCIS (Netherlands)

    Kaschuk, G.; Yin, X.; Hungria, M.; Leffelaar, P.A.; Giller, K.E.; Kuyper, T.W.

    2012-01-01

    Rhizobial N2 fixation is a costly biochemical process, which takes 6–14% of current photosynthate (C) from legumes, without compromising grain productivity. In addition to the effects of leaf N nutrition, rhizobial symbiosis could stimulate photosynthesis due to the removal of C sink limitation by

  15. Functional characterisation of Schistosoma japonicum acetylcholinesterase.

    Science.gov (United States)

    You, Hong; Gobert, Geoffrey N; Du, Xiaofeng; Pali, Gabor; Cai, Pengfei; Jones, Malcolm K; McManus, Donald P

    2016-06-10

    Acetylcholinesterase (AChE) is an important metabolic enzyme of schistosomes present in the musculature and on the surface of the blood stage where it has been implicated in the modulation of glucose scavenging from mammalian host blood. As both a target for the antischistosomal drug metrifonate and as a potential vaccine candidate, AChE has been characterised in the schistosome species Schistosoma mansoni, S. haematobium and S. bovis, but not in S. japonicum. Recently, using a schistosome protein microarray, a predicted S. japonicum acetylcholinesterase precursor was significantly targeted by protective IgG1 immune responses in S. haematobium-exposed individuals that had acquired drug-induced resistance to schistosomiasis after praziquantel treatment. We report the full-length cDNA sequence and describe phylogenetic and molecular structural analysis to facilitate understanding of the biological function of AChE (SjAChE) in S. japonicum. The protein has high sequence identity (88 %) with the AChEs in S. mansoni, S. haematobium and S. bovis and has 25 % sequence similarity with human AChE, suggestive of a highly specialised role for the enzyme in both parasite and host. We immunolocalized SjAChE and demonstrated its presence on the surface of adult worms and schistosomula, as well as its lower expression in parenchymal regions. The relatively abundance of AChE activity (90 %) present on the surface of adult S. japonicum when compared with that reported in other schistosomes suggests SjAChE may be a more effective drug or immunological target against this species. We also demonstrate that the classical inhibitor of AChE, BW285c51, inhibited AChE activity in tegumental extracts of paired worms, single males and single females by 59, 22 and 50 %, respectively, after 24 h incubation with 200 μM BW284c51. These results build on previous studies in other schistosome species indicating major differences in the enzyme between parasite and mammalian host, and provide

  16. Bradyrhizobium centrolobii and Bradyrhizobium macuxiense sp. nov. isolated from Centrolobium paraense grown in soil of Amazonia, Brazil.

    Science.gov (United States)

    Michel, Daniele C; Passos, Samuel R; Simões-Araujo, Jean L; Baraúna, Alexandre C; da Silva, Krisle; Parma, Marcia M; Melo, Itamar S; De Meyer, Sofie E; O'Hara, Graham; Zilli, Jerri E

    2017-07-01

    Thirteen Gram-negative, aerobic, motile with polar flagella, rod-shaped bacteria were isolated from root nodules of Centrolobium paraense Tul. grown in soils from the Amazon region of Brazil. Growth of strains was observed at temperature range 20-36 °C (optimal 28 °C), pH ranges 5-11 (optimal 6.0-7.0), and 0.1-0.5%NaCl (optimal 0.1-0.3%). Analysis of 16S rRNA gene placed the strains into two groups within Bradyrhizobium. Closest neighbouring species (98.8%) for group I was B. neotropicale while for group II were 12 species with more than 99% of similarity. Multi-locus sequence analysis (MLSA) with dnaK, glnII, recA, and rpoB confirmed B. neotropicale BR 10247 T as the closest type strain for the group I and B. elkanii USDA 76 T and B. pachyrhizi PAC 48 T for group II. Average Nucleotide Identity (ANI) differentiated group I from the B. neotropicale BR 10247 T (79.6%) and group II from B. elkanii USDA 76 T and B. pachyrhizi PAC 48 T (88.1% and 87.9%, respectively). Fatty acid profiles [majority C 16:0 and Summed feature 8 (18:1ω6c/18:1ω7c) for both groups], DNA G + C content, and carbon compound utilization supported the placement of the novel strains in the genus Bradyrhizobium. Gene nodC and nifH of the new strains have in general low similarity with other Bradyrhizobium species. Both groups nodulated plants from the tribes Crotalarieae, Dalbergiae, Genisteae, and Phaseoleae. Based on the presented data, two novel species which the names Bradyrhizobium centrolobii and Bradyrhizobium macuxiense are proposed, with BR 10245 T (=HAMBI 3597 T ) and BR 10303 T (=HAMBI 3602 T ) as the respective-type strains.

  17. Effects of genotype x bradyrhizobium inoculation or x fertilizer n ...

    African Journals Online (AJOL)

    symbiotic fixation). A set of 52 random lines from a broad-based reference population was evaluated for 2 years with and without B.japonicum inoculation in a split-plot design. Inoculation increased ... Genotype x: Inoculation Interactions, Nitrogen fixation ..... genotype x N treatment interaction was non significant each year ...

  18. Non-symbiotic Bradyrhizobium ecotypes dominate North American forest soils.

    Science.gov (United States)

    VanInsberghe, David; Maas, Kendra R; Cardenas, Erick; Strachan, Cameron R; Hallam, Steven J; Mohn, William W

    2015-11-01

    The genus Bradyrhizobium has served as a model system for studying host-microbe symbiotic interactions and nitrogen fixation due to its importance in agricultural productivity and global nitrogen cycling. In this study, we identify a bacterial group affiliated with this genus that dominates the microbial communities of coniferous forest soils from six distinct ecozones across North America. Representative isolates from this group were obtained and characterized. Using quantitative population genomics, we show that forest soil populations of Bradyrhizobium represent ecotypes incapable of nodulating legume root hairs or fixing atmospheric nitrogen. Instead, these populations appear to be free living and have a greater potential for metabolizing aromatic carbon sources than their close symbiotic relatives. In addition, we identify fine-scaled differentiation between populations inhabiting neighboring soil layers that illustrate how diversity within Bradyrhizobium is structured by habitat similarity. These findings reconcile incongruent observations about this widely studied and important group of bacteria and highlight the value of ecological context to interpretations of microbial diversity and taxonomy. These results further suggest that the influence of this genus likely extends well beyond facilitating agriculture, especially as forest ecosystems are large and integral components of the biosphere. In addition, this study demonstrates how focusing research on economically important microorganisms can bias our understanding of the natural world.

  19. Bradyrhizobium elkanii nod regulon: insights through genomic analysis

    Directory of Open Access Journals (Sweden)

    Luciane M. P. Passaglia

    2017-07-01

    Full Text Available Abstract A successful symbiotic relationship between soybean [Glycine max (L. Merr.] and Bradyrhizobium species requires expression of the bacterial structural nod genes that encode for the synthesis of lipochitooligosaccharide nodulation signal molecules, known as Nod factors (NFs. Bradyrhizobium diazoefficiens USDA 110 possesses a wide nodulation gene repertoire that allows NF assembly and modification, with transcription of the nodYABCSUIJnolMNOnodZ operon depending upon specific activators, i.e., products of regulatory nod genes that are responsive to signaling molecules such as flavonoid compounds exuded by host plant roots. Central to this regulatory circuit of nod gene expression are NodD proteins, members of the LysR-type regulator family. In this study, publicly available Bradyrhizobium elkanii sequenced genomes were compared with the closely related B. diazoefficiens USDA 110 reference genome to determine the similarities between those genomes, especially with regards to the nod operon and nod regulon. Bioinformatics analyses revealed a correlation between functional mechanisms and key elements that play an essential role in the regulation of nod gene expression. These analyses also revealed new genomic features that had not been clearly explored before, some of which were unique for some B. elkanii genomes.

  20. [Identification of glycosylphosphatidylinositol-anchored protein from Schistosoma japonicum].

    Science.gov (United States)

    Cao, Qin-Yan; Xue, Yan-Feng; Shen, Li

    2012-10-30

    To identify glycosylphosphatidylinositol (GPI) anchored protein of Schistosoma japonicum. Based on the gene sequence of Schistosoma mansoni GPI anchored protein Sm200 (GenBank Assess No: XM_002569560.1), bioinformatics analysis was performed to find out its homologous gene sequence in S. japonicum, then a selected partial coding sequence (SjGPIs, about 933 bp) from the homologous gene sequence were amplified, and cloned into PET-28a(+) vector. The recombinant plasmid pET-28a(+)SjGPIs were transformed into E. coli Top10 cells and induced with IPTG for protein expression. The recombinant protein SjGPIs was purified with Ni-NTA resin, and the purified recombinant SjGPIs protein was used as antigen to prepare antiserum in New Zealand rabbit. The antiserum was used to detect S. japonicum GPI-anchored protein. To identify a GPI-anchored protein, the detected protein were identified by phosphatidylinositol-specific phospholipase C (PI-PLC) digestion. White blood cells from S. japonicum-infected mice was examined whether they endocytosed GPI-anchored proteins by Western blotting. The homologous gene sequence of S. mansoni GPI Sm200 gene was found in S. japonicum genome. A 3 495 bp coding sequence was obtained, containing the complete C-terminal sequence. The selected gene sequence (SjGPIs) were amplified and the recombinant plasmid pET-28a(+)-SjGPIs was established. According to the analysis of C-terminal sequence, Western blotting and enzyme digestion of PI-PLC, a GPI-anchored protein was present in S. japonicum tegument (about 1M(r)200000), named SjGPI200. The protein was detected in white blood cells of infected mice. SjGPI200 protein exists in S. japonicum, and anchored to parasite tegument via GPI.

  1. Bradyrhizobium neotropicale sp. nov., isolated from effective nodules of Centrolobium paraense.

    Science.gov (United States)

    Zilli, Jerri E; Baraúna, Alexandre C; da Silva, Krisle; De Meyer, Sofie E; Farias, Eliane N C; Kaminski, Paulo E; da Costa, Ismaele B; Ardley, Julie K; Willems, Anne; Camacho, Natália N; Dourado, Fernanda dos S; O'Hara, Graham

    2014-12-01

    Root nodule bacteria were isolated from Centrolobium paraense Tul. grown in soils from the Amazon region, State of Roraima (Brazil). 16S rRNA gene sequence analysis of seven strains (BR 10247(T), BR 10296, BR 10297, BR 10298, BR 10299, BR 10300 and BR 10301) placed them in the genus Bradyrhizobium with the closest neighbours being the type strains of Bradyrhizobium paxllaeri (98.8 % similarity), Bradyrhizobium icense (98.8 %), Bradyrhizobium lablabi (98.7 %), Bradyrhizobium jicamae (98.6 %), Bradyrhizobium elkanii (98.6 %), Bradyrhizobium pachyrhizi (98.6 %) and Bradyrhizobium retamae (98.3 %). This high similarity, however, was not confirmed by the intergenic transcribed spacer (ITS) 16S-23S rRNA region sequence analysis nor by multi-locus sequence analysis. Phylogenetic analyses of five housekeeping genes (dnaK, glnII, gyrB, recA and rpoB) revealed Bradyrhizobium iriomotense EK05(T) ( = LMG 24129(T)) to be the most closely related type strain (95.7 % sequence similarity or less). Chemotaxonomic data, including fatty acid profiles [major components being C16 : 0 and summed feature 8 (18 : 1ω6c/18 : 1ω7c)], DNA G+C content, slow growth rate and carbon compound utilization patterns, supported the placement of the novel strains in the genus Bradyrhizobium. Results of DNA-DNA relatedness studies and physiological data (especially carbon source utilization) differentiated the strains from the closest recognized species of the genus Bradyrhizobium. Symbiosis-related genes for nodulation (nodC) and nitrogen fixation (nifH) placed the novel species in a new branch within the genus Bradyrhizobium. Based on the current data, these seven strains represent a novel species for which the name Bradyrhizobium neotropicale sp. nov. is proposed. The type strain is BR 10247(T) ( = HAMBI 3599(T)). © 2014 IUMS.

  2. Exploring molecular variation in Schistosoma japonicum in China

    Science.gov (United States)

    Young, Neil D.; Chan, Kok-Gan; Korhonen, Pasi K.; Min Chong, Teik; Ee, Robson; Mohandas, Namitha; Koehler, Anson V.; Lim, Yan-Lue; Hofmann, Andreas; Jex, Aaron R.; Qian, Baozhen; Chilton, Neil B.; Gobert, Geoffrey N.; McManus, Donald P.; Tan, Patrick; Webster, Bonnie L.; Rollinson, David; Gasser, Robin B.

    2015-01-01

    Schistosomiasis is a neglected tropical disease that affects more than 200 million people worldwide. The main disease-causing agents, Schistosoma japonicum, S. mansoni and S. haematobium, are blood flukes that have complex life cycles involving a snail intermediate host. In Asia, S. japonicum causes hepatointestinal disease (schistosomiasis japonica) and is challenging to control due to a broad distribution of its snail hosts and range of animal reservoir hosts. In China, extensive efforts have been underway to control this parasite, but genetic variability in S. japonicum populations could represent an obstacle to eliminating schistosomiasis japonica. Although a draft genome sequence is available for S. japonicum, there has been no previous study of molecular variation in this parasite on a genome-wide scale. In this study, we conducted the first deep genomic exploration of seven S. japonicum populations from mainland China, constructed phylogenies using mitochondrial and nuclear genomic data sets, and established considerable variation between some of the populations in genes inferred to be linked to key cellular processes and/or pathogen-host interactions. Based on the findings from this study, we propose that verifying intraspecific conservation in vaccine or drug target candidates is an important first step toward developing effective vaccines and chemotherapies against schistosomiasis. PMID:26621075

  3. Non-equilibrium plasma prevention of Schistosoma japonicum transmission

    Science.gov (United States)

    Wang, Xing-Quan; Wang, Feng-Peng; Chen, Wei; Huang, Jun; Bazaka, Kateryna; Ostrikov, Kostya (Ken)

    2016-10-01

    Schistosoma japonicum is a widespread human and animal parasite that causes intestinal and hepatosplenic schistosomiasis linked to colon, liver and bladder cancers, and anemia. Estimated 230 million people are currently infected with Schistosoma spp, with 779 million people at risk of contracting the parasite. Infection occurs when a host comes into contact with cercariae, a planktonic larval stage of the parasite, and can be prevented by inactivating the larvae, commonly by chemical treatment. We investigated the use of physical non-equilibrium plasma generated at atmospheric pressure using custom-made dielectric barrier discharge reactor to kill S. japonicum cercariae. Survival rate decreased with treatment time and applied power. Plasmas generated in O2 and air gas discharges were more effective in killing S. japonicum cercariae than that generated in He, which is directly related to the mechanism by which cercariae are inactivated. Reactive oxygen species, such as O atoms, abundant in O2 plasma and NO in air plasma play a major role in killing of S. japonicum cercariae via oxidation mechanisms. Similar level of efficacy is also shown for a gliding arc discharge plasma jet generated in ambient air, a system that may be more appropriate for scale-up and integration into existing water treatment processes.

  4. [Content of rare earth elements in wild Hypericum japonicum Thunb].

    Science.gov (United States)

    Wei, Zhen-Lin; Rui, Yu-Kui; Tian, Zhi-Huan

    2009-06-01

    Rare earth elements are important nutritional elements for human health, and today more and more attention has been paid to the effective components in Chinese traditional medicine, especially to rare earth elements. Fifteen rare earth elements in wild hypericum japonicum Thunb were analyzed by the methods of ICP-MS. The results showed that the concentrations of La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Yb, Tm, Lu and Y ranged from 6 ng x g(-1) x DW to 14 522 ng x g(-1) x DW, and among them the concentrations of La, Ce and Nd were higher than 2 000 ng x g(-1) x DW. Compared with the concentration of rare earth elements in rice, corn, wheat and barley, the total concentration of rare earth elements in hypericum japonicum Thunb was much higher, which could be the mechanism of curative effect of hypericum japonicum Thunb on liverish diseases. The character of elements and the content of rare earth elements in soil should be responsible for the difference, but the distributive mechanism of rare earth elements in hypericum japonicum Thunb should be further studied.

  5. Origin and Evolution of Nitrogen Fixation Genes on Symbiosis Islands and Plasmid in Bradyrhizobium

    OpenAIRE

    Okubo, Takashi; Piromyou, Pongdet; Tittabutr, Panlada; Teaumroong, Neung; Minamisawa, Kiwamu

    2016-01-01

    The nitrogen fixation (nif) genes of nodule-forming Bradyrhizobium strains are generally located on symbiosis islands or symbiosis plasmids, suggesting that these genes have been transferred laterally. The nif genes of rhizobial and non-rhizobial Bradyrhizobium strains were compared in order to infer the evolutionary histories of nif genes. Based on all codon positions, the phylogenetic tree of concatenated nifD and nifK sequences showed that nifDK on symbiosis islands formed a different clad...

  6. In vitro cultivation of Schistosoma japonicum-parasites and cells.

    Science.gov (United States)

    Ye, Qing; Dong, Hui-Fen; Grevelding, Christoph G; Hu, Min

    2013-12-01

    Schistosomiasis is a serious parasitic zoonosis caused by blood-dwelling flukes of the genus Schistosoma. Understanding functions of genes and proteins of this parasite is important for uncovering this pathogen's complex biology, which will provide valuable information to design new strategies for schistosomiasis control. Effective applications of molecular tools reported to investigate schistosome gene function, such as inhibitor studies and transgenesis, rely on the developments of in vitro cultivation system of this parasite and cells. Besides the in vitro culture studies dealing with Schistosoma mansoni, there are also numerous excellent studies about the in vitro cultivation of Schistosoma japonicum, which were performed by Chinese researchers and published in Chinese journals. Nearly every stage of the life-cycle of S. japonicum, including miracidia, mother sporocysts, cercariae, schistosomula, and egg-laying adult worms, was employed for developing in vitro cultivation methods, being accompanied by the introduction of several media and supplements that helped to improve culture conditions. It was not only possible to generate mother sporocysts from miracidia in vitro, but also to obtain adult worms from cercariae through in vitro cultivation. The main obstacles to complete the life cycle of S. japonicum in the lab are the transition from mother sporocysts to cercariae, and the production of fertilized and completely developed eggs by adult worms generated in vitro. With regard to cells from S. japonicum, besides established isolation protocols and morphological observations, media optimizations were conducted by using different chemical reagents, biological supplements and physical treatment. Among these, mutagens like N-methyl-N-nitro-N-nitrosoguanidine and the addition of extracellular matrix were found to be able to induce mitogenic activities. Although enzyme activities or the level of silver-stained nucleolar region associated protein in cultured cells

  7. Two New Bioactive α-Pyrones from Hypericum japonicum

    Directory of Open Access Journals (Sweden)

    Linzhen Hu

    2016-04-01

    Full Text Available Hypericum japonicum (Guttiferae, a type of annual or perennial herb, has been historically applied to cure infectious hepatitis, acute and chronic hepatitis, gastrointestinal disorder, and internal hemorrhage. In our successive studies on the genus Hypericum, two new α-pyrones termed japopyrones A and B (1 and 2 were isolated from H. japonicum. Their structures and absolute configurations were established by the comprehensive analyses of spectroscopic data, the application of the Single-crystal X-ray diffraction structural analysis, and the experimental electronic circular dichroism (ECD spectra. Bioactivity screenings suggested that compound 2 possessed the potential inhibition efficacy on lytic replication of Kaposi’s sarcoma associated herpesvirus (KSHV with an IC50 29.46 μM and a selective index of higher than 6.79, respectively.

  8. A New Lycopodine-type Alkaloid from Lycopodium japonicum.

    Science.gov (United States)

    Yang, Qian; Zhu, Yuquan; Peng, Wei; Zhan, Rui; Chen, Yegao

    2016-10-01

    A new lycopodine-type alkaloid, 12β-hydroxy-acetylfawcettiine N-oxide (1), together with seven known analogues, acetyllycoposerramine M (2), lycopodine (3), lycoclavine (4), diphaladine A (5), lycoposerramine K (6), 11β-hydroxy-12-epilycodoline (7) and fawcettiine (8), were isolated from Lycopodium japonicum. Their structures were established by mass spectrometry and 1D and 2D NMR techniques. The isolated alkaloids were assayed for their inhibition activities against acetylcholinesterase, but no inhibitory activities for the compounds were detected.

  9. Non-equilibrium plasma prevention of Schistosoma japonicum transmission

    OpenAIRE

    Xing-Quan Wang; Feng-Peng Wang; Wei Chen; Jun Huang; Kateryna Bazaka; Kostya (Ken) Ostrikov

    2016-01-01

    Schistosoma japonicum is a widespread human and animal parasite that causes intestinal and hepatosplenic schistosomiasis linked to colon, liver and bladder cancers, and anemia. Estimated 230 million people are currently infected with Schistosoma spp, with 779 million people at risk of contracting the parasite. Infection occurs when a host comes into contact with cercariae, a planktonic larval stage of the parasite, and can be prevented by inactivating the larvae, commonly by chemical treatmen...

  10. Hyperjapones A-E, Terpenoid Polymethylated Acylphloroglucinols from Hypericum japonicum.

    Science.gov (United States)

    Yang, Xing-Wei; Li, Yan-Ping; Su, Jia; Ma, Wei-Guang; Xu, Gang

    2016-04-15

    Hyperjapones A-E (1-5), novel terpenoid polymethylated acylphloroglucinols (TPAPs) with unusual architectures, were characterized from Hypericum japonicum. Their structures and absolute configurations were determined by comprehensive spectroscopic data and X-ray diffractions. Compound 1 was obtained as a racemic mixture and was separated by a column coated with cellulose tris(4-methylbenzoate) after attempts with various chiral materials. Compounds 1, 2, and 4 exhibited moderate antitumor activities in vitro.

  11. Salmonella typhimurium Infection Reduces Schistosoma japonicum Worm Burden in Mice

    OpenAIRE

    Xiaoyang Zhu; Lu Chen; Junfang Wu; Huiru Tang; Yulan Wang

    2017-01-01

    Coinfection of microorganisms is a common phenomenon in humans and animals. In order to further our understanding of the progress of coinfection and the possible interaction between different pathogens, we have built a coinfection mouse model with Schistosoma japonicum and Salmonella typhimurium, and used this model to investigate the systemic metabolic and immune responses using NMR-based metabonomics and immunological techniques. Our results show that Salmonella typhimurium (ATCC14028) infe...

  12. Detection of Extracellular Enzyme Activities in Ganoderma neo-japonicum

    OpenAIRE

    Jo, Woo-Sik; Park, Ha-Na; Cho, Doo-Hyun; Yoo, Young-Bok; Park, Seung-Chun

    2011-01-01

    The ability of Ganoderma to produce extracellular enzymes, including β-glucosidase, cellulase, avicelase, pectinase, xylanase, protease, amylase, and ligninase was tested in chromogenic media. β-glucosidase showed the highest activity, among the eight tested enzymes. In particular, Ganoderma neo-japonicum showed significantly stronger activity for β-glucosidase than that of the other enzymes. Two Ganoderma lucidum isolates showed moderate activity for avicelase; however, Ganoderma neo-japonic...

  13. A Review of Schistosomiasis in Indonesia with Reference to Schistosoma Japonicum

    OpenAIRE

    Liat, Lim Boo; M. Sudomo

    1987-01-01

    Tinjauan tentang Schistosoma di Indonesia ini mencakup Schistosoma japonicum, S. incognitum, S. spindale dan Trichohilharzia brevis. Tinjauan dibuat atas dasar laporan-laporan penelitian yang telah diterbitkan. Di dalamnya dapat dijumpai uraian singkat tentang S. spinale dan T. brevis. Dari banyak publikasi tentang S. japonicum dan S. incoganitum dapat disajikan uraian tentang peranan kedua parasit tersebut sebagai penyebab penyakit baik manusia maupun hewan.

  14. A REVIEW OF SCHISTOSOMIASIS IN INDONESIA WITH REFERENCE TO SCHISTOSOMA JAPONICUM

    OpenAIRE

    Lim Boo Liat; M. Sudomo

    2012-01-01

    Tinjauan tentang Schistosoma di Indonesia ini mencakup Schistosoma japonicum, S. incognitum, S. spindale dan Trichohilharzia brevis. Tinjauan dibuat atas dasar laporan-laporan penelitian yang telah diterbitkan. Di dalamnya dapat dijumpai uraian singkat tentang S. spinale dan T. brevis. Dari banyak publikasi tentang S. japonicum dan S. incoganitum dapat disajikan uraian tentang peranan kedua parasit tersebut sebagai penyebab penyakit baik manusia maupun hewan.

  15. Cytokine mRNA profiles in pigs exposed prenatally and postnatally to Schistosoma japonicum

    DEFF Research Database (Denmark)

    Techau, Michala E.; Johansen, Maria V.; Aasted, Bent

    2007-01-01

    The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed...

  16. A database for the taxonomic and phylogenetic identification of the genus Bradyrhizobium using multilocus sequence analysis.

    Science.gov (United States)

    Azevedo, Helton; Lopes, Fabricio; Silla, Paulo; Hungria, Mariangela

    2015-01-01

    Biological nitrogen fixation, with an emphasis on the legume-rhizobia symbiosis, is a key process for agriculture and the environment, allowing the replacement of nitrogen fertilizers, reducing water pollution by nitrate as well as emission of greenhouse gases. Soils contain numerous strains belonging to the bacterial genus Bradyrhizobium, which establish symbioses with a variety of legumes. However, due to the high conservation of Bradyrhizobium 16S rRNA genes - considered as the backbone of the taxonomy of prokaryotes - few species have been delineated. The multilocus sequence analysis (MLSA) methodology, which includes analysis of housekeeping genes, has been shown to be promising and powerful for defining bacterial species, and, in this study, it was applied to Bradyrhizobium, species, increasing our understanding of the diversity of nitrogen-fixing bacteria. Classification of bacteria of agronomic importance is relevant to biodiversity, as well as to biotechnological manipulation to improve agricultural productivity. We propose the construction of an online database that will provide information and tools using MLSA to improve phylogenetic and taxonomic characterization of Bradyrhizobium, allowing the comparison of genomic sequences with those of type and representative strains of each species. A database for the taxonomic and phylogenetic identification of the Bradyrhizobium, genus, using MLSA, will facilitate the use of biological data available through an intuitive web interface. Sequences stored in the on-line database can be compared with multiple sequences of other strains with simplicity and agility through multiple alignment algorithms and computational routines integrated into the database. The proposed database and software tools are available at http://mlsa.cnpso.embrapa.br, and can be used, free of charge, by researchers worldwide to classify Bradyrhizobium, strains; the database and software can be applied to replicate the experiments presented in

  17. Bradyrhizobium ottawaense sp. nov., a symbiotic nitrogen fixing bacterium from root nodules of soybeans in Canada

    Science.gov (United States)

    Yu, Xiumei; Cloutier, Sylvie; Tambong, James T.

    2014-01-01

    Sixteen strains of symbiotic bacteria from root nodules of Glycine max grown in Ottawa, Canada, were previously characterized and placed in a novel group within the genus Bradyrhizobium. To verify their taxonomic status, these strains were further characterized using a polyphasic approach. All strains possessed identical 16S rRNA gene sequences that were 99.79 % similar to the closest relative, Bradyrhizobium liaoningense LMG 18230T. Phylogenetic analysis of concatenated atpD, glnII, recA, gyrB, rpoB and dnaK genes divided the 16 strains into three multilocus sequence types that were placed in a highly supported lineage distinct from named species of the genus Bradyrhizobium consistent with results of DNA–DNA hybridization. Based on analysis of symbiosis gene sequences (nodC and nifH), all novel strains were placed in a phylogenetic group with five species of the genus Bradyrhizobium that nodulate soybeans. The combination of phenotypic characteristics from several tests including carbon and nitrogen source utilization and antibiotic resistance could be used to differentiate representative strains from recognized species of the genus Bradyrhizobium. Novel strain OO99T elicits effective nodules on Glycine max, Glycine soja and Macroptilium atropurpureum, partially effective nodules on Desmodium canadense and Vigna unguiculata, and ineffective nodules on Amphicarpaea bracteata and Phaseolus vulgaris. Based on the data presented, we conclude that our strains represent a novel species for which the name Bradyrhizobium ottawaense sp. nov. is proposed, with OO99T ( = LMG 26739T = HAMBI 3284T) as the type strain. The DNA G+C content is 62.6 mol%. PMID:24969302

  18. Cloning and characterisation of Schistosoma japonicum insulin receptors.

    Directory of Open Access Journals (Sweden)

    Hong You

    2010-03-01

    Full Text Available Schistosomes depend for growth and development on host hormonal signals, which may include the insulin signalling pathway. We cloned and assessed the function of two insulin receptors from Schistosoma japonicum in order to shed light on their role in schistosome biology.We isolated, from S. japonicum, insulin receptors 1 (SjIR-1 and 2 (SjIR-2 sharing close sequence identity to their S. mansoni homologues (SmIR-1 and SmIR-2. SjIR-1 is located on the tegument basal membrane and the internal epithelium of adult worms, whereas SjIR-2 is located in the parenchyma of males and the vitelline tissue of females. Phylogenetic analysis showed that SjIR-2 and SmIR-2 are close to Echinococcus multilocularis insulin receptor (EmIR, suggesting that SjIR-2, SmIR-2 and EmIR share similar roles in growth and development in the three taxa. Structure homology modelling recovered the conserved structure between the SjIRs and Homo sapiens IR (HIR implying a common predicted binding mechanism in the ligand domain and the same downstream signal transduction processing in the tyrosine kinase domain as in HIR. Two-hybrid analysis was used to confirm that the ligand domains of SjIR-1 and SjIR-2 contain the insulin binding site. Incubation of adult worms in vitro, both with a specific insulin receptor inhibitor and anti-SjIRs antibodies, resulted in a significant decrease in worm glucose levels, suggesting again the same function for SjIRs in regulating glucose uptake as described for mammalian cells.Adult worms of S. japonicum possess insulin receptors that can specifically bind to insulin, indicating that the parasite can utilize host insulin for development and growth by sharing the same pathway as mammalian cells in regulating glucose uptake. A complete understanding of the role of SjIRs in the biology of S. japonicum may result in their use as new targets for drug and vaccine development against schistosomiasis.

  19. Deletion of the SACPD-C locus alters the symbiotic relationship between Bradyrhizobium japonicum USDA110 and soybean, resulting in elicitation of plant defense response and nodulation defects

    Science.gov (United States)

    Legumes form symbiotic association with soil-dwelling bacteria collectively called rhizobia. This association results in the formation of nodules, unique plant-derived organs, within which the rhizobia are housed. Rhizobia encoded-nitrogenase facilitates the conversation of atmospheric nitrogen int...

  20. Expressão dos genes nodC, nodW e nopP em Bradyrhizobium japonicum estirpe CPAC 15 avaliada por RT-qPCR

    OpenAIRE

    BORTOLAN, S.; Barcellos, F. G; MARCELINO, F. C.; HUNGRIA, M.

    2009-01-01

    O objetivo deste trabalho foi avaliar a expressão, por RT-qPCR, dos genes de nodulação nodC e nodW e do gene nopP da estirpe CPAC 15, que provavelmente atuam na infecção das raízes da soja. Foram realizados dois experimentos. No primeiro, a expressão dos genes foi avaliada nas células após a incubação com genisteína por 15 min, 1, 4 e 8 horas. Os resultados revelaram que os três genes apresentaram maior expressão imediatamente após o contato com o indutor (15 min). No segundo experimento, a b...

  1. Deletion of the SACPD-C Locus Alters the Symbiotic Relationship Between Bradyrhizobium japonicum USDA110 and Soybean, Resulting in Elicitation of Plant Defense Response and Nodulation Defects.

    Science.gov (United States)

    Krishnan, Hari B; Alaswad, Alaa A; Oehrle, Nathan W; Gillman, Jason D

    2016-11-01

    Legumes form symbiotic associations with soil-dwelling bacteria collectively called rhizobia. This association results in the formation of nodules, unique plant-derived organs, within which the rhizobia are housed. Rhizobia-encoded nitrogenase facilitates the conversion of atmospheric nitrogen into ammonia, which is utilized by the plants for its growth and development. Fatty acids have been shown to play an important role in root nodule symbiosis. In this study, we have investigated the role of stearoyl-acyl carrier protein desaturase isoform C (SACPD-C), a soybean enzyme that catalyzes the conversion of stearic acid into oleic acid, which is expressed in developing seeds and in nitrogen-fixing nodules. In-depth cytological investigation of nodule development in sacpd-c mutant lines M25 and MM106 revealed gross anatomical alteration in the sacpd-c mutants. Transmission electron microscopy observations revealed ultrastructural alterations in the sacpd-c mutants that are typically associated with plant defense response to pathogens. In nodules of two sacpd-c mutants, the combined jasmonic acid (JA) species (JA and the isoleucine conjugate of JA) were found to be reduced and 12-oxophytodienoic acid (OPDA) levels were significantly higher relative to wild-type lines. Salicylic acid levels were not significantly different between genotypes, which is divergent from previous studies of sacpd mutant studies on vegetative tissues. Soybean nodule phytohormone profiles were very divergent from those of roots, and root profiles were found to be almost identical between mutant and wild-type genotypes. The activities of antioxidant enzymes, ascorbate peroxidase, and superoxide dismutase were also found to be higher in nodules of sacpd-c mutants. PR-1 gene expression was extremely elevated in M25 and MM106, while the expression of nitrogenase was significantly reduced in these sacpd-c mutants, compared with the parent 'Bay'. Two-dimensional gel electrophoresis and matrix-assisted laser desorption-ionization time of flight mass spectrometry analyses confirmed sacpd-c mutants also accumulated higher amounts of pathogenesis-related proteins in the nodules. Our study establishes a major role for SACPD-C activity as essential for proper maintenance of soybean nodule morphology and physiology and indicates that OPDA signaling is likely to be involved in attenuation of nodule biotic defense responses.

  2. Labeling of carbon pools in Bradyrhizobium japonicum bacteroids in intact nodules following feedings of sup 14 CO sub 2 for periods of <= six minutes

    Energy Technology Data Exchange (ETDEWEB)

    Salminen, S.; Streeter, J. (Ohio State Univ., Wooster (United States))

    1991-05-01

    A system was established for isolation of bacteroids from soybean nodules in 5 min by centrifugation of crude brei through silicone oil in a microfuge. Using marker enzymes {Beta}-hydroxybutyrate dehydrogenase (DH) and cytochrome C oxidase, it was found that bacteroids were not ruptured during isolation, that mitochondrial contamination was {<=} 2%, and that bacteroid recovery was {approx} 93%. About 40 {mu}Ci {sup 14}CO{sub 2} were supplied to 300 mg fresh wt of intact nodules to label the 4-carbon acid pools in the host cytoplasm of infected cells. Following incubations of 1 to 6 min label in the cytosol was largely in malate (MAL), as expected. Label in aspartate and citrate was about 20% that in MAL. {sup 14}C in succinate (SUC) was only {approx} 3% of that in MAL, and fumarate was essentially unlabeled indicating limited equilibration of label via fumarase and SUC DH. In bacteroids MAL was most rapidly labeled, but glutamate {sup 14}C was about 50% that in MAL; this is consistent with other results. Aspartate and alanine were labeled at a rate approx 10% that of MAL, and SUC, citrate and fumarate were slowly labeled. Results are consistent with 4-carbon acids (mainly malate) as the main source of reduced carbon for bacteroids.

  3. Comparative Study of Transcriptome Profiles of Mouse Livers and Skins Infected by Fork-Tailed or Non-Fork-Tailed Schistosoma japonicum.

    Science.gov (United States)

    Yang, Yan; He, Jun-Jun; Hu, Shuang; Chang, Hua; Xiang, Xun; Yang, Jian-Fa; Zou, Feng-Cai

    2017-01-01

    Schistosoma japonicum (S. japonicum) is a worldwide spread pathogen which penetrates host skin and then induces several diseases in infected host, such as fibrosis, formation of granulomas, hepatocirrhosis, and hepatomegaly. In present study, for the first time, transcriptomic profiles of mouse livers and skins infected by fork-tailed S. japonicum cercaria or non-fork-tailed S. japonicum cercaria were analyzed by using RNA-seq. The present findings demonstrated that transcriptomic landscapes of livers and skins infected by fork-tailed S. japonicum cercaria or non-fork-tailed S. japonicum cercaria were different. S. japonicum has great influence on hepatic metabolic processes. Fork-tailed S. japonicum cercaria upregulated hepatic metabolic processes, while non-fork-tailed S. japonicum cercaria downregulated hepatic metabolic processes. For the metabolism process or the metabolism enzyme expressional change, the pharmacokinetics of host could be changed during S. japonicum infection, regardless the biotypes of S. japonicum cercariae. The changes of infected skins focused on upregulation of immune response. During the S. japonicum skin infection period, fork-tailed S. japonicum cercaria infection induced stronger immune response comparing with that immune response triggered by non-fork-tailed S. japonicum cercaria. The transcription factor enrichment analysis showed that Irf7, Stat1 and Stat2 could play important roles in gene regulation during fork-tailed S. japonicum cercaria infection.

  4. Comparative Study of Transcriptome Profiles of Mouse Livers and Skins Infected by Fork-Tailed or Non-Fork-Tailed Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    Yan Yang

    2017-08-01

    Full Text Available Schistosoma japonicum (S. japonicum is a worldwide spread pathogen which penetrates host skin and then induces several diseases in infected host, such as fibrosis, formation of granulomas, hepatocirrhosis, and hepatomegaly. In present study, for the first time, transcriptomic profiles of mouse livers and skins infected by fork-tailed S. japonicum cercaria or non-fork-tailed S. japonicum cercaria were analyzed by using RNA-seq. The present findings demonstrated that transcriptomic landscapes of livers and skins infected by fork-tailed S. japonicum cercaria or non-fork-tailed S. japonicum cercaria were different. S. japonicum has great influence on hepatic metabolic processes. Fork-tailed S. japonicum cercaria upregulated hepatic metabolic processes, while non-fork-tailed S. japonicum cercaria downregulated hepatic metabolic processes. For the metabolism process or the metabolism enzyme expressional change, the pharmacokinetics of host could be changed during S. japonicum infection, regardless the biotypes of S. japonicum cercariae. The changes of infected skins focused on upregulation of immune response. During the S. japonicum skin infection period, fork-tailed S. japonicum cercaria infection induced stronger immune response comparing with that immune response triggered by non-fork-tailed S. japonicum cercaria. The transcription factor enrichment analysis showed that Irf7, Stat1 and Stat2 could play important roles in gene regulation during fork-tailed S. japonicum cercaria infection.

  5. Epidemic Spread of Symbiotic and Non-Symbiotic Bradyrhizobium Genotypes Across California.

    Science.gov (United States)

    Hollowell, A C; Regus, J U; Gano, K A; Bantay, R; Centeno, D; Pham, J; Lyu, J Y; Moore, D; Bernardo, A; Lopez, G; Patil, A; Patel, S; Lii, Y; Sachs, J L

    2016-04-01

    The patterns and drivers of bacterial strain dominance remain poorly understood in natural populations. Here, we cultured 1292 Bradyrhizobium isolates from symbiotic root nodules and the soil root interface of the host plant Acmispon strigosus across a >840-km transect in California. To investigate epidemiology and the potential role of accessory loci as epidemic drivers, isolates were genotyped at two chromosomal loci and were assayed for presence or absence of accessory "symbiosis island" loci that encode capacity to form nodules on hosts. We found that Bradyrhizobium populations were very diverse but dominated by few haplotypes-with a single "epidemic" haplotype constituting nearly 30 % of collected isolates and spreading nearly statewide. In many Bradyrhizobium lineages, we inferred presence and absence of the symbiosis island suggesting recurrent evolutionary gain and or loss of symbiotic capacity. We did not find statistical phylogenetic evidence that the symbiosis island acquisition promotes strain dominance and both symbiotic and non-symbiotic strains exhibited population dominance and spatial spread. Our dataset reveals that a strikingly few Bradyrhizobium genotypes can rapidly spread to dominate a landscape and suggests that these epidemics are not driven by the acquisition of accessory loci as occurs in key human pathogens.

  6. Whole-Genome Sequence of Bradyrhizobium elkanii Strain UASWS1016, a Potential Symbiotic Biofertilizer for Agriculture.

    Science.gov (United States)

    Crovadore, Julien; Calmin, Gautier; Chablais, Romain; Cochard, Bastien; Schulz, Torsten; Lefort, François

    2016-10-06

    Bradyrhizobium elkanii UASWS1016 has been isolated from a wet oxidation sewage plant in Italy. Fully equipped for ammonia assimilation, heavy metal resistances, and aromatic compounds degradation, it carries a large type IV secretion system, specific of plant-associated microbes. Deprived of toxins, it could be considered for agricultural and environmental uses. Copyright © 2016 Crovadore et al.

  7. Synthesis of bradyrhizose oligosaccharides relevant to the Bradyrhizobium O-antigen

    DEFF Research Database (Denmark)

    Li, Wei; Silipo, Alba; Gersby, Lotte Bettina Andersen

    2017-01-01

    The unique α-(1→7)-bradyrhizoside linkages are constructed for the first time via judicious choice of the glycosylation partners and conditions, thus tetra- and penta-bradyrhizosides relevant to the peculiar O-antigen of Bradyrhizobium are synthesized, which are shown to adopt the defined right...

  8. Congenital infection with Schistosoma japonicum but not with Schistosoma bovis in sheep.

    Science.gov (United States)

    Johansen, M V; lburg, T; Morad, J; Ornbjerg, N

    2002-04-01

    The present study investigated whether Schistosoma japonicum or Schistosoma bois could establish prenatally in lambs. Three ewes were exposed to S. japonicum by intramuscular injection of cercariae, and 3 ewes were exposed to S. bovis cercariae using the leg-emerging technique approximately 2 mo before delivery, and 1 age-matched pregnant ewe served as an uninfected control. The study lasted 18-20 wk after infection, which was 8-9 wk after delivery. All 6 exposed ewes became infected with either S. bovis or S. japonicum. Eight lambs were borne by the 7 ewes, of which 1 (S. bovis exposed) was dead and 1 (S. japonicum exposed) died at delivery. Of the 3 S. japonicum-exposed lambs, 2 were found infected. Four lambs born of S. bovis-exposed ewes were negative. Despite having no worms, these 4 S. bovis-exposed lambs as well as the 1 negative S. japonicum-exposed lamb had, in contrast to the nonexposed control lamb, few, but distinct, liver granulomas dominated by eosinophils and giant cells with large central necrotic areas but with no remnants of eggs or worms. Hence, congenital infection was demonstrated in S. japonicum-infected lambs, but not in S. bovis-infected ones.

  9. Follicular helper T cells promote liver pathology in mice during Schistosoma japonicum infection.

    Science.gov (United States)

    Chen, Xiaojun; Yang, Xiaowei; Li, Yong; Zhu, Jifeng; Zhou, Sha; Xu, Zhipeng; He, Lei; Xue, Xue; Zhang, Weiwei; Dong, Xiaoxiao; Wu, Henry; Li, Carrie J; Hsu, Hsiang-Ting; Kong, Wenjun; Liu, Feng; Tripathi, Prem B; Yu, Michelle S; Chang, Jason; Zhou, Liang; Su, Chuan

    2014-05-01

    Following Schistosoma japonicum (S. japonicum) infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease. While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host. However, the exact mechanism of hepatic granuloma formation has remained obscure. In this study, we for the first time showed that follicular helper T (Tfh) cells are recruited to the liver to upregulate hepatic granuloma formation and liver injury in S. japonicum-infected mice, and identified a novel function of macrophages in Tfh cell induction. In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL) on macrophages which is regulated by CD40-CD40L signaling. Our findings uncovered a previously unappreciated role for Tfh cells in liver pathology caused by S. japonicum infection in mice.

  10. A REVIEW OF SCHISTOSOMIASIS IN INDONESIA WITH REFERENCE TO SCHISTOSOMA JAPONICUM

    Directory of Open Access Journals (Sweden)

    Lim Boo Liat

    2012-09-01

    Full Text Available Tinjauan tentang Schistosoma di Indonesia ini mencakup Schistosoma japonicum, S. incognitum, S. spindale dan Trichohilharzia brevis. Tinjauan dibuat atas dasar laporan-laporan penelitian yang telah diterbitkan. Di dalamnya dapat dijumpai uraian singkat tentang S. spinale dan T. brevis. Dari banyak publikasi tentang S. japonicum dan S. incoganitum dapat disajikan uraian tentang peranan kedua parasit tersebut sebagai penyebab penyakit baik manusia maupun hewan.

  11. Biology and Control of Snail Intermediate Host of Schistosoma japonicum in The People's Republic of China

    DEFF Research Database (Denmark)

    Li, Z.J.; Ge, J; Dai, J.R.

    2016-01-01

    Schistosomiasis caused by Schistosoma japonicum is a severe parasitic disease in The People's Republic of China and imposed considerable burden on human and domestic animal health and socioeconomic development. The significant achievement in schistosomiasis control has been made in last 60years. ....... Oncomelania hupensis as the only intermediate host of S. japonicum plays a key role in disease transmission. The habitat complexity of the snails challenges to effective control. In this review we share the experiences in control and research of O. hupensis....

  12. A specific indel marker for the Philippines Schistosoma japonicum revealed by analysis of mitochondrial genome sequences.

    Science.gov (United States)

    Li, Juan; Chen, Fen; Sugiyama, Hiromu; Blair, David; Lin, Rui-Qing; Zhu, Xing-Quan

    2015-07-01

    In the present study, near-complete mitochondrial (mt) genome sequences for Schistosoma japonicum from different regions in the Philippines and Japan were amplified and sequenced. Comparisons among S. japonicum from the Philippines, Japan, and China revealed a geographically based length difference in mt genomes, but the mt genomic organization and gene arrangement were the same. Sequence differences among samples from the Philippines and all samples from the three endemic areas were 0.57-2.12 and 0.76-3.85 %, respectively. The most variable part of the mt genome was the non-coding region. In the coding portion of the genome, protein-coding genes varied more than rRNA genes and tRNAs. The near-complete mt genome sequences for Philippine specimens were identical in length (14,091 bp) which was 4 bp longer than those of S. japonicum samples from Japan and China. This indel provides a unique genetic marker for S. japonicum samples from the Philippines. Phylogenetic analyses based on the concatenated amino acids of 12 protein-coding genes showed that samples of S. japonicum clustered according to their geographical origins. The identified mitochondrial indel marker will be useful for tracing the source of S. japonicum infection in humans and animals in Southeast Asia.

  13. Schistosoma japonicum: immunological characterization and detection of circulating polysaccharide antigens from adult worms.

    Science.gov (United States)

    Qian, Z L; Deelder, A M

    1983-04-01

    The antigenic constituents of a trichloroacetic acid (TCA)-soluble fraction of adult Schistosoma japonicum were studied with immunoelectrophoresis, and compared with those of Schistosoma mansoni. Eight TCA-soluble antigens of S. japonicum were demonstrated, five of which showed immunological identity with S. mansoni antigens. Of the eight antigens, five antigens with anodic motility were found as circulating antigens in S. japonicum-infected hamster and rabbit sera; the major circulating antigen was the circulating anodic antigen (CAA). Two other antigens, with cathodic motility, including the circulating cathodic antigen (CCA), were demonstrable as circulating antigens in S. mansoni infections, but not in S. japonicum infections. Most of the circulating antigens were shown to be gut-associated. Only one antigen, line 2, which was not demonstrable as circulating antigen and which was present in the parenchyma of the worms, was found to be specific for S. japonicum. Using an ELISA for the detection of CAA in the sera of S. japonicum-infected rabbits, a lower detection level of 100 ng CAA/ml serum was achieved. Moreover, at 7-8 weeks after infection, a direct relationship between worm burden and CAA level was demonstrated.

  14. Origin and Evolution of Nitrogen Fixation Genes on Symbiosis Islands and Plasmid in Bradyrhizobium.

    Science.gov (United States)

    Okubo, Takashi; Piromyou, Pongdet; Tittabutr, Panlada; Teaumroong, Neung; Minamisawa, Kiwamu

    2016-09-29

    The nitrogen fixation (nif) genes of nodule-forming Bradyrhizobium strains are generally located on symbiosis islands or symbiosis plasmids, suggesting that these genes have been transferred laterally. The nif genes of rhizobial and non-rhizobial Bradyrhizobium strains were compared in order to infer the evolutionary histories of nif genes. Based on all codon positions, the phylogenetic tree of concatenated nifD and nifK sequences showed that nifDK on symbiosis islands formed a different clade from nifDK on non-symbiotic loci (located outside of symbiosis islands and plasmids) with elongated branches; however, these genes were located in close proximity, when only the 1st and 2nd codon positions were analyzed. The guanine (G) and cytosine (C) content of the 3rd codon position of nifDK on symbiosis islands was lower than that on non-symbiotic loci. These results suggest that nif genes on symbiosis islands were derived from the non-symbiotic loci of Bradyrhizobium or closely related strains and have evolved toward a lower GC content with a higher substitution rate than the ancestral state. Meanwhile, nifDK on symbiosis plasmids clustered with nifDK on non-symbiotic loci in the tree representing all codon positions, and the GC content of symbiotic and non-symbiotic loci were similar. These results suggest that nif genes on symbiosis plasmids were derived from the non-symbiotic loci of Bradyrhizobium and have evolved with a similar evolutionary pattern and rate as the ancestral state.

  15. Thioredoxin glutathione reductase as a novel drug target: evidence from Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    LiJun Song

    Full Text Available BACKGROUND: Schistosomiasis remains a major public health concern affecting billions of people around the world. Currently, praziquantel is the only drug of choice for treatment of human schistosomiasis. The emergence of drug resistance to praziquantel in schistosomes makes the development of novel drugs an urgent task. Thioredoxin glutathione reductase (TGR enzymes in Schistosoma mansoni and some other platyhelminths have been identified as alternative targets. The present study was designed to confirm the existense and the potential value of TGR as a target for development of novel antischistosomal agents in Schistosoma japonicum, a platyhelminth endemic in Asia. METHODS AND FINDINGS: After cloning the S. japonicum TGR (SjTGR gene, the recombinant SjTGR selenoprotein was purified and characterized in enzymatic assays as a multifunctional enzyme with thioredoxin reductase (TrxR, glutathione reductase (GR and glutaredoxin (Grx activities. Immunological and bioinformatic analyses confirmed that instead of having separate TrxR and GR proteins in mammalian, S. japonicum only encodes TGR, which performs the functions of both enzymes and plays a critical role in maintaining the redox balance in this parasite. These results were in good agreement with previous findings in Schistosoma mansoni and some other platyhelminths. Auranofin, a known inhibitor against TGR, caused fatal toxicity in S. japonicum adult worms in vitro and reduced worm and egg burdens in S. japonicum infected mice. CONCLUSIONS: Collectively, our study confirms that a multifunctional enzyme SjTGR selenoprotein, instead of separate TrxR and GR enzymes, exists in S. japonicum. Furthermore, TGR may be a potential target for development of novel agents against schistosomes. This assumption is strengthened by our demonstration that the SjTGR is an essential enzyme for maintaining the thiol-disulfide redox homeostasis of S. japonicum.

  16. Transmission dynamics of Schistosoma japonicum in the lakes and marshlands of China.

    Directory of Open Access Journals (Sweden)

    Darren J Gray

    Full Text Available Schistosoma japonicum is a major public health concern in China, with over one million people infected and another 40 million living in areas at risk of infection. Unlike the disease caused by S. mansoni and S. haematobium, schistosomiasis japonica is a zoonosis, involving a number of different mammalian species as reservoir hosts. As a result of a number of published reports from China, it has long been considered that bovines, particularly water buffaloes, play a major role in human S. japonicum transmission there, and a drug-based intervention study (1998-2003 around the Poyang Lake in Jiangxi Province provided proof of concept that water buffaloes are, indeed, major reservoirs of human infection in this setting.In this study we incorporated recently obtained epidemiological information to model the steady-state S. japonicum transmission as well as the impact of the removal of S. japonicum transmission attributable to water buffaloes on human infection rates across six different endemic scenarios within three villages in the Dongting (Hunan and Poyang (Jiangxi lakes of southern China. Similar results were obtained for all scenarios. Steady-state S. japonicum infection rates remained constant and human prevalence and incidence were predicted to fall considerably over time. The model showed that the contribution of S. japonicum water buffalo transmission to human infection ranged from 39.1% to 99.1% and predicted that the removal of water buffalo transmission would reduce parasite reproductive rates below 1. This indicates that without the contribution of water buffaloes, S. japonicum transmission is interrupted and unsustainable. These scenarios are generalizable to other endemic villages in the lake and marshland areas of China where a similar cycle of snail infection and infection/reinfection of humans and bovines occurs.Along with previous epidemiological data, our findings strongly support water buffaloes as an important component of the

  17. Ganoderma neo-japonicum Imazeki revisited: Domestication study and antioxidant properties of its basidiocarps and mycelia.

    Science.gov (United States)

    Tan, Wee-Cheat; Kuppusamy, Umah Rani; Phan, Chia-Wei; Tan, Yee-Shin; Raman, Jegadeesh; Anuar, Azliza Mad; Sabaratnam, Vikineswary

    2015-07-27

    Mushroom cultivation benefits humankind as it deliberately encourages wild mushrooms to be commercially propagated while recycling agricultural wastes. Ganoderma neo-japonicum is a rare polypore mushroom found growing on decaying Schizostachyum brachycladium (a tropical bamboo) clumps in Malaysia. The Malaysian indigenous tribes including the Temuans and Temiars use the basidiocarps of G. neo-japonicum to treat various ailments including diabetes. In this study, the domestication of G. neo-japonicum in artificial logs of different agricultural residues was investigated. Sawdust promoted the mycelia spawn colonisation in the shortest period of 38 ± 0.5 days. However, only sawdust and bamboo dust supported the primodia formation. Complex medium supported mycelium growth in submerged cultures and 27.11 ± 0.43 g/L of mycelia was obtained after 2 weeks of cultivation at 28 °C and 200 rpm. Antioxidant potential in mushroom may be influenced by different cultivation and extraction methods. The different extracts from the wild and cultivated basidiocarps as well as mycelia were then tested for their antioxidant properties. Aqueous and ethanol extracts of mycelia and basidiocarps tested had varying levels of antioxidant activities. To conclude, domestication of wild G. neo-japonicum using agroresidues may ensure a continuous supply of G. neo-japonicum for its medicinal use while ensuring the conservation of this rare species.

  18. Genetic diversity and selection of three nuclear genes in Schistosoma japonicum populations.

    Science.gov (United States)

    Li, Yaqi; Yin, Mingbo; Wu, Qunfeng; McManus, Donald P; Blair, David; Li, Hongyan; Xu, Bin; Mo, Xiaojin; Feng, Zheng; Hu, Wei

    2017-02-17

    The blood fluke, Schistosoma japonicum still causes severe disease in China, the Philippines and Indonesia. Although there have been some studies the molecular epidemiology of this persistent and harmful parasite, few have explored the possibility and implications of selection in S. japonicum populations. We analyzed diversity and looked for evidence of selection at three nuclear genes (SjIpp2, SjFabp and SjT22.6) in 13 S. japonicum populations. SjT22.6 was found to exhibit high nucleotide diversity and was under positive selection in the mountainous region of mainland China. As a tegumental protein, its secondary and tertiary structure differed between S. japonicum strains from the mountainous and lakes regions. In contrast, SjIpp2 and SjFabp had relatively low levels of nucleotide diversity and did not show significant departure from neutrality. As a tegument-associated antigen-encoding gene of S. japonicum, SjT22.6 has high nucleotide diversity and appears to be under positive selection in the mountainous region of mainland China.

  19. Novel real-time PCr for detection of Schistosoma japonicum in stool.

    Science.gov (United States)

    Lier, T; Simonsen, G S; Haaheim, H; Hjelmevoll, S O; Vennervald, B J; Johansen, M V

    2006-03-01

    Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples.

  20. A piezoelectric immunosensor using hybrid self-assembled monolayers for detection of Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Shiping Wang

    Full Text Available BACKGROUND: The parasite Schistosoma japonicum causes schistosomiasis disease, which threatens human life and hampers economic and social development in some Asian countries. An important lesson learned from efforts to reduce the occurrence of schistosomiasis is that the diagnostic approach must be altered as further progress is made towards the control and ultimate elimination of the disease. METHODOLOGY/PRINCIPAL FINDINGS: Using mixed self-assembled monolayer membrane (mixed SAM technology, a mixture of mercaptopropionic acid (MPA and mercaptoethanol (ME was self-assembled on the surface of quartz crystals by gold-sulphur-bonds. Soluble egg antigens (SEA of S. japonicum were then cross-linked to the quartz crystal using a special coupling agent. As compared with the traditional single self-assembled monolayer immobilization method, S. japonicum antigen (SjAg immobilization using mixed self-assembled monolayers exhibits much greater immunoreactivity. Under optimal experimental conditions, the detection range is 1:1500 to 1:60 (infected rabbit serum dilution ratios. We measured several infected rabbit serum samples with varying S. japonicum antibody (SjAb concentrations using both immunosensor and ELISA techniques and then produced a correlation analysis. The correlation coefficients reached 0.973. CONCLUSIONS/SIGNIFICANCE: We have developed a new, simple, sensitive, and reusable piezoelectric immunosensor that directly detects SjAb in the serum. This method may represent an alternative to the current diagnostic methods for S. japonicum infection in the clinical laboratory or for analysis outside the laboratory.

  1. An insight into the genetic variation of Schistosoma japonicum in mainland China using DNA microsatellite markers.

    Science.gov (United States)

    Shrivastava, Jaya; Qian, Bao Zhen; Mcvean, Gilean; Webster, Joanne P

    2005-03-01

    This study presents the first microsatellite investigation into the level of genetic variation among Schistosoma japonicum from different geographical origins. S. japonicum isolates were obtained from seven endemic provinces across mainland China: Zhejiang (Jiashan County), Anhui (Guichi County), Jiangxi (Yongxiu County), Hubei (Wuhan County), Hunan (Yueyang area), Sichuan 1 (Maoshan County), Sichuan 2 (Tianquan County), Yunnan (Dali County), and also one province in the Philippines (Sorsogon). DNA from 20 individuals from each origin were screened against 11 recently isolated and characterized S. japonicum microsatellites, and a set of nine loci were selected based on their polymorphic information content. High levels of polymorphism were obtained between and within population samples, with Chinese and Philippine strains appearing to follow different lineages, and with distinct branching between provinces. Moreover, across mainland China, genotype clustering appeared to be related to habitat type and/or intermediate host morph. These results highlight the suitability of microsatellites for population genetic studies of S. japonicum and suggest that there may be different strains of S. japonicum circulating in mainland China.

  2. Ganoderma neo-japonicum Imazeki revisited: Domestication study and antioxidant properties of its basidiocarps and mycelia

    Science.gov (United States)

    Tan, Wee-Cheat; Kuppusamy, Umah Rani; Phan, Chia-Wei; Tan, Yee-Shin; Raman, Jegadeesh; Anuar, Azliza Mad; Sabaratnam, Vikineswary

    2015-01-01

    Mushroom cultivation benefits humankind as it deliberately encourages wild mushrooms to be commercially propagated while recycling agricultural wastes. Ganoderma neo-japonicum is a rare polypore mushroom found growing on decaying Schizostachyum brachycladium (a tropical bamboo) clumps in Malaysia. The Malaysian indigenous tribes including the Temuans and Temiars use the basidiocarps of G. neo-japonicum to treat various ailments including diabetes. In this study, the domestication of G. neo-japonicum in artificial logs of different agricultural residues was investigated. Sawdust promoted the mycelia spawn colonisation in the shortest period of 38 ± 0.5 days. However, only sawdust and bamboo dust supported the primodia formation. Complex medium supported mycelium growth in submerged cultures and 27.11 ± 0.43 g/L of mycelia was obtained after 2 weeks of cultivation at 28 °C and 200 rpm. Antioxidant potential in mushroom may be influenced by different cultivation and extraction methods. The different extracts from the wild and cultivated basidiocarps as well as mycelia were then tested for their antioxidant properties. Aqueous and ethanol extracts of mycelia and basidiocarps tested had varying levels of antioxidant activities. To conclude, domestication of wild G. neo-japonicum using agroresidues may ensure a continuous supply of G. neo-japonicum for its medicinal use while ensuring the conservation of this rare species. PMID:26213331

  3. Cross-sectional associations between intensity of animal and human infection with Schistosoma japonicum in Western Samar province, Philippines

    DEFF Research Database (Denmark)

    McGarvey, Stephen T.; Carabin, Hélène; Batalong, Ernesto Jr.

    2006-01-01

    To estimate the association between the intensity of animal infection with Schistosoma japonicum and human infection in Western Samar province, the Philippines......To estimate the association between the intensity of animal infection with Schistosoma japonicum and human infection in Western Samar province, the Philippines...

  4. In vivo and in vitro activity of oil extract of garlic (Allium sativum Linnaeus) against Schistosoma japonicum cercariae.

    Science.gov (United States)

    Wan, Kang; Wang, Peng; Zhang, Leibo

    2017-01-01

    The activity of garlic oil extract against Schistosoma japonicum cercariae was evaluated. The in vitro and in vivo cercaricidal activities against S. japonicum larvae were determined. Exposure to ≥ 10-6 (v/v) garlic emulsions for 30 min led to 100% cercariae mortality; pre-exposure treatment with ≥ 10-4 (v/v) garlic emulsions showed 100% preventive efficacy against S. japonicum infection, while pre-treatment with 10-5 and 10-6 (v/v) emulsions achieved 20%-40% preventive efficacy and 35.2%-63.6% worm burden reduction. Garlic oil extract has activity against S. japonicum larvae and a promising preventive efficacy against S. japonicum infection.

  5. High prevalence of Schistosoma japonicum infection in Carabao from Samar Province, the Philippines: implications for transmission and control.

    Directory of Open Access Journals (Sweden)

    Catherine A Gordon

    Full Text Available Schistosoma japonicum is endemic in the Philippines, China and Indonesia, and infects more than 40 mammalian host species, all of which can act as reservoirs of infection. In China, water buffaloes have been shown to be major reservoirs of human infection. However, in the Philippines, carabao have not been considered important reservoir hosts for S. japonicum due to the low prevalence and infection intensities reported, the only exception being a qPCR-based study indicating 51% of carabao were S. japonicum-positive. However, the low prevalence found for the same animals when using conventional copro-parasitological techniques means that there is still confusion about the role of carabao in the transmission of schistosomiasis japonicum. To address this inconsistency, and to shed light on the potential role of carabao in the transmission of S. japonicum in the Philippines, we undertook a pilot survey, collecting fecal samples from animals in Western Samar Province and we used a combination of molecular and copro-parasitological techniques to determine the prevalence and intensity of S. japonicum. We found a high prevalence of S. japonicum in the carabao using a validated real-time PCR (qPCR and a copro-parasitological tool, the formalin-ethyl acetate sedimentation (FEA-SD technique. A much lower prevalence of S. japonicum was recorded for the same fecal samples using conventional PCR, the Kato-Katz technique and miracidial hatching. These results suggest that, due to their low diagnostic sensitivity, traditional copro-parasitological techniques underestimate infection in carabao. The use of FEA-SD and qPCR provides a more accurate diagnosis. Based on these findings, the role of bovines in the transmission of S. japonicum appears to be more important in the Philippines than previously recognized, and this may have significant implications for the future control of schistosomiasis there, particularly as, in contrast with previous surveys, we found

  6. Evolutionary and biomedical implications of a Schistosoma japonicum complementary DNA resource.

    Science.gov (United States)

    Hu, Wei; Yan, Qing; Shen, Da-Kang; Liu, Feng; Zhu, Zhi-Dong; Song, Huai-Dong; Xu, Xiang-Ru; Wang, Zhao-Jun; Rong, Yi-Ping; Zeng, Ling-Chun; Wu, Jian; Zhang, Xin; Wang, Ju-Jun; Xu, Xue-Nian; Wang, Sheng-Yue; Fu, Gang; Zhang, Xiang-Lin; Wang, Zhi-Qin; Brindley, Paul J; McManus, Donald P; Xue, Chun-Liang; Feng, Zheng; Chen, Zhu; Han, Ze-Guang

    2003-10-01

    Schistosoma japonicum causes schistosomiasis in humans and livestock in the Asia-Pacific region. Knowledge of the genome of this parasite should improve understanding of schistosome-host interactions, biomedical aspects of schistosomiasis and invertebrate evolution. We assigned 43,707 expressed sequence tags (ESTs) derived from adult S. japonicum and their eggs to 13,131 gene clusters. Of these, 35% shared no similarity with known genes and 75% had not been reported previously in schistosomes. Notably, S. japonicum encoded mammalian-like receptors for insulin, progesterone, cytokines and neuropeptides, suggesting that host hormones, or endogenous parasite homologs, could orchestrate schistosome development and maturation and that schistosomes modulate anti-parasite immune responses through inhibitors, molecular mimicry and other evasion strategies.

  7. Development of chiral praziquantel analogues as potential drug candidates with activity to juvenile Schistosoma japonicum.

    Science.gov (United States)

    Zheng, Yang; Dong, LanLan; Hu, Changyan; Zhao, Bo; Yang, Chunhua; Xia, Chaoming; Sun, Dequn

    2014-09-01

    A series of chiral praziquantel analogues were synthesized and evaluated against Schistosoma japonicum both in vitro and in vivo. All compounds exhibited low to considerable good activity in vivo. Remarkably, worm reduction rate of R-3 was 60.0% at a single oral dose of 200mg/kg against juvenile stage of Schistosoma japonicum. The target compounds displayed in vivo antischistosomal activity against both Schistosoma japonicum and Schistosoma mansoni. Furthermore, all R-isomers displayed stronger antischistosomal activity than S-isomers in vivo, indicating R-isomers were the active enantiomers, while S-isomers were less active ones. This structure activity relationship (SAR) could have important implications in further drug development for schistosomiasis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Soybeans inoculated with root zone soils of Canadian native legumes harbour diverse and novel Bradyrhizobium spp. that possess agricultural potential.

    Science.gov (United States)

    Bromfield, Eden S P; Cloutier, Sylvie; Tambong, James T; Tran Thi, Thu Van

    2017-10-01

    An assessment was made of the evolutionary relationships of soybean nodulating bacteria associated with legumes native to eastern Canada to identify potential new sources of soybean inoculant strains. Short season soybeans were used to selectively trap bacteria from root zone soils of four native legume species. Screening of more than 800 bacterial isolates from soybean root nodules by analysis of recA gene sequences followed by analyses of selected genotypes using six core and two symbiosis (nodC and nifH) gene sequences permitted identification of diverse taxa that included eight novel and four named Bradyrhizobium species as well as lineages attributed to the genera Afipia and Tardiphaga. Plant tests showed that symbionts related to four named species as well as a novel Bradyrhizobium lineage were highly efficient with regard to nitrogen fixation on soybeans relative to an inoculant strain. A new symbiovar (sv. septentrionalis) is proposed based on a group of four novel Bradyrhizobium spp. that possess distinctive nodC and nifH gene sequences and symbiotic characteristics. Evidence is provided for horizontal transfer of sv. septentrionalis symbiosis genes between novel Bradyrhizobium spp., a process that rendered recipient bacteria ineffective on soybeans. Diverse lineages of non-symbiotic and symbiotic Bradyrhizobium spp. co-occured within monophyletic clusters in a phylogenetic tree of concatenated core genes, suggesting that loss and/or gain of symbiosis genes has occurred in the evolutionary history of the bacterial genus. Our data suggest that symbiont populations associated with legumes native to eastern Canada harbour elite strains of Bradyrhizobium for soybean inoculation. Crown Copyright © 2017. Published by Elsevier GmbH. All rights reserved.

  9. Members of Microvirga and Bradyrhizobium genera are native endosymbiotic bacteria nodulating Lupinus luteus in Northern Tunisian soils.

    Science.gov (United States)

    Msaddak, Abdelhakim; Rejili, Mokhtar; Durán, David; Rey, Luis; Imperial, Juan; Palacios, Jose Manuel; Ruiz-Argüeso, Tomas; Mars, Mohamed

    2017-06-01

    The genetic diversity of bacterial populations nodulating Lupinus luteus (yellow lupine) in Northern Tunisia was examined. Phylogenetic analyses of 43 isolates based on recA and gyrB partial sequences grouped them in three clusters, two of which belong to genus Bradyrhizobium (41 isolates) and one, remarkably, to Microvirga (2 isolates), a genus never previously described as microsymbiont of this lupine species. Representatives of the three clusters were analysed in-depth by multilocus sequence analysis of five housekeeping genes (rrs, recA, glnII, gyrB and dnaK). Surprisingly, the Bradyrhizobium cluster with the two isolates LluI4 and LluTb2 may constitute a new species defined by a separate position between Bradyrhizobium manausense and B. denitrificans. A nodC-based phylogeny identified only two groups: one formed by Bradyrhizobium strains included in the symbiovar genistearum and the other by the Microvirga strains. Symbiotic behaviour of representative isolates was tested, and among the seven legumes inoculated only a difference was observed i.e. the Bradyrhizobium strains nodulated Ornithopus compressus unlike the two strains of Microvirga. On the basis of these data, we conclude that L. luteus root nodule symbionts in Northern Tunisia are mostly strains within the B. canariense/B. lupini lineages, and the remaining strains belong to two groups not previously identified as L. luteus endosymbionts: one corresponding to a new clade of Bradyrhizobium and the other to the genus Microvirga. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  10. Pyrosequencing for rapid molecular identification of Schistosoma japonicum and S. mekongi eggs and cercariae.

    Science.gov (United States)

    Thanchomnang, Tongjit; Tantrawatpan, Chairat; Intapan, Pewpan M; Sri-Aroon, Pusadee; Limpanont, Yanin; Lulitanond, Viraphong; Janwan, Penchom; Sanpool, Oranuch; Tourtip, Somjintana; Maleewong, Wanchai

    2013-09-01

    Schistosomiasis, which is caused by Schistosoma japonicum and S. mekongi, is a chronic and dangerous widespread disease affecting several countries in Asia. Differentiation between S. japonicum and S. mekongi eggs and/or cercariae via microscopic examination is difficult due to morphological similarities. It is important to identify these etiological agents isolated from animals and humans at the species or genotype level. In this study, a pyrosequencing assay designed to detect S. japonicum and S. mekongi DNA in fecal samples and infected snails was developed and evaluated as an alternative tool to diagnose schistosomiasis. New primers targeting the 18S ribosomal RNA gene were designated for specific amplification. S. japonicum and S. mekongi were identified using a 43-nucleotide pattern of the 18S ribosomal RNA gene and were differentiated using 7 nucleotides within this region. S. japonicum and S. mekongi-infected snails and fecal samples derived from infected mice and rats were differentially detected within a short period of time. The analytical sensitivity of the method enabled the identification of as little as a single cercaria artificially introduced into a pool of 10 non-infected snails and 2 eggs inoculated in 100mg of non-infected fecal sample. To evaluate the comparative efficacy of the assay, identical samples were also analyzed via microscopy and Sanger sequencing. The pyrosequencing technique was found to be superior to the microscopy method and more rapid than the Sanger sequencing method. These results suggest that the pyrosequencing assay is rapid, simple, sensitive and accurate in identifying S. japonicum and S. mekongi in intermediate hosts and fecal samples of the final host. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. SjTPdb: integrated transcriptome and proteome database and analysis platform for Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    Wang Zhi-Qin

    2008-06-01

    Full Text Available Abstract Background Schistosoma japonicum is one of the three major blood fluke species, the etiological agents of schistosomiasis which remains a serious public health problem with an estimated 200 million people infected in 76 countries. In recent years, enormous amounts of both transcriptomic and proteomic data of schistosomes have become available, providing information on gene expression profiles for developmental stages and tissues of S. japonicum. Here, we establish a public searchable database, termed SjTPdb, with integrated transcriptomic and proteomic data of S. japonicum, to enable more efficient access and utility of these data and to facilitate the study of schistosome biology, physiology and evolution. Description All the available ESTs, EST clusters, and the proteomic dataset of S. japonicum are deposited in SjTPdb. The core of the database is the 8,420 S. japonicum proteins translated from the EST clusters, which are well annotated for sequence similarity, structural features, functional ontology, genomic variations and expression patterns across developmental stages and tissues including the tegument and eggshell of this flatworm. The data can be queried by simple text search, BLAST search, search based on developmental stage of the life cycle, and an integrated search for more specific information. A PHP-based web interface allows users to browse and query SjTPdb, and moreover to switch to external databases by the following embedded links. Conclusion SjTPdb is the first schistosome database with detailed annotations for schistosome proteins. It is also the first integrated database of both transcriptome and proteome of S. japonicum, providing a comprehensive data resource and research platform to facilitate functional genomics of schistosome. SjTPdb is available from URL: http://function.chgc.sh.cn/sj-proteome/index.htm.

  12. Complete genome sequence of the mitochondrial DNA of the river lamprey, Lethenteron japonicum.

    Science.gov (United States)

    Kawai, Yuri L; Yura, Kei; Shindo, Miyuki; Kusakabe, Rie; Hayashi, Keiko; Hata, Kenichiro; Nakabayashi, Kazuhiko; Okamura, Kohji

    2015-01-01

    Lampreys are eel-like jawless fishes evolutionarily positioned between invertebrates and vertebrates, and have been used as model organisms to explore vertebrate evolution. In this study we determined the complete genome sequence of the mitochondrial DNA of the Japanese river lamprey, Lethenteron japonicum, using next-generation sequencers. The sequence was 16,272 bp in length. The gene content and order were identical to those of the sea lamprey, Petromyzon marinus, which has been the reference among lamprey species. However, the sequence similarity was less than 90%, suggesting the need for the whole-genome sequencing of L. japonicum.

  13. Sobrevivência de Bradyrhizobium e Azorhizobium em misturas de solo contaminadas com metais pesados

    Directory of Open Access Journals (Sweden)

    A. Matsuda

    2002-03-01

    Full Text Available Estudos foram realizados no Departamento de Ciência do Solo da Universidade Federal de Lavras (MG, no período de novembro/1999 a janeiro/2000, com o objetivo de avaliar a sobrevivência de estirpe e isolados de rizóbio em solo contaminado com metais pesados e verificar a relação entre tolerância do rizóbio a metais pesados em meio de cultura e sua sobrevivência em solo contaminado. Foram utilizados os dois microrganismos mais tolerantes [BR-4406 (estirpe recomendada para Enterolobium spp. e UFLA-01-457 (isolado de solo contaminado, ambos pertencentes ao gênero Bradyrhizobium ] e os dois mais sensíveis (UFLA-01-486 e UFLA-01-510, isolados de solo contaminado, pertencentes ao gênero Azorhizobium , todos selecionados de um grupo de 60estirpes/isolados em estudos prévios deste laboratório, em meio de cultura suplementado com metais pesados.Empregaram-se misturas de um Latossolo Vermelho-Escuro (LE que continham 0, 15 e 45% (v/v de um Latossolo Vermelho-Amarelo plíntico contaminado com Zn, Cd, Pb e Cu. As misturas de solo contaminado foram inoculadas com 20mL de cultura em YM na fase log das estirpes mencionadas, as quais foram testadas separadamente com três repetições. A avaliação do número de células viáveis no solo, realizada aos 0, 7, 14, 21 e 28dias de incubação, pelo método das diluições sucessivas e inoculação em placas com meio YMA, revelou comportamento diferenciado entre os organismos estudados. O número médio de células que sobreviveram ao final de 28 dias de incubação foi de (em UFCg-1de solo: 10(10,36, 10(10,29 e 10(9,70, para Bradyrhizobium, e 10(9,36, 10(7,54 e 0, para Azorhizobium em misturas de 0, 15 e 45% de solo contaminado, respectivamente. Portanto, houve maior sobrevivência de Bradyrhizobium do que de Azorhizobium , indicando maior tolerância a metais pesados do primeiro gênero.Como Bradyrhizobium foi também mais tolerante "in vitro", os resultados indicam haver relação entre o

  14. Nonnodulating Bradyrhizobium spp. Modulate the Benefits of Legume-Rhizobium Mutualism.

    Science.gov (United States)

    Gano-Cohen, Kelsey A; Stokes, Peter J; Blanton, Mia A; Wendlandt, Camille E; Hollowell, Amanda C; Regus, John U; Kim, Deborah; Patel, Seema; Pahua, Victor J; Sachs, Joel L

    2016-09-01

    Rhizobia are best known for nodulating legume roots and fixing atmospheric nitrogen for the host in exchange for photosynthates. However, the majority of the diverse strains of rhizobia do not form nodules on legumes, often because they lack key loci that are needed to induce nodulation. Nonnodulating rhizobia are robust heterotrophs that can persist in bulk soil, thrive in the rhizosphere, or colonize roots as endophytes, but their role in the legume-rhizobium mutualism remains unclear. Here, we investigated the effects of nonnodulating strains on the native Acmispon-Bradyrhizobium mutualism. To examine the effects on both host performance and symbiont fitness, we performed clonal inoculations of diverse nonnodulating Bradyrhizobium strains on Acmispon strigosus hosts and also coinoculated hosts with mixtures of sympatric nodulating and nonnodulating strains. In isolation, nonnodulating Bradyrhizobium strains did not affect plant performance. In most cases, coinoculation of nodulating and nonnodulating strains reduced host performance compared to that of hosts inoculated with only a symbiotic strain. However, coinoculation increased host performance only under one extreme experimental treatment. Nearly all estimates of nodulating strain fitness were reduced in the presence of nonnodulating strains. We discovered that nonnodulating strains were consistently capable of coinfecting legume nodules in the presence of nodulating strains but that the fitness effects of coinfection for hosts and symbionts were negligible. Our data suggest that nonnodulating strains most often attenuate the Acmispon-Bradyrhizobium mutualism and that this occurs via competitive interactions at the root-soil interface as opposed to in planta Rhizobia are soil bacteria best known for their capacity to form root nodules on legume plants and enhance plant growth through nitrogen fixation. Yet, most rhizobia in soil do not have this capacity, and their effects on this symbiosis are poorly

  15. Genome sequence of Bradyrhizobium sp. LMTR 3, a diazotrophic symbiont of Lima bean (Phaseolus lunatus

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    Ernesto Ormeño-Orrillo

    2017-09-01

    Full Text Available Bradyrhizobium sp. LMTR 3 is a representative strain of one of the geno(species of diazotrophic symbionts associated with Lima bean (Phaseolus lunatus in Peru. Its 7.83 Mb genome was sequenced using the Illumina technology and found to encode a complete set of genes required for nodulation and nitrogen fixation, and additional genes putatively involved in root colonization. Its draft genome sequence and annotation have been deposited at GenBank under the accession number MAXC00000000.

  16. Pharmacokinetics and risk evaluation of DNA vaccine against Schistosoma japonicum.

    Science.gov (United States)

    Liu, Hai-Feng; Li, Wei; Lu, Ming-Bo; Yu, Long-Jiang

    2013-01-01

    DNA plasmid immunization is a novel approach of preventive and therapeutic vaccine. More than 100 DNA vaccines have been on preclinical or clinical phase trials, and four kinds of DNA vaccines for livestock have been approved by USDA, CFIA, and APVMA. Schistosomiasis is a worldwide parasitic disease, and vaccine immunization is supposed to be a promising approach to control the health crisis. On the basis of former preclinical studies, we further focused on the pharmacokinetics and risk evaluation of DNA vaccine in vivo. In the present study, enhanced green fluorescent protein (EGFP) report gene was fused with Schistosoma japonicum 23 kDa transmembrane protein antigen gene (Sj23) and constructed into DNA vaccine pVIVO2-Sj23.EGFP. After intramuscularly injecting 100 μg of purified DNA vaccine plasmid to immunizate BALB/c mice, we studied the tissue distribution of DNA plasmid and expressed Sj23.EGFP antigen, the persistence time of elicited antibodies, and the risk of DNA vaccine transferred into intestinal microorganisms. The results showed that DNA vaccine plasmid could be distributed into all tissues of the body after injection; however, only few organs including the injected muscle were detected DNA vaccine at postimmunization until the 100 days by PCR technology; the detection of green fluorescence protein displayed that DNA vaccine could be expressed in almost every tissue and organs; the ELISA assay indicated the immune antibody against Sj23 could persist over 70 days; and the DNA vaccine transferring intestinal flora results was negative. The results indicated that the DNA vaccine has systemic protection and long-lasting effectivity and is safe to intestinal flora.

  17. Induction of species-specific immunity against Schistosoma japonicum by exposure of rats to ultra-violet attenuated cercariae

    Energy Technology Data Exchange (ETDEWEB)

    Moloney, N.A.; Webbe, G.; Hinchcliffe, P.

    1987-02-01

    Single percutaneous immunizations of Fischer rats with 1000 ultra-violet attenuated Schistosoma japonicum cercariae induced 52-88% resistance to challenge 4 weeks later. Increasing this to 3 immunizations induced 90% resistance to challenge, and this level of protection remained undiminished for up to 40 weeks after vaccination. Rats vaccinated with gamma-irradiated S. mansoni cercariae were resistant to challenge with S. mansoni but not S. japonicum. Similarly rats vaccinated with u.v.-attenuated S. japonicum cercariae were not resistant to heterologous challenge. Thus irradiated vaccines are species-specific in both permissive and non-permissive hosts.

  18. Photosynthetic Bradyrhizobium sp. Strain ORS285 Is Capable of Forming Nitrogen-Fixing Root Nodules on Soybeans (Glycine max)

    Science.gov (United States)

    Giraud, Eric; Xu, Lei; Chaintreuil, Clémence; Gargani, Daniel; Gully, Djamel

    2013-01-01

    The ability of photosynthetic Bradyrhizobium strains ORS285 and ORS278 to nodulate soybeans was investigated. While the nod gene-deficient ORS278 strain induced bumps only on soybean roots, the nod gene-containing ORS285 strain formed nitrogen-fixing nodules. However, symbiotic efficiencies differed drastically depending on both the soybean genotype used and the culture conditions tested. PMID:23354704

  19. [Susceptibilities of Oncomelania hupensis snails to Schistosoma japonicum miracidia from different hosts].

    Science.gov (United States)

    Tian, Yue; Wang, Tian-Ping; Wang, Qi-Zhi; Lv, Da-Bing; Yin, Xiao-Mei; Zhou, Li; Wang, Zhen-Li; Wang, Feng-Feng; Wang, Yue; Zhang, Le-Sheng

    2011-08-01

    To understand the susceptibilities of Oncomelania hupensis snails to Schistosoma japonicum miracidia from different hosts. The Schistosoma japonicum eggs from different hosts, such as rabbits, cattle and mice were collected. These eggs were incubated for miracidia, respectively. Each snail from the same site was exposed to 5 miracidia of Schistosoma japonicum from different hosts. The infected snails were fed in the laboratory for two months. Then all the snails were dissected and observed under the dissecting microscope in order to know the infection rate of snails. In the experiment group, the infection rates of snails infected with miracidia from rabbits, cattle and mice were 1.42%, 8.67% and 19.87%, respectively, the mortality rates were 29.5%, 13.5% and 24.5%, respectively. However, the infection rates of snails in the control group were 2.63%, 2.02% and 11.66%, respectively, and the mortality rates were 24.0%, 49.5% and 18.5%, respectively. The susceptibilities of Oncomelania snails to Schistosoma japonicum miracidia from 3 kinds of hosts are significantly different.

  20. Follicular helper T cells promote liver pathology in mice during Schistosoma japonicum infection.

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    Xiaojun Chen

    2014-05-01

    Full Text Available Following Schistosoma japonicum (S. japonicum infection, granulomatous responses are induced by parasite eggs trapped in host organs, particular in the liver, during the acute stage of disease. While excessive liver granulomatous responses can lead to more severe fibrosis and circulatory impairment in chronically infected host. However, the exact mechanism of hepatic granuloma formation has remained obscure. In this study, we for the first time showed that follicular helper T (Tfh cells are recruited to the liver to upregulate hepatic granuloma formation and liver injury in S. japonicum-infected mice, and identified a novel function of macrophages in Tfh cell induction. In addition, our results showed that the generation of Tfh cells driven by macrophages is dependent on cell-cell contact and the level of inducible costimulator ligand (ICOSL on macrophages which is regulated by CD40-CD40L signaling. Our findings uncovered a previously unappreciated role for Tfh cells in liver pathology caused by S. japonicum infection in mice.

  1. Characterization of a novel vaccine candidate and serine proteinase inhibitor from Schistosoma japonicum (Sj serpin).

    Science.gov (United States)

    Yan, Yutao; Liu, Shuxian; Song, Guangcheng; Xu, Yixin; Dissous, Colette

    2005-07-15

    Serine proteinase inhibitors (serpins) represent an important superfamily of endogenous inhibitors that regulate proteolytic events active in a variety of physiological functions. Immunological screening of a Schistosoma japonicum adult worm cDNA expression library with sera of Microtus fortis, a naturally resistant vertebrate host, has identified one clone that encoded for a sequence homologous to those of the serpin superfamily. The full-length sequence encoding S. japonicum serpin (Sj serpin) was amplified from adult worm cDNA by using 5'-RACE-PCR and subsequently cloned into the prokaryotic expression vector pET28c. The full-length Sj serpin fusion-protein with his-tag was expressed in E. coli, purified by affinity chromatography and used to immunize New Zealand white rabbits. Sj serpin is located on the tegument in S. japonicum adult worms. C57BL/6 mice immunized with Sj serpin induced the production of high levels of specific IgE and IgG1 subclass antibodies as well as a marked IL-4 response. Lymphocyte surface marker analysis revealed proliferation of CD19 expressing B cells, indicating a predominant Th2-type response to Sj serpin. Immunized mice developed moderate protection against infection of S. japonicum as demonstrated by a 36 and 39% reduction in the recovery of adult worms and eggs, respectively. These data suggested a role for Sj serpin as a vaccine candidate or as a novel target for anti-schistosome drugs.

  2. Modeling the Dynamics and Control of Transmission of Schistosoma japonicum and S. mekongi in Southeast Asia

    OpenAIRE

    ISHIKAWA, Hirofumi; Ohmae, Hiroshi

    2009-01-01

    A mathematical model for transmission of schistosomes is useful to predict effects of various control measures on suppression of these parasites. This review focuses on epidemiological and environmental factors in Schistosoma japonicum and Schistosoma mekongi infections and recent advances in mathematical models of Schistosoma transmission.

  3. Carriers in electron transport from molecular hydrogen to oxygen in Rhizobium japonicum bacteroids.

    Science.gov (United States)

    Eisbrenner, G; Evans, H J

    1982-03-01

    An investigation has been conducted to identify electron transport carriers that participate in the oxidation of H2 by H2 uptake-positive strains of Rhizobium japonicum bacteroids. We have observed that the reduced form of dibromothymoquinone at a concentration of 0.2 mM strongly inhibited H2 uptake, endogenous respiration, and C2H2 reduction by bacteroid suspensions. Reduced dibromothymoquinone, however, failed to inhibit the transfer of electrons from H2 to methylene blue under anaerobic conditions, indicating that the hydrogenase per se is insensitive to this inhibitor. Metronidazole, at 1 mM, affected rates of H2 uptake and endogenous respiration only slightly, but strongly inhibited C2H2 reduction. Evidence for H2-dependent cytochrome reduction in an H2 uptake-positive strain of R. japonicum bacteroids is presented. In kinetic studies, the rates of reduction of the type b and c cytochromes in the presence of H2 were shown to be severalfold higher than the rates due to endogenous respiration alone. With hydrogenase-deficient mutants of R. japonicum, no measurable effect of H2 on cytochrome reduction was observed. Our results indicate that ubiquinone and cytochromes of types b and c are involved in the oxyhydrogen reaction in R. japonicum.

  4. Diverse Bacteria Affiliated with the Genera Microvirga, Phyllobacterium, and Bradyrhizobium Nodulate Lupinus micranthus Growing in Soils of Northern Tunisia.

    Science.gov (United States)

    Msaddak, Abdelhakim; Durán, David; Rejili, Mokhtar; Mars, Mohamed; Ruiz-Argüeso, Tomás; Imperial, Juan; Palacios, José; Rey, Luis

    2017-03-15

    The genetic diversity of bacterial populations nodulating Lupinus micranthus in five geographical sites from northern Tunisia was examined. Phylogenetic analyses of 50 isolates based on partial sequences of recA and gyrB grouped strains into seven clusters, five of which belong to the genus Bradyrhizobium (28 isolates), one to Phyllobacterium (2 isolates), and one, remarkably, to Microvirga (20 isolates). The largest Bradyrhizobium cluster (17 isolates) grouped with the B. lupini species, and the other five clusters were close to different recently defined Bradyrhizobium species. Isolates close to Microvirga were obtained from nodules of plants from four of the five sites sampled. We carried out an in-depth phylogenetic study with representatives of the seven clusters using sequences from housekeeping genes (rrs, recA, glnII, gyrB, and dnaK) and obtained consistent results. A phylogeny based on the sequence of the symbiotic gene nodC identified four groups, three formed by Bradyrhizobium isolates and one by the Microvirga and Phyllobacterium isolates. Symbiotic behaviors of the representative strains were tested, and some congruence between symbiovars and symbiotic performance was observed. These data indicate a remarkable diversity of L. micranthus root nodule symbionts in northern Tunisia, including strains from the Bradyrhizobiaceae, Methylobacteriaceae, and Phyllobacteriaceae families, in contrast with those of the rhizobial populations nodulating lupines in the Old World, including L. micranthus from other Mediterranean areas, which are nodulated mostly by Bradyrhizobium strains.IMPORTANCELupinus micranthus is a legume broadly distributed in the Mediterranean region and plays an important role in soil fertility and vegetation coverage by fixing nitrogen and solubilizing phosphate in semiarid areas. Direct sowing to extend the distribution of this indigenous legume can contribute to the prevention of soil erosion in pre-Saharan lands of Tunisia. However

  5. Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals.

    Science.gov (United States)

    Zhang, Xin; He, Chuan-Chuan; Liu, Jin-Ming; Li, Hao; Lu, Ke; Fu, Zhi-Qiang; Zhu, Chuan-Gang; Liu, Yi-Ping; Tong, Lai-Bao; Zhou, De-Bao; Zha, Li; Hong, Yang; Jin, Ya-Mei; Lin, Jiao-Jiao

    2017-04-13

    Schistosomiasis japonica is a common zoonosis. Domestic animals are the primary source of infection and play an important role in disease transmission. The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and, for this reason, diagnostics with a higher sensitivity have become increasingly necessary. It was reported that polymerase chain reaction (PCR)-based methods could be used to detect schistosome infection in humans and animals and presented a high sensitivity and specificity. The present study aimed to develop a PCR-based method for detection of Schistosoma japonicum infection in domestic animals. A specific nested-PCR assay was developed to detect S. japonicum infection in domestic animals via amplification of a 231-bp DNA fragment of retrotransposon SjR2. The developed assay was first used in sera and dry blood filter paper (DBFP) from goats and buffaloes at different time points of infection. Then, 78 DBFPs from 39 artificially-infected bovines at 14 and 28 days post-infection and 42 DBFPs from schistosome-negative bovines from the city of Huangshan in the Anhui province were used to evaluate the diagnostic validity. Furthermore, this assay was used to detect S. japonicum infection in domestic animals in Dongzhi and Wangjiang counties. The expected PCR product was detected in eggs and adult worms of S. japonicum and blood samples from S. japonicum-infected goats and water buffaloes, but not from Fasciola and Haemonchus contortus worms. The nested-PCR assay could detect the target S. japonicum DNA in DBFPs from goats and buffaloes after day 3 post-infection. The sensitivity in buffaloes at 14 and 28 days post-infection was 92.30% (36/39) and 100% (39/39), respectively. The specificity was 97.60% (41/42). The positivity rates in Dongzhi and Wangjiang counties were 6.00% and 8.00% in bovines and 22.00% and 16.67% in goats, respectively. The positivity rates in goats in both counties were higher than those

  6. Genome-wide identification of Schistosoma japonicum microRNAs using a deep-sequencing approach.

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    Jian Huang

    Full Text Available BACKGROUND: Human schistosomiasis is one of the most prevalent and serious parasitic diseases worldwide. Schistosoma japonicum is one of important pathogens of this disease. MicroRNAs (miRNAs are a large group of non-coding RNAs that play important roles in regulating gene expression and protein translation in animals. Genome-wide identification of miRNAs in a given organism is a critical step to facilitating our understanding of genome organization, genome biology, evolution, and posttranscriptional regulation. METHODOLOGY/PRINCIPAL FINDINGS: We sequenced two small RNA libraries prepared from different stages of the life cycle of S. japonicum, immature schistosomula and mature pairing adults, through a deep DNA sequencing approach, which yielded approximately 12 million high-quality short sequence reads containing a total of approximately 2 million non-redundant tags. Based on a bioinformatics pipeline, we identified 176 new S. japonicum miRNAs, of which some exhibited a differential pattern of expression between the two stages. Although 21 S. japonicum miRNAs are orthologs of known miRNAs within the metazoans, some nucleotides at many positions of Schistosoma miRNAs, such as miR-8, let-7, miR-10, miR-31, miR-92, miR-124, and miR-125, are indeed significantly distinct from other bilaterian orthologs. In addition, both miR-71 and some miR-2 family members in tandem are found to be clustered in a reversal direction model on two genomic loci, and two pairs of novel S. japonicum miRNAs were derived from sense and antisense DNA strands at the same genomic loci. CONCLUSIONS/SIGNIFICANCE: The collection of S. japonicum miRNAs could be used as a new platform to study the genomic structure, gene regulation and networks, evolutionary processes, development, and host-parasite interactions. Some S. japonicum miRNAs and their clusters could represent the ancestral forms of the conserved orthologues and a model for the genesis of novel miRNAs.

  7. Functions of the Vasa gene in Schistosoma japonicum as assessed by RNA interference.

    Science.gov (United States)

    He, Siyu; Zhu, Lulu; Liu, Fengchun; Liu, Quan; Shao, Yanjing; Hua, Mengqing; Ding, Han; Shao, Wei; Du, Yinan; Hou, Xin; Ren, Cuiping; Liu, Miao; Shen, Jijia

    2018-01-05

    Vasa, an enzyme belonging to the helicase family, contributes to the regulation of reproductive system development in many species. Thus, we hypothesized that the Vasa3 gene may function in the reproductive system of the parasite Schistosoma japonicum (S. japonicum), which is a major causative agent of schistosomiasis. It is a severe disease globally affecting humans and animals. To test this hypothesis, we firstly conducted whole mount in situ hybridization analyses and found that the S. japonicum Vasa3 (SjVasa3) gene was expressed mainly in the reproductive organs. We then explored the reproductive functions of Vasa3 in S. japonicum using RNA interference (RNAi) techniques. Coupled schistosomes collected from mice 28days post infection (dpi) were transfected three times with SjVasa3-specific small interfering RNA (siRNA) and cultured in vitro for up to 10days. As measured by quantitative PCR (qPCR) and Western blot analysis, levels of SjVasa3 mRNA and protein in Vasa siRNA treated worms were significantly reduced compared with untreated and scrambled siRNA treated worms. Confocal laser scanning microscopy (CLSM) images showed markedly siRNA induced changes in the morphology of the reproductive organs, especially in the female ovary, vitellarium and the male testes. SjVasa3 gene silencing also significantly reduced egg production. These data demonstrate that SjVasa3 is essential in reproductive organ development and egg production in S. japonicum, and could be a potential target for developing novel compounds to treat schistosomiasis. Copyright © 2017. Published by Elsevier B.V.

  8. Green synthesis of silver nanoparticles using Ganoderma neo-japonicum Imazeki: a potential cytotoxic agent against breast cancer cells

    National Research Council Canada - National Science Library

    Gurunathan, Sangiliyandi; Raman, Jegadeesh; Abd Malek, Sri Nurestri; John, Priscilla A; Vikineswary, Sabaratnam

    2013-01-01

    .... The aim of our study was to determine the cytotoxic effects of biologically synthesized AgNPs using hot aqueous extracts of the mycelia of Ganoderma neo-japonicum Imazeki on MDA-MB-231 human breast cancer cells...

  9. Evaluation of recombinant fructose-1,6-bisphosphate aldolase ELISA test for the diagnosis of Schistosoma japonicum in water buffaloes.

    Science.gov (United States)

    Peng, Shih-Yi; Lee, Kin-Mu; Tsaihong, John Chin; Cheng, Po-Ching; Fan, Ping-Chin

    2008-12-01

    Fructose-1,6-bisphosphate aldolase (FBPA) is an ubiquitous enzyme essential for glycolysis, gluconeogenesis and the Calvin cycle. It has been demonstrated to induce immune responses and to be useful in the immunodiagnosis of malaria. In this study, FBPA was cloned from the adult worms of Schistosoma japonicum and tested as an antigen for the diagnosis of S. japonicum infection in water buffaloes. Enzyme-linked immunosorbent assay (ELISA) was performed on the sera from 32 infected water buffaloes and 20 negative controls using the recombinant FBPA protein or soluble worm antigen preparation (SWAP) as an antigen. The OD cut-off values were determined to be 0.57 with 100% specificity and 100% sensitivity for the FBPA ELISA and 1.13 with 93.8% specificity and 95.0% sensitivity for the SWAP ELISA. These findings indicate that the recombinant FBPA of S. japonicum should be an useful diagnostic tool for the detection of antibodies against S. japonicum.

  10. Symbiotic effectiveness of Bradyrhizobium ingae in promoting growth of Inga edulis Mart. seedlings

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    Dilacy Sales Porto

    Full Text Available ABSTRACT Inga edulis Mart. is a leguminous tree adapted to acidic and low-fertility soils that establishes symbioses with nitrogen (N-fixing bacteria. The identification of effective bacteria in biological N fixation may bolster the use of I. edulis in degraded or modified areas and agroforestry systems. Therefore, the aims of this study were evaluation of the symbiotic effectiveness of eight strains of the Bradyrhizobium genus native to Roraima in Inga edulis plants, and in vitro evaluation of the ability of the eight strains of Bradyrhizobium to develop plant growth-promoting characteristics. Determination of symbiotic effectiveness was carried out via three experiments: the first in a greenhouse in pots with a sterile substrate; the second in a greenhouse in pots containing non-sterile soil; and the third in a nursery in bags with a non-sterile substrate. Twelve treatments were evaluated: inoculation with eight strains of Bradyrhizobium ingae (ERR 490, ERR 492, ERR 493, ERR 494T, ERR 496, ERR 497, ERR 498, and ERR 569; inoculation with two strains indicated for Inga marginata, BR 6609 and BR 6610 (positive controls; no inoculation but with mineral N; and neither inoculation nor mineral N. All of the experiments were conducted in a completely randomized design with four replicates. The first experiment was conducted for 60 days, and the other experiments were conducted for 100 days. For all of the experiments, the number of nodules, nodule dry matter, root dry matter, shoot dry matter, number of leaflets, plant height, stem diameter, total N in the shoots, root/shoot dry matter ratio, Dickson’s quality index, relative effectiveness, and the Pearson correlation between the variables under study were evaluated. The strains were also evaluated by their ability to solubilize calcium and aluminum phosphates and to produce indolic compounds. The results showed that B. ingae strains were effective in biological N fixation, especially the ERR 493

  11. Assessment of Trichogramma japonicum and T. chilonis as Potential Biological Control Agents of Yellow Stem Borer in Rice

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    Rui Tang

    2017-02-01

    Full Text Available Two species of Trichogramma wasps were assessed for their effectiveness against yellow stem borer Scirpophaga incertulas. A laboratory cage test with T. japonicum and T. chilonis showed that both species parasitized yellow stem borer egg masses at 60.0% ± 9.13% and 40.7% ± 7.11%, respectively, with egg parasitism rates of 15.8% ± 22.2% for T. japonicum and 2.8% ± 5.0% for T. chilonis. Once the host eggs were parasitized, emergence rates were high for both species (95.7% ± 0.12% for T. japonicum and 100% for T. chilonis. In paddy field trials, the two Trichogramma species were released at three densities (50,000/ha, 100,000/ha and 200,000/ha in Southwestern China. Egg mass parasitism was 9% ± 7.7% for T. japonicum and 15% ± 14.1% for T. chilonis, and again only a relatively small fraction of eggs was successfully parasitized. No clear conclusion could be drawn on the most efficient release rate as no significant differences were found among the three release rates. A comparison of field-collected T. japonicum with T. japonicum and T. chilonis mass reared on Corcyra cephalonica showed significantly larger body size and ovipositor length in field-collected wasps, suggesting potentially higher effectiveness on yellow stem borer eggs after at least one generation on the target host. Factors contributing to the low field parasitism rates are discussed.

  12. Assessment of Trichogramma japonicum and T. chilonis as Potential Biological Control Agents of Yellow Stem Borer in Rice.

    Science.gov (United States)

    Tang, Rui; Babendreier, Dirk; Zhang, Feng; Kang, Min; Song, Kai; Hou, Mao-Lin

    2017-02-08

    Two species of Trichogramma wasps were assessed for their effectiveness against yellow stem borer Scirpophaga incertulas. A laboratory cage test with T. japonicum and T. chilonis showed that both species parasitized yellow stem borer egg masses at 60.0% ± 9.13% and 40.7% ± 7.11%, respectively, with egg parasitism rates of 15.8% ± 22.2% for T. japonicum and 2.8% ± 5.0% for T. chilonis. Once the host eggs were parasitized, emergence rates were high for both species (95.7% ± 0.12% for T. japonicum and 100% for T. chilonis). In paddy field trials, the two Trichogramma species were released at three densities (50,000/ha, 100,000/ha and 200,000/ha) in Southwestern China. Egg mass parasitism was 9% ± 7.7% for T. japonicum and 15% ± 14.1% for T. chilonis, and again only a relatively small fraction of eggs was successfully parasitized. No clear conclusion could be drawn on the most efficient release rate as no significant differences were found among the three release rates. A comparison of field-collected T. japonicum with T. japonicum and T. chilonis mass reared on Corcyra cephalonica showed significantly larger body size and ovipositor length in field-collected wasps, suggesting potentially higher effectiveness on yellow stem borer eggs after at least one generation on the target host. Factors contributing to the low field parasitism rates are discussed.

  13. Genetic diversity and symbiotic effectiveness of Bradyrhizobium strains nodulating selected annual grain legumes growing in Ethiopia.

    Science.gov (United States)

    Degefu, Tulu; Wolde-Meskel, Endalkachew; Rasche, Frank

    2017-11-16

    Vigna unguiculata, Vigna radiata and Arachis hypogaea growing in Ethiopia are nodulated by a genetically diverse group of Bradyrhizobium strains. To determine the genetic identity and symbiotic effectiveness of these bacteria, a collection of 36 test strains originating from the root nodules of the three hosts was investigated using multilocus sequence analyses (MLSA) of core genes including 16S rRNA, recA, glnII, gyrB, atpD and dnaK. Sequence analysis of nodA and nifH genes along with tests for symbiotic effectiveness using δ15N analysis were also carried out. The phylogenetic trees derived from the MLSA grouped most test strains into four well-supported distinct positions designated as genospecies I-IV. The maximum likelihood (ML) tree that was constructed based on the nodA gene sequences separated the entire test strains into two lineages, where the majority of the test strains were clustered on one of a well-supported large branch that comprise Bradyrhizobium species from the tropics. This clearly suggested the monophyletic origin of the nodA genes within the bradyrhizobia of tropical origin. The δ15N-based symbiotic effectiveness test of seven selected strains revealed that strains GN100 (δ15N=0.73) and GN102 (δ15N=0.79) were highly effective nitrogen fixers when inoculated to cowpea, thus can be considered as inoculants in cowpea production. It was concluded that Ethiopian soils are a hotspot for rhizobial diversity. This calls for further research to unravel as yet unknown bradyrhizobia nodulating legume host species growing in the country. In this respect, prospective research should also address the mechanisms of symbiotic specificity that could lead to high nitrogen fixation in target legumes.

  14. Respostas antioxidativas em nódulos de caupi inoculado com bradyrhizobium

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    Rayssa Dias Batista

    2016-12-01

    Full Text Available O nitrogênio é considerado elemento essencial para as plantas, pois está presente na composição das mais importantes biomoléculas. As plantas podem obter esse nitrogênio através do processo de fixação biológica pela simbiose entre a planta e as bactérias fixadoras de nitrogênio. A fixação biológica do nitrogênio pode ser afetada por fatores bióticos e abióticos que provocam estresse oxidativo na planta. O sistema antioxidativo enzimático envolve enzimas, como por exemplo dismutase de superóxido (SOD, catalase (CAT, peroxidase de fenóis (POX, que conferem proteção oxidativa às plantas. Desta forma, objetivo deste trabalho foi avaliar o metabolismo antioxidativo em nódulos de plantas de feijão-caupi inoculadas com Bradyrhizobium sp. (estirpe BR 3267. Os resultados mostraram que a inoculação com Bradyrhizobiumsp. foi efetiva e resultou em baixo acúmulo de amônia e diminuição do estresse oxidativo. Concluiu-se que a inoculação de caupi comBradyrhizobium sp. foi efetiva em estimular as enzimas envolvidas no processo oxidativo (SOD, POX e CAT. Estas enzimas foram eficientes na remoção das espécies reativas de oxigênio e promoveram proteção oxidativa para plantas de caupi.

  15. Competition by Bradyrhizobium Strains for Nodulation of the Nonlegume Parasponia andersonii

    Science.gov (United States)

    Trinick, M. J.; Hadobas, P. A.

    1989-01-01

    Bradyrhizobium strains isolated from the nonlegume Parasponia spp. formed a group of strains that were highly competitive for nodulation of P. andersonii when paired with strains isolated from legumes. Strains from legumes, including those of similar effectiveness to NGR231 and CP283, were not able to form nodules as single occupants on P. andersonii in the presence of Parasponia strains. However, NGR86, an isolate from Macroptilium lathyroides, jointly occupied one-third of the nodules formed with each of the three strains isolated from Parasponia spp. Time taken for nodules to appear may have influenced the outcome of competition, since CP283 and all isolates from legumes were slow to nodulate P. andersonii. Among the Parasponia strains, competitiveness for nodulation of P. andersonii was not associated with effectiveness of nitrogen fixation. The highly effective strain CP299 was a poor competitor when paired with the least effective strain NGR231. CP283 was the least competitive of the Parasponia strains but was still able to dominate nodules when paired with legume isolates. Dual occupancy was high, up to 67% when the inoculum contained CP299 and CP273. Both the Muc+ and Muc- types of CP283 form a symbiosis of similar effectiveness and were similarly competitive at high inoculation densities, but the Muc- form was more competitive at low inoculum densities. Both forms frequently occupied the same nodule. Bradyrhizobium strains isolated from Parasponia spp. may have specific genetic information that favor their ability to competitively and effectively infect plants in the genus Parasponia (Ulmaceae) outside the Leguminosae. PMID:16347913

  16. Schistosoma japonicum: An ultraviolet-attenuated cercarial vaccine applicable in the field for water buffaloes

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Y.E.; Jiang, C.F.; Han, J.J.; Li, Y.L.; Ruppel, A. (Tongii Medical Univ., Wuhan, Hubei Province (China))

    1990-07-01

    Water buffaloes were vaccinated three times with 10,000 Schistosoma japonicum cercariae irradiated with ultraviolet (uv) light at a dose of 400 microW x min/cm2. The irradiation was performed with cheap, simple, and portable equipment in a rural area of Hubei Province (People's Republic of China). A challenge infection of 1000 untreated cercariae was given to six vaccinated and six naive control buffaloes, while two vaccinated animals were not challenged. The experiment was terminated 6 weeks after the challenge. Control animals had lost body weight and harbored a mean of 110 worms and 37 eggs per gram of liver. The vaccinated animals gained weight after the challenge and developed 89% resistance to infection with S. japonicum. Since schistosomiasis japonica is nowadays transmitted in China predominantly by domestic livestock, a uv-attenuated cercarial vaccine for bovines may contribute to the control of this disease.

  17. Allelopathy of the invasive plant Bidens frondosa on the seed germination of Geum japonicum var. chinense.

    Science.gov (United States)

    Wang, X F; Hassani, D; Cheng, Z W; Wang, C Y; Wu, J

    2014-12-12

    Five gradient concentrations (0.02, 0.04, 0.06, 0.08, and 0.10 g/mL) of leaching liquors from the roots, stems, and leaves of the invasive plant Bidens frondosa were used as conditioning fluid to examine its influence on seed germination conditions of the native plant Geum japonicum var. chinense in Huangshan. All leaching liquors of organs suppressed the seed germination of Geum japonicum var. chinense and reduced the final germination percentage and rate, and increased the germination inhibition rate, with a bimodal dependence on concentration. The leaching liquor inhibited the seed germination significantly at the concentration of 0.02 g/mL respectively. The seed germination was also inhibited as the concentration reached to 0.04 g/mL and beyond. Hence the allelopathic effects of the organs were significantly enhanced respectively. This phenomenon represented the presence of allelopathy substances in the root, stem and leaf of Bidens frondosa.

  18. Characteristics of IL-17 induction by Schistosoma japonicum infection in C57BL/6 mouse liver

    Science.gov (United States)

    Chen, Dianhui; Luo, Xueping; Xie, Hongyan; Gao, Zhiyan; Fang, Huilong; Huang, Jun

    2013-01-01

    Schistosomiasis japonica is a severe tropical disease caused by the parasitic worm Schistosoma japonicum. Among the most serious pathological effects of S. japonicum infection are hepatic lesions (cirrhosis and fibrosis) and portal hypertension. Interleukin-17 (IL-17) is a pro-inflammatory cytokine involved in the pathogenesis of many inflammatory and infectious conditions, including schistosomiasis. We infected C57BL/6 mice with S. japonicum and isolated lymphocytes from the liver to identify cell subsets with high IL-17 expression and release using flow cytometry and ELISA. Expression and release of IL-17 was significantly higher in hepatic lymphocytes from infected mice compared with control mice in response to both non-specific stimulation with anti-CD3 monoclonal antibody plus/anti-CD28 monoclonal antibody and PMA plus ionomycin. We then compared IL-17 expression in three hepatic T-cell subsets, T helper, natural killer T and γδT cells, to determine the major source of IL-17 during infection. Interleukin-17 was induced in all three subsets by PMA + ionomycin, but γδT lymphocytes exhibited the largest increase in expression. We then established a mouse model to further investigate the role of IL-17 in granulomatous and fibrosing inflammation against parasite eggs. Reducing IL-17 activity using anti-IL-17A antibodies decreased infiltration of inflammatory cells and collagen deposition in the livers of infected C57BL/6 mice. The serum levels of soluble egg antigen (IL) -specific IgGs were enhanced by anti-IL-17A monoclonal antibody blockade, suggesting that IL-17 normally serves to suppress this humoral response. These findings suggest that γδT cells are the most IL-17-producing cells and that IL-17 contributes to granulomatous inflammatory and fibrosing reactions in S. japonicum-infected C57BL/6 mouse liver. PMID:23551262

  19. Repeated Schistosoma japonicum infection following treatment in two cohorts: evidence for host susceptibility to helminthiasis?

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    Elizabeth J Carlton

    Full Text Available In light of multinational efforts to reduce helminthiasis, we evaluated whether there exist high-risk subpopulations for helminth infection. Such individuals are not only at risk of morbidity, but may be important parasite reservoirs and appropriate targets for disease control interventions.We followed two longitudinal cohorts in Sichuan, China to determine whether there exist persistent human reservoirs for the water-borne helminth, Schistosoma japonicum, in areas where treatment is ongoing. Participants were tested for S. japonicum infection at enrollment and two follow-up points. All infections were promptly treated with praziquantel. We estimated the ratio of the observed to expected proportion of the population with two consecutive infections at follow-up. The expected proportion was estimated using a prevalence-based model and, as highly exposed individuals may be most likely to be repeatedly infected, a second model that accounted for exposure using a data adaptive, machine learning algorithm. Using the prevalence-based model, there were 1.5 and 5.8 times more individuals with two consecutive infections than expected in cohorts 1 and 2, respectively (p<0.001 in both cohorts. When we accounted for exposure, the ratio was 1.3 (p = 0.013 and 2.1 (p<0.001 in cohorts 1 and 2, respectively.We found clustering of infections within a limited number of hosts that was not fully explained by host exposure. This suggests some hosts may be particularly susceptible to S. japonicum infection, or that uncured infections persist despite treatment. We propose an explanatory model that suggests that as cercarial exposure declines, so too does the size of the vulnerable subpopulation. In low-prevalence settings, interventions targeting individuals with a history of S. japonicum infection may efficiently advance disease control efforts.

  20. Kerentanan Schistosoma japonicum terhadap Praziquantel di Napu dan Lindu, Sulawesi Tengah Indonesia

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    Anis Nurwidayati

    2016-07-01

    Full Text Available Schistosomiasis in Indonesia are found in Napu, Lindu and Bada highland, Central Sulawesi. This disease was caused by Trematode worm, Schistosoma japonicum. Schistosomiasis still a public health problem which its prevalence increase every year. The large scale treatment by mass chemotherapy using praziquantel was done to reduce the prevalence of schistosomiasis since 1980’s. The objective of this research was to identify the development of resistance in Schistosoma japonicum to praziquantel in endemic areas. Field study was conducted in endemic areas Napu and Lindu in April –November 2011. All of the 80 stool-positive subjects in Napu and 60 stool-positive subjects in Lindu, were treated with a single dose of 60 mg/kg of praziquantel. On three, six, nine, and 12 weeks after treatment, all of the subjects were examined again using the same stool examination. The results showed that on three weeks examination after treatment, stool-negative results were found in all subjects which represents a 100% parasitological cure rate. All stool samples were re-examined six, nine, and 12 weeks after the first treatment and no stool-positive subjects were found. The results indicate that there was no evidence for reduced susceptibility of S.japonicum to praziquantel despite its extensive use in the endemic areas of Napu and Lindu for more than 20 years.

  1. Comparative analysis of codon usage pattern and its influencing factors in Schistosoma japonicum and Ascaris suum.

    Science.gov (United States)

    Mazumder, Gulshana A; Uddin, Arif; Chakraborty, Supriyo

    2017-12-20

    Schistosoma japonicum and Ascaris suum are considered as the major parasites of human which cause various life threatening diseases such as schistomiasis and ascariasis. The codon usage bias (CUB) is known as the phenomenon of more usage of a specific codon than the other synonymous codons for an amino acid. The factors that influence the codon usage bias are mutation pressure, natural selection, gene expression, gene length, GC content, RNA stability, recombination rates, codon position etc. Here we had used various bioinformatic tools and statistical analyses to understand the compositional features, expression level and codon usage bias in the genes of these two species.After estimating the effective number of codon (ENC) in both the species, codon usage bias was found to be low and gene expression was high. The nucleobase A and T were used most often than C and G. From neutrality plot and correspondence analysis it was found that both natural selection and mutation pressure played an important role in shaping the codon usage pattern of both species. Moreover, natural selection played a major role while mutation pressure played a minor role in shaping the codon usage bias in S. japonicum and A.suum. This is the first report on the codon usage biology in S. japonicum and A.suum, and the factors influencing their codon usage bias. These results are expected to be useful for genetic engineering and evolutionary studies.

  2. Cysteine protease inhibitor of Schistosoma japonicum - A parasite-derived negative immunoregulatory factor.

    Science.gov (United States)

    Chen, Lin; He, Baohua; Hou, Wei; He, Li

    2017-03-01

    Studies have shown that cysteine protease inhibitors from some parasites have immunosuppressive effects on the host. We previously have cloned a novel cysteine protease inhibitor from Schistosoma japonicum and purified its recombinant version (protein named rSj-C). Its possible inhibitory effect on the host immune response has not been described.This study shows that rSj-C inhibits lysosomal cysteine protease of murine dendritic cells (DCs). After DCs were incubated with rSj-C and then with soluble adult worm antigen (AWA) of S. japonicum, the mean fluorescence intensity of MHC class II antigens on the surface of DCs decreased significantly by flow cytometry. These results indirectly proved that rSj-C can suppress exogenous-antigen presentation by DCs. The flow cytometric assay revealed that in comparison with control groups, the proportion of CD4(+)CD25(+)Foxp3(+) T cells among CD4(+)CD25(+) T cells of Schistosom-infected mice increased significantly 8 weeks after the infected mice were injected with rSj-C (p ˂ 0.05). Additionally, the expression levels of cytokines IL-4 and TGF-β produced by T cells increased significantly as compared with these levels in the normal group (p ˂ 0.05). These results clearly show that the cysteine protease inhibitor from S. japonicum is a new parasite-derived immunosuppressive factor.

  3. [Killing effect of sodium abietate on adult worms of Schistosoma japonicum in vitro].

    Science.gov (United States)

    Wang, Wen-Bo; Liu, Hong-Jun; Wang, Ben-Jing; Zhou, Xia; Zhang, Jing; Liu, Chen-Chen; Gong, Wei; Zhu-Ge, Hong-Xiang

    2011-06-01

    To observe the killing effect of sodium abietate on adult male and female worms of Schistosoma japonicum in vitro. The mice infected with cercariae of S. japonicum were sacrificed and perfused five weeks later, the adult worms obtained by the portal perfusion method, were cultivated in DMEM medium containing different concentrations of sodium abietate for 3 days, except the controls, then the worms were observed for the death and motility reducing. The worms were stained by hydrochloric acid carmine for the detection of the changes, and the protein of the worms was detected by using the ultraviolet ray-absorption and Bradford method. After the treatment of sodium abietate, the mortality and motility reducing rate of adult worms were higher significantly than the controls; the effect of sodium abietate on male worms was more obvious than on female worms. The male worms' intestinal canal enlarged and appeared black or brown bands or spots after the treatment. The contents of the intestine of female worms were distributed asymmetrically, and the shape of some worms' ovaries was anomalism and the coloring was asymmetrical. Compared with the control group, the protein of adult male and female worms were reduced (P worms of S. japonicum in vitro. It may affect the protein metabolism of the worms.

  4. Taurine drinking ameliorates hepatic granuloma and fibrosis in mice infected with Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    Yan-Rong Yu

    2016-04-01

    Full Text Available In schistosomiasis, egg-induced hepatic granuloma formation is a cytokine-mediated, predominantly CD4+ Th2 immune response that can give rise to hepatic fibrosis. Hepatic fibrosis is the main cause of increased morbidity and mortality in humans with schistosome infection. Taurine has various physiological functions and hepatoprotective properties as well as anti-inflammatory and immunomodulatory activity. However, little is known about the role of taurine in schistosome egg-induced granuloma formation and fibrosis. We aimed to evaluate the therapeutic potential of taurine as preventative treatment for Schistosoma japonicum infection. Mice infected with S. japonicum cercariae were supplied with taurine drinking water (1% w/v for 4 weeks starting at 4 weeks post-infection. Taurine supplementation significantly improved the liver pathologic findings, reduced the serum levels of aminotransferases and area of hepatic granuloma, and prevented fibrosis progression. In addition, taurine decreased the expression of the granulomatous and fibrogenic mediators transforming growth factor β1, tumor necrosis factor α, monocyte chemotactic protein 1α and macrophage inflammatory protein 1α as well as the endoplasmic reticulum stress marker glucose-regulated protein 78. Thus, taurine can significantly attenuate S. japonicum egg-induced hepatic granuloma and fibrosis, which may depend in part on the downregulation of some relevant cytokine/chemokines and reducing the endoplasmic reticulum stress response.

  5. High prevalence of Schistosoma japonicum and Fasciola gigantica in bovines from Northern Samar, the Philippines.

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    Catherine A Gordon

    2015-02-01

    Full Text Available The cause of zoonotic schistosomiasis in the Philippines is Schistosoma japonicum, which infects up to 46 mammalian hosts, including humans and bovines. In China, water buffaloes have been identified as major reservoir hosts for schistosomiasis japonica, contributing up to 75% of human transmission. In the Philippines, water buffaloes (carabao; Bubalus bubalis carabanesis have, historically, been considered unimportant reservoirs. We therefore revisited the possible role of bovines in schistosome transmission in the Philippines, using the recently described formalin-ethyl acetate sedimentation (FEA-SD technique and a qPCR assay to examine fecal samples from 153 bovines (both carabao and cattle from six barangays in Northern Samar. A high prevalence of S. japonicum was found using qPCR and FEA-SD in both cattle (87.50% and 77.08%, respectively and carabao (80.00% and 55.24%, respectively. The average daily egg output for each bovine was calculated at 195,000. High prevalence and infection intensity of F. gigantica was also found in the bovines by qPCR and FEA-SD (95.33% and 96.00%, respectively. The identification of bovines as major reservoir hosts for S. japonicum transmission suggests that bovine treatment and/or vaccination, as one becomes available, should be included in any future control program that aims to reduce the disease burden due to schistosomiasis in the Philippines.

  6. New Perspectives on Host-Parasite Interplay by Comparative Transcriptomic and Proteomic Analyses of Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    2006-04-01

    Full Text Available Schistosomiasis remains a serious public health problem with an estimated 200 million people infected in 76 countries. Here we isolated ~ 8,400 potential protein-encoding cDNA contigs from Schistosoma japonicum after sequencing circa 84,000 expressed sequence tags. In tandem, we undertook a high-throughput proteomics approach to characterize the protein expression profiles of a number of developmental stages (cercariae, hepatic schistosomula, female and male adults, eggs, and miracidia and tissues at the host-parasite interface (eggshell and tegument by interrogating the protein database deduced from the contigs. Comparative analysis of these transcriptomic and proteomic data, the latter including 3,260 proteins with putative identities, revealed differential expression of genes among the various developmental stages and sexes of S. japonicum and localization of putative secretory and membrane antigens, enzymes, and other gene products on the adult tegument and eggshell, many of which displayed genetic polymorphisms. Numerous S. japonicum genes exhibited high levels of identity with those of their mammalian hosts, whereas many others appeared to be conserved only across the genus Schistosoma or Phylum Platyhelminthes. These findings are expected to provide new insights into the pathophysiology of schistosomiasis and for the development of improved interventions for disease control and will facilitate a more fundamental understanding of schistosome biology, evolution, and the host-parasite interplay.

  7. Taurine drinking ameliorates hepatic granuloma and fibrosis in mice infected with Schistosoma japonicum.

    Science.gov (United States)

    Yu, Yan-Rong; Ni, Xian-Qiang; Huang, Jie; Zhu, Yong-Hong; Qi, Yong-Fen

    2016-04-01

    In schistosomiasis, egg-induced hepatic granuloma formation is a cytokine-mediated, predominantly CD4(+) Th2 immune response that can give rise to hepatic fibrosis. Hepatic fibrosis is the main cause of increased morbidity and mortality in humans with schistosome infection. Taurine has various physiological functions and hepatoprotective properties as well as anti-inflammatory and immunomodulatory activity. However, little is known about the role of taurine in schistosome egg-induced granuloma formation and fibrosis. We aimed to evaluate the therapeutic potential of taurine as preventative treatment for Schistosoma japonicum infection. Mice infected with S. japonicum cercariae were supplied with taurine drinking water (1% w/v) for 4 weeks starting at 4 weeks post-infection. Taurine supplementation significantly improved the liver pathologic findings, reduced the serum levels of aminotransferases and area of hepatic granuloma, and prevented fibrosis progression. In addition, taurine decreased the expression of the granulomatous and fibrogenic mediators transforming growth factor β1, tumor necrosis factor α, monocyte chemotactic protein 1α and macrophage inflammatory protein 1α as well as the endoplasmic reticulum stress marker glucose-regulated protein 78. Thus, taurine can significantly attenuate S. japonicum egg-induced hepatic granuloma and fibrosis, which may depend in part on the downregulation of some relevant cytokine/chemokines and reducing the endoplasmic reticulum stress response.

  8. New perspectives on host-parasite interplay by comparative transcriptomic and proteomic analyses of Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Feng Liu

    2006-04-01

    Full Text Available Schistosomiasis remains a serious public health problem with an estimated 200 million people infected in 76 countries. Here we isolated ~ 8,400 potential protein-encoding cDNA contigs from Schistosoma japonicum after sequencing circa 84,000 expressed sequence tags. In tandem, we undertook a high-throughput proteomics approach to characterize the protein expression profiles of a number of developmental stages (cercariae, hepatic schistosomula, female and male adults, eggs, and miracidia and tissues at the host-parasite interface (eggshell and tegument by interrogating the protein database deduced from the contigs. Comparative analysis of these transcriptomic and proteomic data, the latter including 3,260 proteins with putative identities, revealed differential expression of genes among the various developmental stages and sexes of S. japonicum and localization of putative secretory and membrane antigens, enzymes, and other gene products on the adult tegument and eggshell, many of which displayed genetic polymorphisms. Numerous S. japonicum genes exhibited high levels of identity with those of their mammalian hosts, whereas many others appeared to be conserved only across the genus Schistosoma or Phylum Platyhelminthes. These findings are expected to provide new insights into the pathophysiology of schistosomiasis and for the development of improved interventions for disease control and will facilitate a more fundamental understanding of schistosome biology, evolution, and the host-parasite interplay.

  9. Closely related strains of Bradyrhizobium contained in commercial inoculates of soybean are identified by a set of PCR reactions

    OpenAIRE

    López, Silvina M. Y.; Balatti, Pedro Alberto

    2012-01-01

    The aim of this work was to identify closely related rhizobia, used to formulate commercial inoculants, using polymerase chain reaction (PCR). Repetitive extra-genic palindromic (REP) and BOX fingerprints hardly discriminate among a set of commercial strains. PCR targeted at repetitive RSα successfully allow discriminating within representatives of Bradyrhizobium. These fingerprints clustered isolates at a higher level of similarity and proved to be an important tool to complement the informa...

  10. Bradyrhizobium viridifuturi sp. nov., encompassing nitrogen-fixing symbionts of legumes used for green manure and environmental services.

    Science.gov (United States)

    Helene, Luisa Caroline Ferraz; Delamuta, Jakeline Renata Marçon; Ribeiro, Renan Augusto; Ormeño-Orrillo, Ernesto; Rogel, Marco Antonio; Martínez-Romero, Esperanza; Hungria, Mariangela

    2015-12-01

    Symbiotic nitrogen-fixing bacteria, commonly called rhizobia, are agronomically important because they can provide significant amounts of nitrogen to plants and help in recovery of impoverished soils and improvement of degraded environments. In recent years, with advances in molecular techniques, several studies have shown that these bacteria have high levels of genetic diversity, resulting in taxonomic reclassifications and descriptions of new species. However, despite the advances achieved, highly conserved 16S ribosomal genes (16S rRNA) do not elucidate differences between species of several genera, including the genus Bradyrhizobium. Other methodologies, such as multilocus sequence analysis (MLSA), have been used in such cases, with good results. In this study, three strains (SEMIAs 690T, 6387 and 6428) of the genus Bradyrhizobium, isolated from nitrogen-fixing nodules of Centrosema and Acacia species, without clear taxonomic positions, were studied. These strains differed from genetically closely related species according to the results of MLSA of four housekeeping genes (dnaK, glnII, gyrB and recA) and nucleotide identities of the concatenated genes with those of related species ranged from 87.8 % to 95.7 %, being highest with Bradyrhizobium elkanii. DNA-DNA hybridization (less than 32 % DNA relatedness) and average nucleotide identity values of the whole genomes (less than 90.5 %) indicated that these strains represented a novel species, and phenotypic traits were determined. Our data supported the description of the SEMIA strains as Bradyrhizobium viridifuturi sp. nov., and SEMIA 690T ( = CNPSo 991T = C 100aT = BR 1804T = LMG 28866T), isolated from Centrosema pubescens, was chosen as type strain.

  11. EFFECT OF Nmin CONTENT ON NODULATION IN YELLOW LUPIN (Lupinus luteus L. IN THE PRESENCE OF Bradyrhizobium lupini AND GENISTEIN

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    Janusz PRUSIŃSKI

    2014-03-01

    Full Text Available A two-factor pot experiment was carried out in the years 2009-2011 under a permanent plastic-covered tunnel greenhouse at the Mochełek Experimental Station of the University of Technology and Life Sciences in Bydgoszcz (POLAND. The subject of research was the traditional cultivar of yellow lupin, Mister. The design of the experiment included 5 levels of Nmin content in the plant growth medium: from 4.45 mg Nmin.kg-1.d.w.– very low content to 22.5 mg Nmin.kg-1d.w. – very high content. Seeds of yellow lupin were inoculated with agar inoculum containing Bradyrhizobium lupini with or without the addition of genistein. Along with the plant growth, gradual decrease in the mean content of Nmin was observed, especially in the nitrogenous form until the stage of full bloom of lupin plants. No significant effect of the Nmin content was found on the initiation of the nodulation process, number and dry weight of nodules, independently of seed inoculation with Bradyrhizobium lupini and application of genistein. Average or higher content of Nmin in the medium was the reason of the lack of survival in symbiotic bacteria Bradyrhizobium lupini capable of nodulation in the medium after lupin harvest. Only in the case of low and very low Nmin content in the medium, the addition of genistein to bacterial inoculum did affect the increase in the survival of symbiotic bacteria after lupin harvest.

  12. Photosynthesis, nitrogen allocation and specific leaf area in invasive Eupatorium adenophorum and native Eupatorium japonicum grown at different irradiances.

    Science.gov (United States)

    Feng, Yu-Long

    2008-06-01

    The mechanisms underlying biological invasions are still not well elucidated. In this study, ecophysiological traits of invasive Eupatorium adenophorum and native E. japonicum were compared at 10 irradiances in field. I hypothesized that the invader may allocate a higher fraction of leaf nitrogen (N) to photosynthesis and have higher light-saturated photosynthetic rate (P(max)) and specific leaf area (SLA) than E. japonicum. The invader had a significantly higher ability to acclimate to high irradiance than E. japonicum, while it showed a similar shade-tolerant ability. The invader indeed allocated a higher fraction of leaf N to photosynthesis than E. japonicum, which, with its high leaf N content (N(A)), resulted in a higher N content in photosynthesis (N(P)), contributing to its higher biochemical capacity for photosynthesis and P(max). However, the invader had a significantly lower SLA than E. japonicum, contributing to its higher P(max) but increasing its area-based leaf construction cost. The abilities to acclimate to a wider range of irradiance and to allocate a higher fraction of leaf N to photosynthesis, and the higher P(max), N(A), N(P) and leaf area ratio may contribute to the invasion of the invader. High SLA is not always necessary for invasive species.

  13. NodD1 and NodD2 Are Not Required for the Symbiotic Interaction of Bradyrhizobium ORS285 with Nod-Factor-Independent Aeschynomene Legumes.

    Science.gov (United States)

    Nouwen, Nico; Fardoux, Joel; Giraud, Eric

    2016-01-01

    Photosynthetic Bradyrhizobium strain ORS285 forms nitrogen-fixing nodules on the roots and stems of tropical aquatic legumes of the Aeschynomene genus. Depending on the Aeschynomene species, this symbiotic interaction does or does not rely on the synthesis of Nod-factors (NFs). However, whether during the interaction of Bradyrhizobium ORS285 with NF-independent Aeschynomene species the nod genes are expressed and if the general regulator NodD plays a symbiotic role is unknown. Expression studies showed that in contrast to the interaction with the NF-dependent Aeschynomene species, A. afraspera, the Bradyrhizobium ORS285 nod genes are not induced upon contact with the NF-independent host plant A. indica. Mutational analysis of the two nodD genes present in ORS285, showed that deletion of nodD1 and nodD2 did not affect the symbiotic interaction between Bradyrhizobium ORS285 and A. indica whereas the deletions had an effect on the symbiotic interaction with A. afraspera plants. In addition, when the expression of nod genes was artificially induced by adding naringenin to the plant growth medium, the nodulation of A. indica by Bradyrhizobium ORS285 is delayed and resulted in lower nodule numbers.

  14. Novel European free-living, non-diazotrophic Bradyrhizobium isolates from contrasting soils that lack nodulation and nitrogen fixation genes – a genome comparison

    Science.gov (United States)

    Jones, Frances Patricia; Clark, Ian M.; King, Robert; Shaw, Liz J.; Woodward, Martin J.; Hirsch, Penny R.

    2016-01-01

    The slow-growing genus Bradyrhizobium is biologically important in soils, with different representatives found to perform a range of biochemical functions including photosynthesis, induction of root nodules and symbiotic nitrogen fixation and denitrification. Consequently, the role of the genus in soil ecology and biogeochemical transformations is of agricultural and environmental significance. Some isolates of Bradyrhizobium have been shown to be non-symbiotic and do not possess the ability to form nodules. Here we present the genome and gene annotations of two such free-living Bradyrhizobium isolates, named G22 and BF49, from soils with differing long-term management regimes (grassland and bare fallow respectively) in addition to carbon metabolism analysis. These Bradyrhizobium isolates are the first to be isolated and sequenced from European soil and are the first free-living Bradyrhizobium isolates, lacking both nodulation and nitrogen fixation genes, to have their genomes sequenced and assembled from cultured samples. The G22 and BF49 genomes are distinctly different with respect to size and number of genes; the grassland isolate also contains a plasmid. There are also a number of functional differences between these isolates and other published genomes, suggesting that this ubiquitous genus is extremely heterogeneous and has roles within the community not including symbiotic nitrogen fixation. PMID:27162150

  15. Novel European free-living, non-diazotrophic Bradyrhizobium isolates from contrasting soils that lack nodulation and nitrogen fixation genes - a genome comparison

    Science.gov (United States)

    Jones, Frances Patricia; Clark, Ian M.; King, Robert; Shaw, Liz J.; Woodward, Martin J.; Hirsch, Penny R.

    2016-05-01

    The slow-growing genus Bradyrhizobium is biologically important in soils, with different representatives found to perform a range of biochemical functions including photosynthesis, induction of root nodules and symbiotic nitrogen fixation and denitrification. Consequently, the role of the genus in soil ecology and biogeochemical transformations is of agricultural and environmental significance. Some isolates of Bradyrhizobium have been shown to be non-symbiotic and do not possess the ability to form nodules. Here we present the genome and gene annotations of two such free-living Bradyrhizobium isolates, named G22 and BF49, from soils with differing long-term management regimes (grassland and bare fallow respectively) in addition to carbon metabolism analysis. These Bradyrhizobium isolates are the first to be isolated and sequenced from European soil and are the first free-living Bradyrhizobium isolates, lacking both nodulation and nitrogen fixation genes, to have their genomes sequenced and assembled from cultured samples. The G22 and BF49 genomes are distinctly different with respect to size and number of genes; the grassland isolate also contains a plasmid. There are also a number of functional differences between these isolates and other published genomes, suggesting that this ubiquitous genus is extremely heterogeneous and has roles within the community not including symbiotic nitrogen fixation.

  16. ORF Alignment: NC_004463 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ator ... [Bradyrhizobium japonicum USDA 110] ... Length = 225 ... Query: 42 ... ILIVEDDQETRNLVARYLRENSFNVGMAANGREMDR...YISQNRVDLIVLDLMLPGEDGLSL 101 ... ILIVEDDQETRNLVARYLRENSFNVGMAANGREMDR...YISQNRVDLIVLDLMLPGEDGLSL Sbjct: 1 ... ILIVEDDQETRNLVARYLRENSFNVGMAANGREMDRYISQNRVDLIVLDLMLPGEDGLSL

  17. ORF Alignment: NC_004463 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available y protein ... [Bradyrhizobium japonicum USDA 110] ... Length = 108 ... Query: 208 DIAILHRASRFIDQN...LGRNDLSVATICREIGVSRSVLYRVFAPLAGVADYIRGRRLEAIH 267 ... DIAILHRASRFIDQNLGRND...LSVATICREIGVSRSVLYRVFAPLAGVADYIRGRRLEAIH Sbjct: 1 ... DIAILHRASRFIDQNLGRNDLSVATICREIGVSRSVLYRVFAPLAGVADYIRGRRLEAIH 60 ...

  18. ORF Alignment: NC_004463 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available d sensor ... and regulator [Bradyrhizobium japonicum USDA 110] ... Length = 79 ... Query: 306 MAEQLARRSDYIATF...SAHLTHELKSPLTSIKGAAELLQDSVQGKSEDLTPAEQKTFIANI 365 ... MAEQLARRSDYIATF...SAHLTHELKSPLTSIKGAAELLQDSVQGKSEDLTPAEQKTFIANI Sbjct: 1 ... MAEQLARRSDYIATFSAHLTHELKSPLTSIKGAAELLQDSVQGKSEDLTPAEQKTFIANI 60 ...

  19. The effect of colostrum on pigs pre-natally or post-natally exposed to Schistosoma japonicum

    DEFF Research Database (Denmark)

    Techau, M.E.; Johansen, M.V.; Lind, Peter

    2004-01-01

    A responses, in groups of pigs pre-natally, pre-natally + post-natally or post-natally exposed to S. japonicum. Results suggest that pre-natal exposure and immune colostrum did not affect the establishment of a post-natal challenge infection. However, immune colostrum seemed to increase the levels of septal......Pre-natal infection of Schistosoma japonicum in pigs may prove to be a useful model in shedding light on human pre-natal schistosomiasis. This study describes the effects of immune colostrum on worm burdens, tissue egg counts, liver pathology and crude worm or egg antigen-specific IgG and Ig...... fibrosis in pre-natally exposed pigs. These findings indicate that further investigations will prove valuable, elucidating the influence of the parasitological and immunological status of the sow, on pre-natally exposed pigs, and on the ability of these pigs to develop resistance against S. japonicum later...

  20. Molecular differentiation of Schistosoma japonicum and Schistosoma mekongi by real-time PCR with high resolution melting analysis.

    Science.gov (United States)

    Kongklieng, Amornmas; Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Lulitanond, Viraphong; Sri-Aroon, Pusadee; Limpanont, Yanin; Maleewong, Wanchai

    2013-12-01

    Human schistosomiasis caused by Schistosoma japonicum and Schistosoma mekongi is a chronic and debilitating helminthic disease still prevalent in several countries of Asia. Due to morphological similarities of cercariae and eggs of these 2 species, microscopic differentiation is difficult. High resolution melting (HRM) real-time PCR is developed as an alternative tool for the detection and differentiation of these 2 species. A primer pair was designed for targeting the 18S ribosomal RNA gene to generate PCR products of 156 base pairs for both species. The melting points of S. japonicum and S. mekongi PCR products were 84.5±0.07℃ and 85.7±0.07℃, respectively. The method permits amplification from a single cercaria or an egg. The HRM real-time PCR is a rapid and simple tool for differentiation of S. japonicum and S. mekongi in the intermediate and final hosts.

  1. Survey of traditional Dai medicine reveals species confusion and potential safety concerns: a case study on Radix Clerodendri Japonicum.

    Science.gov (United States)

    Duan, Bao-Zhong; Fang, Hai-Lan; Li, Xi-Wen; Huang, Lin-Fang; Ping, Wang; Chen, Shi-Lin

    2017-06-01

    The adulteration of herbal products is a threat to consumer safety. In the present study, we surveyed the species composition of commercial Radix Clerodendri Japonicum products using DNA barcoding as a supervisory method. A reference database for plant-material DNA-barcode was successfully constructed with 48 voucher samples from 12 Clerodendrum species. The database was used to identify 27 Radix Clerodendri Japonicum decoction piece samples purchased from drug stores and hospitals. The DNA sequencing results revealed that only 1 decoction piece (3.70%) was authentic C. japonicum, as recorded in the Dai Pharmacopeia, whereas the other samples were all adulterants, indicating a potential safety issue. The results indicate that decoction pieces that are available in the market have complex origins and that DNA barcoding is a suitable tool for regulation of Dai medicines. Copyright © 2017 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

  2. Floral organ MADS-box genes in Cercidiphyllum japonicum (Cercidiphyllaceae): Implications for systematic evolution and bracts definition.

    Science.gov (United States)

    Jin, Yupei; Wang, Yubing; Zhang, Dechun; Shen, Xiangling; Liu, Wen; Chen, Faju

    2017-01-01

    The dioecious relic Cercidiphyllum japonicum is one of two species of the sole genus Cercidiphyllum, with a tight inflorescence lacking an apparent perianth structure. In addition, its systematic place has been much debated and, so far researches have mainly focused on its morphology and chloroplast genes. In our investigation, we identified 10 floral organ identity genes, including four A-class, three B-class, two C-class and one D-class. Phylogenetic analyses showed that all ten genes are grouped with Saxifragales plants, which confirmed the phylogenetic place of C. japonicum. Expression patterns of those genes were examined by quantitative reverse transcriptase PCR, with some variations that did not completely coincide with the ABCDE model, suggesting some subfunctionalization. As well, our research supported the idea that thebract actually is perianth according to our morphological and molecular analyses in Cercidiphyllum japonicum.

  3. Dynamics of Th17 cells and their role in Schistosoma japonicum infection in C57BL/6 mice.

    Directory of Open Access Journals (Sweden)

    Xiaoyun Wen

    2011-11-01

    Full Text Available BACKGROUND: The current knowledge of immunological responses to schistosomiasis, a major tropical helminthic disease, is insufficient, and a better understanding of these responses would support vaccine development or therapies to control granuloma-associated immunopathology. CD4(+ T cells play critical roles in both host immune responses against parasitic infection and immunopathology in schistosomiasis. The induction of T helper (Th1, Th2 and T regulatory (Treg cells and their roles in schistosome infections are well-illustrated. However, little in vivo data are available on the dynamics of Th17 cells, another important CD4(+ T cell subset, after Schistosoma japonicum infection or whether these cells and their defining IL-17 cytokine mediate host protective responses early in infection. METHODOLOGY: Levels of Th17 and the other three CD4(+ T cell subpopulations and the cytokines related to induction or repression of Th17 cell generation in different stages of S. japonicum infection were observed. Contrary to reported in vitro studies, our results showed that the Th17 cells were induced along with the Th1, Th2, Treg cells and the IFN-γ and IL-4 cytokines in S. japonicum infected mice. The results also suggested that S. japonicum egg antigens but not adult worm antigens preferentially induced Th17 cell generation. Furthermore, decreasing IL-17 with a neutralizing anti-IL-17 monoclonal antibody (mAb increased schistosome-specific antibody levels and partial protection against S. japonicum infection in mice. CONCLUSIONS: Our study is the first to report the dynamics of Th17 cells during S. japonicum infection and indicate that Th17 cell differentiation results from the integrated impact of inducing and suppressive factors promoted by the parasite. Importantly, our findings suggest that lower IL-17 levels may result in favorable host protective responses. This study significantly contributes to the understanding of immunity to schistosomiasis and

  4. Dynamics of Th17 cells and their role in Schistosoma japonicum infection in C57BL/6 mice.

    Science.gov (United States)

    Wen, Xiaoyun; He, Lei; Chi, Ying; Zhou, Sha; Hoellwarth, Jason; Zhang, Cui; Zhu, Jifeng; Wu, Calvin; Dhesi, Shawn; Wang, Xuefeng; Liu, Feng; Su, Chuan

    2011-11-01

    The current knowledge of immunological responses to schistosomiasis, a major tropical helminthic disease, is insufficient, and a better understanding of these responses would support vaccine development or therapies to control granuloma-associated immunopathology. CD4(+) T cells play critical roles in both host immune responses against parasitic infection and immunopathology in schistosomiasis. The induction of T helper (Th)1, Th2 and T regulatory (Treg) cells and their roles in schistosome infections are well-illustrated. However, little in vivo data are available on the dynamics of Th17 cells, another important CD4(+) T cell subset, after Schistosoma japonicum infection or whether these cells and their defining IL-17 cytokine mediate host protective responses early in infection. Levels of Th17 and the other three CD4(+) T cell subpopulations and the cytokines related to induction or repression of Th17 cell generation in different stages of S. japonicum infection were observed. Contrary to reported in vitro studies, our results showed that the Th17 cells were induced along with the Th1, Th2, Treg cells and the IFN-γ and IL-4 cytokines in S. japonicum infected mice. The results also suggested that S. japonicum egg antigens but not adult worm antigens preferentially induced Th17 cell generation. Furthermore, decreasing IL-17 with a neutralizing anti-IL-17 monoclonal antibody (mAb) increased schistosome-specific antibody levels and partial protection against S. japonicum infection in mice. Our study is the first to report the dynamics of Th17 cells during S. japonicum infection and indicate that Th17 cell differentiation results from the integrated impact of inducing and suppressive factors promoted by the parasite. Importantly, our findings suggest that lower IL-17 levels may result in favorable host protective responses. This study significantly contributes to the understanding of immunity to schistosomiasis and may aid in developing interventions to protect hosts

  5. Interrelationships between Bacillus sp. CHEP5 and Bradyrhizobium sp. SEMIA6144 in the induced systemic resistance against Sclerotium rolfsii and symbiosis on peanut plants.

    Science.gov (United States)

    Figueredo, Maria Soledad; Tonelli, Maria Laura; Taurian, Tania; Angelini, Jorge; Ibanez, Fernando; Valetti, Lucio; Munoz, Vanina; Anzuay, Maria Soledad; Luduena, Liliana; Fabra, Adriana

    2014-12-01

    Plant-growth-promoting bacteria are often used to enhance crop yield and for biological control of phytopathogens. Bacillus sp. CHEP5 is a biocontrol agent that induces systemic resistance (ISR) in Arachis hypogaea L. (peanut) against Sclerotium rolfsii, the causal agent of root and stem wilt. In this work, the effect of the co-inoculation of Bacillus sp. CHEP5 and the peanut nodulating strain Bradyrhizobium sp. SEMIA 6144 was studied on induction of both systemic resistance and nodulation processes. Bradyrhizobium sp. SEMIA 6144 did not affect the ability of Bacillus sp. CHEP5 to protect peanut plants from S. rolfsii by ISR and the priming in challenged-plants, as evidenced by an increment in phenylalanine ammonia-lyase enzyme activity. Additionally, the capacity of Bradyrhizobium sp. SEMIA 6144 to induce nodule formation in pathogen-challenged plants was improved by the presence of Bacillus sp. CHEP5.

  6. Bradyrhizobium Populations Occur in Deep Soil under the Leguminous Tree Acacia albida.

    Science.gov (United States)

    Dupuy, N C; Dreyfus, B L

    1992-08-01

    Soil cores were drilled under the leguminous tree Acacia albida growing in two different ecoclimatic zones of West Africa: the Sahelian area (100 to 500 mm of annual rainfall) and the Sudano-Guinean area (1,000 to 1,500 mm of annual rainfall). Soil samples were collected at different depths from the surface down to the water table level and analyzed for the presence of rhizobia able to nodulate A. albida. In both areas, population densities of rhizobia were substantially greater near the water table than near the surface. In the Sahelian area, rhizobia were present as deep as 34 m at a concentration of 1.3 x 10/g of soil. In the Sudano-Guinean area, population densities at 0.5 to 4.5 m depth were higher than in the Sahelian area and, at several depths, comparable to that of temperate soils supporting legume crops (10 rhizobia per g of soil). Surface and deep soil isolates from all four sites were found to be slow-growing rhizobia (Bradyrhizobium sp.). The proportion of effective isolates was almost the same within surface and deep soils.

  7. Lupinus albus plants acquire mercury tolerance when inoculated with an Hg-resistant Bradyrhizobium strain.

    Science.gov (United States)

    Quiñones, Miguel A; Ruiz-Díez, Beatriz; Fajardo, Susana; López-Berdonces, Miguel A; Higueras, Pablo L; Fernández-Pascual, Mercedes

    2013-12-01

    One strain of Bradyrhizobium canariense (L-7AH) was selected for its metal-resistance and ability to nodulate white lupin (Lupinus albus L.) plants, from a collection of rhizobial strains previously created from soils of the Almadén mining district (Spain) with varying levels of Hg contamination. Plants were inoculated with either strain L-7AH (Hg-tolerant) or L-3 (Hg-sensitive, used as control), and watered with nutrient solutions supplemented with various concentrations (0-200 μM) of HgCl2 in a growth chamber. L. albus inoculated with L-7AH were able to nodulate even at the highest concentration of Hg while those inoculated with L-3 had virtually no nodules at Hg concentrations above 25 μM. Plants inoculated with L-7AH, but not those with the control strain, were able to accumulate large amounts of Hg in their roots and nodules. Nodulation with L-7AH allowed plants to maintain constant levels of both chlorophylls and carotenoids in their leaves and a high photosynthetic efficiency, whereas in those inoculated with L-3 both pigment content and photosynthetic efficiency decreased significantly as Hg concentration increased. Nitrogenase activity of plants nodulated with L-7AH remained fairly constant at all concentrations of Hg used. Results suggest that this symbiotic pair may be used for rhizoremediation of Hg-contaminated soils. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  8. Pongamia pinnata inoculated with Bradyrhizobium liaoningense PZHK1 shows potential for phytoremediation of mine tailings.

    Science.gov (United States)

    Yu, Xiumei; Li, Yangxin; Li, Yanmei; Xu, Chaohua; Cui, Yongliang; Xiang, Quanju; Gu, Yunfu; Zhao, Ke; Zhang, Xiaoping; Penttinen, Petri; Chen, Qiang

    2017-02-01

    Mine tailings contain high concentrations of metal contaminants and only little nutrients, making the tailings barren for decades after the mining has been terminated. Effective phytoremediation of mine tailings calls for deep-rooted, metal accumulating, and soil fertility increasing plants with tolerance against harsh environmental conditions. We assessed the potential of the biofuel leguminous tree Pongamia pinnata inoculated with plant growth promoting rhizobia to remediate iron-vanadium-titanium oxide (V-Ti magnetite) mine tailing soil by pot experiment and in situ remediation test. A metal tolerant rhizobia strain PZHK1 was isolated from the tailing soil and identified as Bradyrhizobium liaoningense by phylogenetic analysis. Inoculation with PZHK1 increased the growth of P. pinnata both in V-Ti magnetite mine tailings and in Ni-contaminated soil. Furthermore, inoculation increased the metal accumulation capacity and superoxide dismutase activity of P. pinnata. The concentrations of Ni accumulated by inoculated plants were higher than the hyperaccumulator threshold. Inoculated P. pinnata accumulated high concentration of Fe, far exceeding the upper limit (1000 mg kg-1) of Fe in plant tissue. In summary, P. pinnata-B. liaoningense PZHK1 symbiosis showed potential to be applied as an effective phytoremediation technology for mine tailings and to produce biofuel feedstock on the marginal land.

  9. Identification of Bradyrhizobium elkanii Genes Involved in Incompatibility with Vigna radiata

    Directory of Open Access Journals (Sweden)

    Hien P. Nguyen

    2017-12-01

    Full Text Available The establishment of a root nodule symbiosis between a leguminous plant and a rhizobium requires complex molecular interactions between the two partners. Compatible interactions lead to the formation of nitrogen-fixing nodules, however, some legumes exhibit incompatibility with specific rhizobial strains and restrict nodulation by the strains. Bradyrhizobium elkanii USDA61 is incompatible with mung bean (Vigna radiata cv. KPS1 and soybean cultivars carrying the Rj4 allele. Here, we explored genetic loci in USDA61 that determine incompatibility with V. radiata KPS1. We identified five novel B. elkanii genes that contribute to this incompatibility. Four of these genes also control incompatibility with soybean cultivars carrying the Rj4 allele, suggesting that a common mechanism underlies nodulation restriction in both legumes. The fifth gene encodes a hypothetical protein that contains a tts box in its promoter region. The tts box is conserved in genes encoding the type III secretion system (T3SS, which is known for its delivery of virulence effectors by pathogenic bacteria. These findings revealed both common and unique genes that are involved in the incompatibility of B. elkanii with mung bean and soybean. Of particular interest is the novel T3SS-related gene, which causes incompatibility specifically with mung bean cv. KPS1.

  10. Pengaruh ketiadaan inang terhadap tanggap reproduksi Trichogrammatoidea armigera Nagaraja dan Trichogramma japonicum Ashmed (Hymenoptera: Trichogrammatoidea) dan implikasinya terhadap penerimaan inang

    OpenAIRE

    Anis Rohmani; Damayanti Buchori; Adha Sari

    2017-01-01

    Trichogrammatoidea armigera and Trichogramma japonicum are polyphagous egg parastioids, that are important as natural enemies. The objective of this research was to study the effect of host deprivation on reproductive capacity of T. armigera and T. japonicum. This study consists of 8 treatments, host deprivation : 0 hour, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, and 48 hours. Host were replaced every 24 hours. Results showed there are difference with respect of how the two parasitoids res...

  11. Efeito do tratamento das sementes de soja com fungicidas e período de armazenamento na resposta da planta inoculada com Bradyrhizobium. = Effect of treating seeds with fungicides and storage on the response of plants inoculated with Bradyrhizobium.

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Pereira

    2010-12-01

    Full Text Available O tratamento químico e a inoculação das sementes com bactérias do gênero Bradyrhizobium são práticas rotineiras na cultura da soja, entretanto a interação entre estas duas práticas pode prejudicar a fixação biológica de nitrogênio e comprometer o crescimento e desenvolvimento das plantas. Assim, objetivou-se avaliar, neste trabalho, a resposta da cultura da soja proveniente de sementes tratadas com fungicidas na presença e ausência de polímeros em duas épocas de armazenamento e submetidas à inoculação com Bradyrhizobium em meio líquido e turfoso. Para tanto, as sementes foram tratadas com: carbendazin + thiram e thiabendazole + thiram via peliculização ou sem polímero, antes e após seis meses de armazenamento. Na semeadura as sementes foram submetidas ao tratamento com inoculante turfoso e líquido, ambos contendo estirpes de bactérias fixadoras deN2 atmosférico do gênero Bradyrhizobium (SEMIA 5079 e SEMIA 5019 ou foram semeadas sem inoculação. Utilizou-se o delineamento em blocos completos casualizados, com três repetições. Foram avaliados o número de nódulos, o teor de clorofila e o número de vagens. A inoculação das sementes com inoculante turfoso propiciou a formação de maior número de nódulos. Os tratamentos carbendazin + thiram e thiabendazole + thiram, independente da peliculização, reduziram o número de nódulos e carbendazin + thiram via peliculização diminuiu o número de vagem em plantas de soja. = Chemical treatment and seeds inoculation with Bradyrhizobium are used frequently in soybean, however the interaction between these practices can reduce nitrogen biological fixation, plants growth and development. The aim of thisstudy was to evaluate the response of soybean seeds treated with fungicides, with or without polymer, in two storage periods andinoculated with Bradyrhizobium using liquid or peat inoculant. Seeds were treated with: carbendazin + thiram or thiabendazole+ thiram by film

  12. Compatibilidade do tratamento de sementes de feijão-caupi com fungicidas e inoculação com estirpes de Bradyrhizobium Compatibility of the cowpea seed treatment with fungicides and inoculation with Bradyrhizobium strains

    Directory of Open Access Journals (Sweden)

    Manoel Luiz da Silva Neto

    2013-01-01

    Full Text Available O objetivo deste trabalho foi avaliar a compatibilidade do tratamento de sementes com fungicidas e a inoculação com estirpes de Bradyrhizobium em feijão-caupi. Em laboratório, avaliou-se a sobrevivência de células nas sementes da cultivar BRS Guariba, tratadas ou não com fungicidas (fludioxonil, carbendazim, carbendazim + thiram e carboxin + thiram e inoculadas ou não com Bradyrhizobium (estirpes UFLA3-84, BR 3267, INPA3-11B e BR 3262. Em casa de vegetação, conduziu-se experimento em vasos de Leonard, com os mesmos tratamentos. Foram avaliados: massa de matéria seca da parte aérea, além de número e massa de nódulos 25 dias após a emergência das plantas. No campo, dois experimentos foram conduzidos, tendo-se utilizado a estirpe BR 3262, com aplicação de fungicidas nas sementes: um em área de primeiro cultivo e outro em área cultivada anteriormente com culturas anuais. Avaliaram-se, aos 35 dias, o número de nódulos, a massa de nódulos secos e a massa de matéria seca da parte aérea, e, na colheita, a produtividade de grãos. Os fungicidas não tiveram efeito significativo sobre a sobrevivência de Bradyrhizobium, a nodulação das plantas e o rendimento de grãos, que, em média, foi superior a 1.200 kg ha-1. O tratamento de sementes de feijão-caupi com fungicidas é compatível com a inoculação das estirpes avaliadas.The objective of this work was to evaluate the compatibility of seed treatment with fungicides and Bradyrhizobium strains inoculation for cowpea. Cell survival on seeds of cultivar BRS Guariba treated or not with fungicides (fludioxonil, carbendazim, carbendazim + thiram, and carboxin + thiram and inoculated or not with Bradyrhizobium (strains UFLA3-84, BR 3267, INPA3-11B, and BR 3262 were evaluated in laboratory. Another experiment was carried out in greenhouse, in Leonard jars, using the same treatments. The following were evaluated: shoot dry matter weight, besides number and dry matter weight of

  13. Spatial risk profiling of Schistosoma japonicum in Eryuan county, Yunnan province, China

    Directory of Open Access Journals (Sweden)

    Peter Steinmann

    2007-11-01

    Full Text Available Bayesian spatial risk profiling holds promise to enhance our understanding of the epidemiology of parasitic diseases, and to target interventions in a cost-effective manner. Here, we present findings from a study using Bayesian variogram models to map and predict the seroprevalence of Schistosoma japonicum in Eryuan county, Yunnan province, China, including risk factor analysis. Questionnaire and serological data were obtained through a cross-sectional survey carried out in 35 randomly selected villages with 3,220 people enrolled. Remotely-sensed environmental data were derived from publicly available databases. Bivariate and non-spatial Bayesian multiple logistic regression models were used to identify associations between the local seroprevalence and demographic (i.e. age and sex, environmental (i.e. location of village, altitude, slope, land surface temperature and normalized difference vegetation index and socio-economic factors. In the spatially-explicit Bayesian model, S. japonicum seroprevalence was significantly associated with sex, age and the location of the village. Males, those aged below 10 years and inhabitants of villages situated on steep slopes (inclination ≥20° or on less precipitous slopes of >5° above 2,150 m were at lower risk of seroconversion than their respective counterparts. Our final prediction model revealed an elevated risk for seroconversion in the plains of the eastern parts of Eryuan county. In conclusion, the prediction map can be utilized for spatial targeting of schistosomiasis control interventions in Eryuan county. Moreover, S. japonicum seroprevalence studies might offer a convenient means to assess the infection pressure experienced by local communities, and to improve risk profiling in areas where the prevalence and infection intensities have come down following repeated rounds of praziquantel administration.

  14. Comparative Analysis of Proteome-Wide Lysine Acetylation in Juvenile and Adult Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    Qing Li

    2017-11-01

    Full Text Available Schistosomiasis is a devastating parasitic disease caused by tremotodes of the genus Schistosoma. Eggs produced by sexually mature schistosomes are the causative agents of for pathogenesis and transmission. Elucidating the molecular mechanism of schistosome development and sexual maturation would facilitate the prevention and control of schistosomiasis. Acetylation of lysine is a dynamic and reversible post-translational modification playing keys role in many biological processes including development in both eukaryotes and prokaryotes. To investigate the impacts of lysine acetylation on Schistosoma japonicum (S. japonicum development and sexual maturation, we used immunoaffinity-based acetyllysine peptide enrichment combined with mass spectrometry (MS, to perform the first comparative analysis of proteome-wide lysine acetylation in both female and male, juvenile (18 days post infection, 18 dpi and adult (28 dpi schistosome samples. In total, we identified 874 unique acetylated sites in 494 acetylated proteins. The four samples shared 47 acetylated sites and 46 proteins. More acetylated sites and proteins shared by both females and males were identified in 28 dpi adults (189 and 143, respectively than in 18 dpi schistosomula (76 and 59, respectively. More stage-unique acetylated sites and proteins were also identified in 28 dpi adults (494 and 210, respectively than in 18 dpi schistosomula (73 and 44, respectively. Functional annotation showed that in different developmental stages and genders, a number of proteins involving in muscle movement, glycometabolism, lipid metabolism, energy metabolism, environmental stress resistance, antioxidation, etc., displayed distinct acetylation profiles, which was in accordance with the changes of their biological functions during schistosome development, suggesting that lysine acetylation modification exerted important regulatory roles in schistosome development. Taken together, our data provided the first

  15. Identification and functional characterisation of a Schistosoma japonicum insulin-like peptide.

    Science.gov (United States)

    Du, Xiaofeng; McManus, Donald P; Cai, Pengfei; Hu, Wei; You, Hong

    2017-04-14

    Previous studies have shown that insulin receptors in schistosomes, triggered by host insulin, play an important role in parasite growth, development and fecundity by regulating glucose metabolism. However, limited information is available on the recently identified endogenous insulin-like peptide (ILP) in blood flukes. We isolated ILPs from Schistosoma japonicum (SjILP) and S. recognised (SmILP) and present results of their molecular and structural analysis. SjILP shares 63% amino acid identity with SmILP, but only 18% identity with human insulin. There is high cross immunological reactivity between the S. japonicum and S. mansoni ILPs as observed in western blots using an anti-SjILP polyclonal antibody. ADP binding/hydrolysis ability was observed in both SjILP and SmILP, but not in human insulin, suggesting a parasite-specific role for ILP compared with host insulin. Protein binding assays using the Octet-RED system showed SjILP binds S. japonicum IRs (SjIR1 and SjIR2) strongly. An anti-phospho antibody against extracellular signal-regulated kinase (Erk) recognised a 44-kDa target band in an extract of adult worms after stimulation by rSjILP in vitro, suggesting an important role for SjILP in activating SjIRs and in regulating downstream signal transduction. Immunolocalisation showed SjILP is located on the tegument and the underlying musculature, similar to that observed for SjIR1, but it is also present throughout the parenchyma of males and in the vitelline cells of females, the same locations as SjIR2 described in an earlier published study of ours. The same localisation of SjILP and the SjIRs is suggestive of an interaction between the insulin-like peptide and the IRs. In addition to binding host insulin, schistosomes also can express their own endogenous ILPs, which can activate the parasite insulin signal pathway, thereby playing a critical role in worm growth, development and fertility. These findings shed new light on ILPs in schistosomes, providing

  16. Identification of Host Insulin Binding Sites on Schistosoma japonicum Insulin Receptors.

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    Rachel J Stephenson

    Full Text Available Schistosoma japonicum insulin receptors (SjIRs have been identified as encouraging vaccine candidates. Interrupting or blocking the binding between host insulin and the schistosome insulin receptors (IRs may result in reduced glucose uptake leading to starvation and stunting of worms with a reduction in egg output. To further understand how schistosomes are able to exploit host insulin for development and growth, and whether these parasites and their mammalian hosts compete for the same insulin source, we identified insulin binding sites on the SjIRs. Based on sequence analysis and the predicted antigenic structure of the primary sequences of the SjIRs, we designed nine and eleven peptide analogues from SjIR-1 and SjIR-2, respectively. Using the Octet RED system, we identified analogues derived from SjIR-1 (10 and SjIR-2 (20, 21 and 22 with insulin-binding sequences specific for S. japonicum. Nevertheless, the human insulin receptor (HIR may compete with the SjIRs in binding human insulin in other positions which are important for HIR binding to insulin. However, no binding occurred between insulin and parasite analogues derived from SjIR-1 (2, 7 and 8 and SjIR-2 (14, 16 and 18 at the same locations as HIR sequences which have been shown to have strong insulin binding affinities. Importantly, we found two analogues (1 and 3, derived from SjIR-1, and two analogues (13 and 15 derived from SjIR-2, were responsible for the major insulin binding affinity in S. japonicum. These peptide analogues were shown to have more than 10 times (in KD value stronger binding capacity for human insulin compared with peptides derived from the HIR in the same sequence positions. Paradoxically, analogues 1, 3, 13 and 15 do not appear to contain major antigenic determinants which resulted in poor antibody responses to native S. japonicum protein. This argues against their future development as peptide-vaccine candidates.

  17. Ca. Nitrososphaera and Bradyrhizobium are inversely correlated and related to agricultural practices in long-term field experiments

    Directory of Open Access Journals (Sweden)

    Kateryna eZhalnina

    2013-05-01

    Full Text Available Agricultural land management, such as fertilization, liming, and tillage affects soil properties, including pH, organic matter content, nitrification rates, and the microbial community. Three different study sites were used to identify microorganisms that correlate with agricultural land use and to determine which factors regulate the relative abundance of the microbial signatures of the agricultural land-use. The three sites included in this study are the Broadbalk Experiment at Rothamsted Research, UK, the Everglades Agricultural Area, Florida, USA and the Kellogg Biological Station, Michigan, USA. The effects of agricultural management on the abundance and diversity of bacteria and archaea were determined using high throughput, barcoded 16S rRNA sequencing. In addition, the relative abundance of these organisms was correlated with soil features. Two groups of microorganisms involved in nitrogen cycle were highly correlated with land use at all three sites. The ammonia oxidizing-archaea, dominated by Ca. Nitrososphaera, were positively correlated with agriculture while a ubiquitous group of soil bacteria closely related to the diazotrophic symbiont, Bradyrhizobium, was negatively correlated with agricultural management. Analysis of successional plots showed that the abundance of ammonia oxidizing-archaea declined and the abundance of bradyrhizobia increased with time away from agriculture. This observation suggests that the effect of agriculture on the relative abundance of these genera is reversible. Soil pH and NH3 concentrations were positively correlated with archaeal abundance but negatively correlated with the abundance of Bradyrhizobium. The high correlations of Ca. Nitrososphaera and Bradyrhizobium abundances with agricultural management at three long-term experiments with different edaphoclimatic conditions allowed us to suggest these two genera as signature microorganisms for agricultural land use.

  18. Characteristics of granuloma formation and liver fibrosis in murine schistosomiasis mekongi: a morphological comparison between Schistosoma mekongi and S. japonicum infection.

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    Shimada, M; Kirinoki, M; Shimizu, K; Kato-Hayashi, N; Chigusa, Y; Kitikoon, V; Pongsasakulchoti, P; Matsuda, H

    2010-10-01

    A histopathological study was performed to clarify the characteristics of granuloma formation and liver fibrosis in Schistosoma mekongi infection in comparison with S. japonicum infection. Mice were exposed to S. mekongi (Laotian strain) and S. japonicum (Japanese strain) cercariae, and were dissected at 6, 8, 12, 16, and 20 weeks post-exposure. In the liver, granulomas in S. mekongi infection were cellular, initially organized with foam cells, and continuously appeared in the intralobular area, while granulomas in S. japonicum infection were fibrous and did not continuously appear in the intralobular area. Portal fibrosis was not seen in S. mekongi infection, but was commonly seen in S. japonicum infection in the later weeks. Granulomas in the small intestine were seen mainly in the submucosa with foam cells in S. mekongi infection and without foam cells in S. japonicum infection. The lung granulomas contained mainly histiocytes in both S. mekongi and S. japonicum infection. The absence of portal fibrosis in S. mekongi infection allows schistosome eggs to infiltrate into the intralobular area continuously, which can be what lies behind the ultrasonographic differences; the echogenic network pattern as was seen in S. japonicum infection, has not been noted in S. mekongi infection.

  19. Draft genome sequence of Bradyrhizobium paxllaeri LMTR 21T isolated from Lima bean (Phaseolus lunatus in Peru

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    Ernesto Ormeño-Orrillo

    2017-09-01

    Full Text Available Bradyrhizobium paxllaeri is a prevalent species in root nodules of the Lima bean (Phaseolus lunatus in Peru. LMTR 21T is the type strain of the species and was isolated from a root nodule collected in an agricultural field in the Peruvian central coast. Its 8.29 Mbp genome encoded 7635 CDS, 71 tRNAs and 3 rRNAs genes. All genes required to stablish a nitrogen-fixing symbiosis with its host were present. The draft genome sequence and annotation have been deposited at GenBank under the accession number MAXB00000000.

  20. Induction of specific immunity against Schistosoma japonicum by exposure of mice to ultraviolet attenuated cercariae

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    Moloney, N.A.; Bickle, Q.D.; Webbe, G. (London School of Hygiene and Tropical Medicine, St. Albans (UK). Winches Farm Field Station)

    1985-01-01

    Mice can be partially protected against Schistosoma japonicum by prior exposure to ultraviolet (UV)-attenuated infections which fail to survive to the adult stage and produce no overt pathology in the host. Optimum resistance was induced by parasites exposed to 40 seconds of UV, significantly lower levels of resistance being stimulated by both shorter and longer exposures. No consistent relationship between the degree of resistance induced and the number of irradiated cercariae given could be demonstrated and equivocal results were obtained when comparing the efficacy of single and multiple vaccinations. Vaccinations with UV-attenuated cercariae given intraperitoneally (i.p.) were as efficacious as those given percutaneously but mice were as or more resistant to challenges given by the i.p. route, the possible reasons are discussed. There was no observed delay in the migration of the challenge, vaccinated mice being as resistant when perfused 6 or 3.5 weeks after challenge. Vaccination was species specific since mice exposed to either UV-attenuated S. japonicum cercariae or gamma-attenuated S. mansoni cercariae were resistant to homologous but not heterologous challenge.

  1. Occurrence of Rust on Peucedanum japonicum Caused by Puccinia jogashimensis in Korea

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    Sug-Ju Ko

    2015-12-01

    Full Text Available During July to November 2014, severe rust infection was consistently found on Peucedanum japonicum growing farm in Yeosu, Korea. The rust was observed mainly on lower leaf surfaces. Symptoms of typical plants included yellow-orange rust pustules were observed on the petiole and leaf surface with small yellowish to chlorotic lesions on the upper surface. No symptom was observed on flowers. Uredinia were occurred amphigenous on leaf surface, and occasionally caulicolous, scattered or loosely aggregate, rounded to oblong, 0.4 to 4 mm in diameter, covered by epidermis, then naked, surrounded by ruptured epidermis, pulverulent, and brown. Urediniospores were ovate-ellipsoid, ellipsoid or subglobose, light brown, 20 to 45 ×15 to 35 µm, walls 2 to 4 µm thick. The resulting sequences were deposited in GenBank with accession No. KT778808, KT778809, and KT778810, respectively. Since this was the first accession of 28S sequence Puccinia jogashimensis, there was no exact match in GenBank nucleotide database. On the basis of the morphological characteristics and phylogenetic analyses of 28S rDNA, the fungus was identified as P. jogashimensis. To our knowledge, this is the first confirmed report on the occurrence of P. jogashimensis on P. japonicum in Korea.

  2. Intra- and interspecies transfer and expression of Rhizobium japonicum hydrogen uptake genes and autotrophic growth capability.

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    Lambert, G R; Cantrell, M A; Hanus, F J; Russell, S A; Haddad, K R; Evans, H J

    1985-05-01

    Cosmids containing hydrogen uptake genes have previously been isolated in this laboratory. Four new cosmids that contain additional hup gene(s) have now been identified by conjugal transfer of a Rhizobium japonicum 122DES gene bank into a Tn5-generated Hup(-) mutant and screening for the acquisition of Hup activity. The newly isolated cosmids, pHU50-pHU53, contain part of the previously isolated pHU1 but extend as far as 20 kilobases beyond its border. pHU52 complements five of six Hup(-) mutants and confers activity on several Hup(-) wild-type R. japonicum strains in the free-living state and where tested in nodules. Transconjugants obtained from interspecies transfer of pHU52 to Rhizobium meliloti 102F28, 102F32, and 102F51 and Rhizobium leguminosarum 128C53 showed hydrogen-dependent methyleneblue reduction, performed the oxyhydrogen reaction, and showed hydrogen-dependent autotrophic growth by virtue of the introduced genes. The identity of the presumptive transconjugants was confirmed by antibiotic-resistance profiles and by plant nodulation tests.

  3. Spatial distribution of human Schistosoma japonicum infections in the Dongting Lake Region, China.

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    Giovanna Raso

    Full Text Available BACKGROUND: The aim of this study was to spatially model the effect of demographic, reservoir hosts and environmental factors on human Schistosoma japonicum infection prevalence in the Dongting Lake area of Hunan Province, China and to determine the potential of each indicator in targeting schistosomiasis control. METHODOLOGY/PRINCIPAL FINDINGS: Cross-sectional serological, coprological and demographic data were obtained from the 2004 nationwide periodic epidemiologic survey for Hunan Province. Environmental data were downloaded from the USGS EROS data centre. Bayesian geostatistical models were employed for spatial analysis of the infection prevalence among study participants. A total of 47,139 participants from 47 administrative villages were selected. Age, sex and occupation of residents and the presence of infected buffaloes and environmental factors, i.e. NDVI, distance to the lake and endemic type of setting, were significantly associated with S. japonicum infection prevalence. After taking into account spatial correlation, however, only demographic factors (age, sex and occupation and the presence of infected buffaloes remained significant indicators. CONCLUSIONS/SIGNIFICANCE: Long established demographic factors, as well presence of host reservoirs rather than environmental factors are driving human transmission. Findings of this work can be used for epidemiologic surveillance and for the future planning of interventions in the Dongting Lake area of Hunan Province.

  4. Identification and characterization of microRNAs and endogenous siRNAs in Schistosoma japonicum

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    Wang Heng

    2010-01-01

    Full Text Available Abstract Background Small endogenous non-coding RNAs (sncRNAs such as small interfering RNA (siRNA, microRNA and other small RNA transcripts are derived from distinct loci in the genome and play critical roles in RNA-mediated gene silencing mechanisms in plants and metazoa. They are approximately 22 nucleotides long; regulate mRNA stability through perfect or imperfect match to the targets. The biological activities of sncRNAs have been related to many biological events, from resistance to microbe infections to cellular differentiation. The development of the zoonotic parasite Schistosoma japonicum parasite includes multiple steps of morphological alterations and biological differentiations, which provide a unique model for studies on the functions of small RNAs. Characterization of the genome-wide transcription of the sncRNAs will be a major step in understanding of the parasite biology. The objective of this study is to investigate the transcriptional profile and potential function of the small non-coding RNAs in the development of S. japanicum. Results The endogenous siRNAs were found mainly derived from transposable elements (TE or transposons and the natural antisense transcripts (NAT. In contrast to other organisms, the TE-derived siRNAs in S. japonicum were more predominant than other sncRNAs including microRNAs (miRNAs. Further, there were distinct length and 3'end variations in the sncRNAs, which were associated with the developmental differentiation of the parasite. Among the identified miRNA transcripts, there were 38 unique to S. japonicum and 16 that belonged to 13 miRNA families are common to other metazoan lineages. These miRNAs were either ubiquitously expressed, or they exhibited specific expression patterns related to the developmental stages or sex. Genes that encoded miRNAs are mainly located in clusters within the genome of S. japonicum. However, genes within one cluster could be differentially transcribed, which suggested

  5. Bayesian spatio-temporal modeling of Schistosoma japonicum prevalence data in the absence of a diagnostic 'gold' standard.

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    Xian-Hong Wang

    Full Text Available BACKGROUND: Spatial modeling is increasingly utilized to elucidate relationships between demographic, environmental, and socioeconomic factors, and infectious disease prevalence data. However, there is a paucity of studies focusing on spatio-temporal modeling that take into account the uncertainty of diagnostic techniques. METHODOLOGY/PRINCIPAL FINDINGS: We obtained Schistosoma japonicum prevalence data, based on a standardized indirect hemagglutination assay (IHA, from annual reports from 114 schistosome-endemic villages in Dangtu County, southeastern part of the People's Republic of China, for the period 1995 to 2004. Environmental data were extracted from satellite images. Socioeconomic data were available from village registries. We used Bayesian spatio-temporal models, accounting for the sensitivity and specificity of the IHA test via an equation derived from the law of total probability, to relate the observed with the 'true' prevalence. The risk of S. japonicum was positively associated with the mean land surface temperature, and negatively correlated with the mean normalized difference vegetation index and distance to the nearest water body. There was no significant association between S. japonicum and socioeconomic status of the villages surveyed. The spatial correlation structures of the observed S. japonicum seroprevalence and the estimated infection prevalence differed from one year to another. Variance estimates based on a model adjusted for the diagnostic error were larger than unadjusted models. The generated prediction map for 2005 showed that most of the former and current infections occur in close proximity to the Yangtze River. CONCLUSION/SIGNIFICANCE: Bayesian spatial-temporal modeling incorporating diagnostic uncertainty is a suitable approach for risk mapping S. japonicum prevalence data. The Yangtze River and its tributaries govern schistosomiasis transmission in Dangtu County, but spatial correlation needs to be taken

  6. Bayesian spatio-temporal modeling of Schistosoma japonicum prevalence data in the absence of a diagnostic 'gold' standard.

    Science.gov (United States)

    Wang, Xian-Hong; Zhou, Xiao-Nong; Vounatsou, Penelope; Chen, Zhao; Utzinger, Jürg; Yang, Kun; Steinmann, Peter; Wu, Xiao-Hua

    2008-06-11

    Spatial modeling is increasingly utilized to elucidate relationships between demographic, environmental, and socioeconomic factors, and infectious disease prevalence data. However, there is a paucity of studies focusing on spatio-temporal modeling that take into account the uncertainty of diagnostic techniques. We obtained Schistosoma japonicum prevalence data, based on a standardized indirect hemagglutination assay (IHA), from annual reports from 114 schistosome-endemic villages in Dangtu County, southeastern part of the People's Republic of China, for the period 1995 to 2004. Environmental data were extracted from satellite images. Socioeconomic data were available from village registries. We used Bayesian spatio-temporal models, accounting for the sensitivity and specificity of the IHA test via an equation derived from the law of total probability, to relate the observed with the 'true' prevalence. The risk of S. japonicum was positively associated with the mean land surface temperature, and negatively correlated with the mean normalized difference vegetation index and distance to the nearest water body. There was no significant association between S. japonicum and socioeconomic status of the villages surveyed. The spatial correlation structures of the observed S. japonicum seroprevalence and the estimated infection prevalence differed from one year to another. Variance estimates based on a model adjusted for the diagnostic error were larger than unadjusted models. The generated prediction map for 2005 showed that most of the former and current infections occur in close proximity to the Yangtze River. Bayesian spatial-temporal modeling incorporating diagnostic uncertainty is a suitable approach for risk mapping S. japonicum prevalence data. The Yangtze River and its tributaries govern schistosomiasis transmission in Dangtu County, but spatial correlation needs to be taken into consideration when making risk prediction at small scales.

  7. Effects of Cu on the content of chlorophylls and secondary metabolites in the Cu-hyperaccumulator lichen Stereocaulon japonicum.

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    Nakajima, Hiromitsu; Hara, Kojiro; Yamamoto, Yoshikazu; Itoh, Kiminori

    2015-03-01

    Understanding the relationship between Cu and Cu-hyperaccumulator lichens is important for their application in monitoring and assessing heavy metal pollution. We investigated the Cu-hyperaccumulator lichen Stereocaulon japonicum at several Cu-polluted and control sites in Japan, and found the lichen to be widely distributed. Its concentrations of Cu, chlorophylls, and secondary metabolites, chlorophyll-related indices, and absorption spectra were measured, and we observed negative effects of Cu on these concentrations and indices. For highly Cu-polluted samples (>100ppm dry weight), however, we found significant linear correlations between Cu and chlorophyll concentrations. This can be considered as the response of the photobiont in S. japonicum to Cu stress. In highly Cu-polluted samples the chlorophyll-related indices and concentration of total secondary metabolites were almost constant regardless of Cu concentration. This suggests that the increase in chlorophyll concentration with the increase in Cu concentration enhances photosynthetic productivity per unit biomass, which will allow the production of extra structure and energy for maintaining the chlorophyll-related indices under Cu stress. The relationship between the increase in chlorophyll concentration of S. japonicum and the decrease in secondary metabolite concentration of the lichen can be explained by considering the balance of carbohydrates in the lichen. We found that a spectral index A372-A394 can be a useful index of the concentrations of Cu and total secondary metabolites in S. japonicum. These findings show the adjustment of the content of chlorophylls and secondary metabolites in S. japonicum to Cu stress, and provide a better understanding of the relationship between Cu and the Cu-hyperaccumulator lichen. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Peanut plant growth and yield as influenced by co-inoculation with Bradyrhizobium and some rhizo-microorganisms under sandy loam soil conditions

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    F.Sh.F. Badawi

    2011-06-01

    Full Text Available The ability of tested rhizomicrobial isolates (Serratia marcescens and Trichoderma harzianum along with a strain of root nodule bacteria (Bradyrhizobium spp. to exhibit some PGP-properties was evaluated in vitro conditions. The main PGP-properties, namely the ability to solubilize-P and production of IAA, as well as production of siderophores and HCN were examined. Additionally, field trials were conducted on sandy loam soil at El-Tahrir Province during two successive summer seasons to study the effect of co-inoculation with Bradyrhizobium either individually or together with S. marcescens and/or T. harzianum on nodulation, some plant growth characters, peanut yield and its yield components. The in vitro experiment revealed that all of the tested microorganisms were apparently able to trigger PGP-properties. Phosphate solubilization was the common feature of the employed microorganisms. However, T. harzianum appeared to be superior to other microorganisms, and Bradyrhizobium displayed the lowest capacity. The ability of the microorganisms to produce indole compounds showed that S. marcescens was more effective in IAA production and followed by Bradyrhizobium. Capacity of S. marcescens and T. harzianum to excrete ferric-specific ligands (siderophores and HCN was detected, while Bradyrhizobium failed to produce such compounds. Results of field trials showed that the uninoculated peanut had the least nodulation status, N2-ase activity and all vegetative growth characters in both studied seasons. Bacterization of peanut seeds with bradyrhizobia exerted considerable improvement in number and mass of root nodules, increased the rate of acetylene reduction and all growth characters in comparison to the uninoculated control. The synergy inoculation between bradyrhizobia and any of the tested microorganisms led to further increases of all mentioned characters and strengthened the stimulating effect of the bacterial inoculation. However, the promotive

  9. Immunization of pigs against infection with Schistosoma japonicum using ultraviolet-attenuated cercariae

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    Shi, Y.-E.; Jiang, C.-F.; Han, J.-J.; Li, Y.-L. (Tongji Medical Univ., Wuhan (China). Dept. of Parasitology); Ruppel, A. (Institute for Tropical Hygiene, Heidelberg (Germany))

    1993-06-01

    Since pigs are important in the zoonotic transmission of schistosomiasis japonica in China, a veterinary vaccine might contribute to the control of the disease in humans. Pigs were immunized with three doses each of 10 000 cercariae of Schistosoma japonicum attenuated with ultraviolet light (400 [mu]Watt.min/cm[sup 2]). The experiment was performed with portable irradiation equipment in a rural area of the Hubei Province (P.R. China). A challenge infection of 1000 untreated cercariae was given 2.5 or 6 months after the last immunization, and age-matched naive pigs were challenged as a control. Immunized pigs developed about 90% resistance against the challenge. The liver egg load of these animals was reduced by over 90%. Less than 0.01% of the immunizing cercariae developed to adult parasites and the vaccination had no apparent adverse influence on the pig's health. (Author).

  10. Pararosaniline pamoate (CI-403-A) in the treatment of Schistosoma japonicum infection in the Philippines.

    Science.gov (United States)

    Pesigan, T P; Banzon, T C; Santos, A T; Noseñas, J; Zabala, R G

    1967-01-01

    Trials have been carried out, first on a relatively small scale among patients in Manila and later on a larger scale among domiciliary patients in an area of endemic schistosomiasis in Leyte Province, Philippines, with various dosage schedules of pararosaniline pamoate (CI-403-A) to determine that drug's efficacy and optimum dosage against Schistosoma japonicum infection.Given orally in gelatin capsules, the drug was well tolerated even in children, with few side-effects, and was both curative and suppressive when administered in a maximum dosage of 35-40 mg/kg body-weight per day for as many as 52 days spread over a total treatment period of 203 days.The authors recommend its use for mass treatment, especially among schoolchildren, in combination with other established schistosomiasis control measures-health education, environmental sanitation, and snail control.

  11. Oxyleghemoglobin-mediated Hydrogen Oxidation by Rhizobium japonicum USDA 122 DES Bacteroids.

    Science.gov (United States)

    Emerich, D W; Albrecht, S L; Russell, S A; Ching, T; Evans, H J

    1980-04-01

    Oxyleghemoglobin was used to supply low concentrations of O(2) to H(2)-oxidizing bacteroids from Rhizobium japonicum USDA 122 DES. The H(2) oxidation system of these bacteroids was capable of effectively utilizing O(2) at the low concentrations of O(2) expected to be found in soybean nodules. Apparent K(m) values of approximately 10 nanomolar O(2) have been calculated for the oxyhydrogen reaction. These values include the K(m) values for both H(2) oxidation and endogenous substrate oxidation. Even in the presence of oxyleghemoglobin, H(2) additions stimulated C(2)H(2) reduction, reduced the rate of endogenous respiration and maintained the ATP contents of bacteroids. In our reconstituted oxyleghemoglobin and bacteriod system, we estimate that the H(2) oxidation system is capable of recycling all of the H(2) evolved during the N(2) fixation process.

  12. Oxyleghemoglobin-mediated Hydrogen Oxidation by Rhizobium japonicum USDA 122 DES Bacteroids 1

    Science.gov (United States)

    Emerich, David W.; Albrecht, Steve L.; Russell, Sterling A.; Ching, Temay; Evans, Harold J.

    1980-01-01

    Oxyleghemoglobin was used to supply low concentrations of O2 to H2-oxidizing bacteroids from Rhizobium japonicum USDA 122 DES. The H2 oxidation system of these bacteroids was capable of effectively utilizing O2 at the low concentrations of O2 expected to be found in soybean nodules. Apparent Km values of approximately 10 nanomolar O2 have been calculated for the oxyhydrogen reaction. These values include the Km values for both H2 oxidation and endogenous substrate oxidation. Even in the presence of oxyleghemoglobin, H2 additions stimulated C2H2 reduction, reduced the rate of endogenous respiration and maintained the ATP contents of bacteroids. In our reconstituted oxyleghemoglobin and bacteriod system, we estimate that the H2 oxidation system is capable of recycling all of the H2 evolved during the N2 fixation process. PMID:16661247

  13. Quinone oxidoreductase 2 is involved in haustorium development of the parasitic plant Phtheirospermum japonicum.

    Science.gov (United States)

    Ishida, Juliane K; Yoshida, Satoko; Shirasu, Ken

    2017-07-03

    The family Orobanchaceae includes many parasitic plant species. Parasitic plants invade host vascular tissues and form organs called haustoria, which are used to obtain water and nutrients. Haustorium formation is initiated by host-derived chemicals including quinones and flavonoids. Two types of quinone oxidoreductase (QR) are involved in signal transduction leading to haustorium formation; QR1 mediates single-electron transfers and QR2 mediates 2-electron transfers. In the facultative parasite Triphysaria versicolor, QR1 is involved in haustorium induction signaling, while this role is played by QR2 in the model plant Phtheirospermum japonicum. Our results suggest that there is functional diversification in haustorium signaling molecules among different species of the Orobanchaceae.

  14. [Activities of treg cells stimulated by soluble adult worm antigen and egg antigen of Schistosoma japonicum].

    Science.gov (United States)

    Dong, Xiao-Xiao; Zhang, Cui; Yang, Xiao-Wei; Li, Yong; Chen, Xiao-Jun; Xue, Xue; Zhang, Wei-Wei; Xu, Zhi-Peng; Kong, Wen-Jun; Zhu, Ji-Feng; Zhou, Sha; Liu, Feng; Su, Chuan

    2013-04-01

    To observe and compare the effects of soluble adult worm antigen (SWA) and soluble egg antigen (SEA) of Schistosoma japonicum on the induction of Treg cells and the suppressive activity of Treg cells. Splenocytes were prepared from mice treated with PBS, SWA, and SEA, respectively, and then the proportions of Treg cells and the levels of IL-10 and TGF-beta in Treg cells were determined by FACS. The purified Treg cells from the mice treated as above-mentioned were detected for their immunosuppressive activities by incorporation of [3H] thymidine for the final 16 h of culture. Compared to SWA, SEA induced the higher proportion of Treg cells with a stronger suppressive activity, which produced the higher levels of IL-10 and TGF-beta (P < 0.05). SEA significantly induces Treg cells and enhances their immunosuppressive activity.

  15. Treatment for Schistosoma japonicum, reduction of intestinal parasite load, and cognitive test score improvements in school-aged children.

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    Ezeamama, Amara E; McGarvey, Stephen T; Hogan, Joseph; Lapane, Kate L; Bellinger, David C; Acosta, Luz P; Leenstra, Tjalling; Olveda, Remigio M; Kurtis, Jonathan D; Friedman, Jennifer F

    2012-01-01

    To determine whether treatment of intestinal parasitic infections improves cognitive function in school-aged children, we examined changes in cognitive testscores over 18 months in relation to: (i) treatment-related Schistosoma japonicum intensity decline, (ii) spontaneous reduction of single soil-transmitted helminth (STH) species, and (iii) ≥2 STH infections among 253 S. japonicum-infected children. Helminth infections were assessed at baseline and quarterly by the Kato-Katz method. S. japonicum infection was treated at baseline using praziquantel. An intensity-based indicator of lower vs. no change/higher infection was defined separately for each helminth species and joint intensity declines of ≥2 STH species. In addition, S. japonicum infection-free duration was defined in four categories based on time of schistosome re-infection: >18 (i.e. cured), >12 to ≤18, 6 to ≤12 and ≤6 (persistently infected) months. There was no baseline treatment for STHs but their intensity varied possibly due to spontaneous infection clearance/acquisition. Four cognitive tests were administered at baseline, 6, 12, and 18 months following S. japonicum treatment: learning and memory domains of Wide Range Assessment of Memory and Learning (WRAML), verbal fluency (VF), and Philippine nonverbal intelligence test (PNIT). Linear regression models were used to relate changes in respective infections to test performance with adjustment for sociodemographic confounders and coincident helminth infections. Children cured (β = 5.8; P = 0.02) and those schistosome-free for >12 months (β = 1.5; P = 0.03) scored higher in WRAML memory and VF tests compared to persistently infected children independent of STH infections. A decline vs. no change/increase of any individual STH species (β:11.5-14.5; all Ptest independent of schistosome infection. Hookworm and Trichuris trichiura declines were independently associated with improvements in WRAML memory scores as was the joint

  16. Intake of Erythrocytes Required for Reproductive Development of Female Schistosoma japonicum

    Science.gov (United States)

    Wang, Jipeng; Wang, Shuqi; Liu, Xiufeng; Xu, Bin; Chai, Riyi; Zhou, Pan; Ju, Chuan; Sun, Jun; Brindley, Paul J.; Hu, Wei

    2015-01-01

    The reproductive development and maturation of female schistosomes are crucial since their released eggs are responsible for the host immunopathology and transmission of schistosomiasis. However, little is known about the nutrients required by female Schistosoma japonicum during its sexual maturation. We evaluated the promoting effect of several nutrients (calf serum, red blood cells (RBCs), ATP and hypoxanthine) on the reproductive development of pre-adult females at 18 days post infection (dpi) from mixed infections and at 50 dpi from unisexual infections of laboratory mice in basic medium RPMI-1640. We found RBCs, rather than other nutrients, promoted the female sexual maturation and egg production with significant morphological changes. In 27% of females (18 dpi) from mixed infections that paired with males in vitro on day 14, vitelline glands could be positively stained by Fast Blue B; and in 35% of females (50 dpi) from unisexual infections on day 21, mature vitelline cells were observed. Infertile eggs were detected among both groups. To analyze which component of mouse RBCs possesses the stimulating effect, RBCs were fractionated and included in media. However, the RBC fractions failed to stimulate development of the female reproductive organs. In addition, bovine hemoglobin hydrolysate, digested by neutral protease, was found to exhibit the promoting activity instead of untreated bovine hemoglobin. The other protein hydrolysate, lactalbumin hydrolysate, exhibited a similar effect with bovine hemoglobin hydrolysate. Using quantitative RT-PCR, we found the expression levels of four reproduction-related genes were significantly stimulated by RBCs. These data indicate that RBCs provide essential nutrients for the sexual maturation of female S. japonicum and that the protein component of RBCs appeared to constitute the key nutrient. These findings would improve laboratory culture of pre-adult schistosomes to adult worms in medium with well-defined components

  17. Intake of Erythrocytes Required for Reproductive Development of Female Schistosoma japonicum.

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    Jipeng Wang

    Full Text Available The reproductive development and maturation of female schistosomes are crucial since their released eggs are responsible for the host immunopathology and transmission of schistosomiasis. However, little is known about the nutrients required by female Schistosoma japonicum during its sexual maturation. We evaluated the promoting effect of several nutrients (calf serum, red blood cells (RBCs, ATP and hypoxanthine on the reproductive development of pre-adult females at 18 days post infection (dpi from mixed infections and at 50 dpi from unisexual infections of laboratory mice in basic medium RPMI-1640. We found RBCs, rather than other nutrients, promoted the female sexual maturation and egg production with significant morphological changes. In 27% of females (18 dpi from mixed infections that paired with males in vitro on day 14, vitelline glands could be positively stained by Fast Blue B; and in 35% of females (50 dpi from unisexual infections on day 21, mature vitelline cells were observed. Infertile eggs were detected among both groups. To analyze which component of mouse RBCs possesses the stimulating effect, RBCs were fractionated and included in media. However, the RBC fractions failed to stimulate development of the female reproductive organs. In addition, bovine hemoglobin hydrolysate, digested by neutral protease, was found to exhibit the promoting activity instead of untreated bovine hemoglobin. The other protein hydrolysate, lactalbumin hydrolysate, exhibited a similar effect with bovine hemoglobin hydrolysate. Using quantitative RT-PCR, we found the expression levels of four reproduction-related genes were significantly stimulated by RBCs. These data indicate that RBCs provide essential nutrients for the sexual maturation of female S. japonicum and that the protein component of RBCs appeared to constitute the key nutrient. These findings would improve laboratory culture of pre-adult schistosomes to adult worms in medium with well

  18. Bradyrhizobium spp. Strains in Symbiosis with Pigeon Pea cv. Fava-Larga under Greenhouse and Field Conditions

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    Márcia Rufini

    Full Text Available ABSTRACT: Optimization of symbiosis between nitrogen-fixing bacteria and legumes has been extensively studied, seeking agricultural sustainability. To evaluate the symbiotic efficiency of nitrogen-fixing bacterial strains belonging to the Bradyrhizobium genus with pigeon pea (Cajanus cajan (L. Millsp. cv. Fava-Larga, experiments were conducted in Leonard jars (axenic conditions, pots with soil, and in the field. Ten strains were tested in Leonard jars, and three strains, in addition to BR 29, were selected according to their ability to promote the growth of pigeon pea, for further tests in pots with different soil types (Inceptsol and Oxisol and in the field (Oxisol. Treatments were compared with strains BR 2003 and BR 2801 (approved as inoculants for pigeon pea, with a non-inoculated control with mineral N fertilization, and with another non-inoculated control (absolute control with low mineral N concentration (Leonard jars or without mineral N fertilization (soil. The efficiency of Bradyrhizobium strains in axenic conditions varies among strains, being higher when pigeon pea cv. Fava-Larga establishes symbiosis with the strains UFLA 03-320, UFLA 03-321, UFLA 04-212, BR 2801, and BR 2003. The soil type influences the symbiotic efficiency of Bradyrhizobium-pigeon pea in soil in the greenhouse, mainly in Inceptsol, in which strains UFLA 04-212, BR 2801, and BR 2003 increased N accumulation in the plant. The strain UFLA 03-320 increased shoot dry matter and N accumulation in the shoot equivalent to the mineral N treatment under field conditions. UFLA 03-320, BR 29, UFLA 03-321, and UFLA 04-212 promoted yields similar to those of the reference strain (BR 2801, and of the mineral N treatment with 70 kg ha-1 urea-N. These results confirm that pigeon pea establishes efficient symbiosis, which provides the N required for its growth. All strains, except for BR 2003, show potential for recommendation as inoculants for grain production. The strain UFLA 03

  19. Comportamiento de la simbiosis leguminosa-rizobio en Centrosema plumieri inoculada con Bradyrhizobium sp. Ensayo de campo Performance of the legume-rhizobium symbiosis in Centrosema plumieri inoculated with Bradyrhizobium sp. Field essay

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    C. J. Bécquer

    2012-06-01

    Full Text Available Se efectuó un experimento de campo, sin riego, en la Estación Experimental de Pastos y Forrajes de Sancti Spíritus, Cuba, durante el periodo de noviembre de 2001 a abril de 2002. El objetivo fue evaluar el comportamiento simbiótico de Centrosema plumieri, inoculada con cepas de Bradyrhizobium sp., previamente aisladas de leguminosas forrajeras procedentes de ecosistemas ganaderos de Cuba. El diseño experimental fue de bloques al azar; se utilizaron 11 tratamientos inoculados con cepas nativas, un testigo inoculado con la cepa de referencia 5030 (Bradyrhizobium sp. y un control no inoculado, con tres réplicas. Se aplicó un análisis de varianza (ANOVA, así como de correlación y regresión. Los valores de los tratamientos inoculados fueron estadísticamente iguales a los del control absoluto, en cuanto al peso seco aéreo (PSA y el rendimiento de N (RN, excepto el inoculado con la cepa HA2 en RN. En general, se demostró una débil efectividad de la inoculación. No obstante, en el índice de nodulación (IN sobresalieron JH2 y HA2, con valores estadísticamente superiores (pA field trial, without irrigation, was conducted at the Sancti Spiritus Experimental Station of Pastures and Forages, Cuba, from November, 2001, to April, 2002. The objective was to evaluate the symbiotic performance of Centrosema plumieri, inoculated with Bradyrhizobium sp. strains, previously isolated from forage legumes of Cuban livestock production ecosystems. The experimental design consisted in randomized blocks; 11 treatments inoculated with native strains, a control inoculated with the reference strain 5030 (Bradyrhizobium sp. and a non-inoculated control were used, with three replications. A variance analysis (ANOVA, as well as correlation and regression analyses, were applied. The values of the inoculated treatments were statistically equal to those of the absolute control, regarding aerial dry weight (ADW and N yield (NY, except the one inoculated with the

  20. [Preliminary study on establishing an animal model of neuroschistosomiasis by direct injection of Schistosoma japonicum eggs through skull].

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    Xu, Jia; Lu, Xiao-Jie; Wang, Dan; Wu, Ming-Can; Chen, Shi-Jie; Li, Jun-Chuan; Wang, Peng

    2013-02-01

    To establish an experimental model of neuroschistosomiasis and investigate the model establishment factors. Rabbits were used for the animal model and Schistosoma japonicum eggs (1 mg/ml) were directly injected into the brain by two ways of a bone drill or needle. The symptoms were observed and in the first and second week and later, the rabbits' brains were removed for pathological examinations. One to two weeks after the injection of schistosome eggs, the rabbits had various neurological symptoms such as loss of appetite, hemiparesis, seizure, etc. The pathological analysis showed the schistosome egg granuloma inflammatory reaction among 90% rabbits. This new method of direct injection of S. japonicum eggs through skull into the brain provides a good and easy animal model of neuroschistosomiasis.

  1. A comparative study of the vitelline cell in Schistosoma mansoni, S. haematobium, S. japonicum and S. mattheei.

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    Erasmus, D A; Popiel, I; Shaw, J R

    1982-04-01

    A comparison is given of the ultrastructure of the vitelline cell in Schistosoma mansoni, S. haematobium, S. japonicum and S. mattheei. Four stages in development of the vitelline cell have been categorized as follows: Stage 1, the undifferentiated cell; Stage 2, the developing cell showing the beginning of synthetic activity; Stage 3, the developing cell showing active protein synthesis; Stage 4, the fully mature vitelline cell. These stages in development have been defined morphologically and Stages 1, 2 and 3 are very similar in all 4 species. Lipid is present in the Stage 4 cells of all species but appears earlier at Stage 3 in S. haematobium and S. mattheei. There are several differences as to the intercellular inclusions of the Stage 4 cells, the most marked of these being the absence of calcareous corpuscles from S. japonicum as compared with the other 3 species.

  2. Comparison of worm development and host immune responses in natural hosts of schistosoma japonicum, yellow cattle and water buffalo

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    Yang Jianmei

    2012-03-01

    Full Text Available Abstract Background Yellow cattle and water buffalo are two of the most important natural hosts for Schistosoma japonicum in China. Previous observation has revealed that yellow cattle are more suited to the development of S. japonicum than water buffalo. Understanding more about the molecular mechanisms involved in worm development, as well as the pathological and immunological differences between yellow cattle and water buffalo post infection with S japonicum will provide useful information for the vaccine design and its delivery procedure. Results The worm length (p p p + T cells was higher in yellow cattle, while the percentage of CD8+ T cells was higher in water buffalo from pre-infection to 7 w post infection. The CD4/CD8 ratios were decreased in both species after challenge with schistosomes. Comparing with water buffalo, the IFN-γ level was higher and decreased significantly, while the IL-4 level was lower and increased gradually in yellow cattle from pre-infection to 7 w post infection. Conclusions In this study, we confirmed that yellow cattle were more suited to the development of S. japonicum than water buffalo, and more serious pathological damage was observed in infected yellow cattle. Immunological analysis suggested that CD4+ T cells might be an integral component of the immune response and might associate with worm development in yellow cattle. A shift from Th1 to Th2 type polarized immunity was only shown clearly in schistosome-infected yellow cattle, but no shift in water buffalo. The results provide valuable information for increased understanding of host-schistosome interactions, and for control of schistosomiasis.

  3. Inconsistent protective efficacy and marked polymorphism limits the value of Schistosoma japonicum tetraspanin-2 as a vaccine target.

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    Wenbao Zhang

    Full Text Available BACKGROUND: Schistosoma mansoni tetraspanin 2 (Sm-TSP-2 has been shown to be strongly recognized by IgG1 and IgG3 antibodies from individuals putatively resistant to schistosome infection, but not chronically infected people, and to induce high levels of protection against challenge infection in the murine model of schistosomiasis. Amplification by PCR of homologous sequences from male and female S. japonicum worms showed the presence of 7 different clusters or subclasses of S. japonicum TSP-2. We determined the protective efficacy of one subclass - Sj-TSP-2e. METHODOLOGY/PRINCIPAL FINDINGS: Following the alignment of 211 cDNAs, we identified 7 clusters encoding S. japonicum TSP-2 (Sj-TSP-2 based on sequence variation in the large extracellular loop (LEL region with differing frequency of transcription in male and female worms. Quantitative PCR analysis revealed elevated expression of Sj-TSP-2 in adult worms compared with other life cycle stages. We expressed in E. coli the LEL region of one of the clusters which exhibited a high frequency of transcription in female worms, and showed the purified recombinant protein (Sj-TSP-2e was recognised by 43.1% of sera obtained from confirmed schistosomiasis japonica patients. Vaccination of mice with the recombinant protein induced high levels of IgG1 and IgG2 antibodies, but no consistent protective efficacy against challenge infection was elicited in three independent trials. CONCLUSIONS/SIGNIFICANCE: The highly polymorphic nature of the Sj-TSP-2 gene at the transcriptional level may limit the value of Sj-TSP-2 as a target for future S. japonicum vaccine development.

  4. Low Sensitivity of the Formol-Ethyl Acetate Sedimentation Concentration Technique in Low-Intensity Schistosoma japonicum Infections

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    Lier, Tore; Simonsen, Gunnar S.; Wang, Tianping; Lu, Dabing; Haukland, Hanne H.; Vennervald, Birgitte J.; Johansen, Maria V.

    2009-01-01

    Background The endemic countries are in a diagnostic dilemma concerning Schistosoma japonicum with increasing difficulties in diagnosing the infected individuals. The formol-ethyl acetate sedimentation concentration technique is preferred by many clinical microbiology laboratories for the detection of parasites in stool samples. It is potentially more sensitive than the diagnostic methods traditionally used. Methodology/Principal Findings We evaluated the technique for detection of low-intensity S. japonicum infections in 106 stool samples from China and used a commercial kit, Parasep Midi Faecal Parasite Concentrator. One stool sample and one serum sample were collected from each person. As reference standard we used persons positive by indirect hemagglutination in serum and positive by Kato-Katz thick smear microscopy (three slides from a single stool), and/or the hatching test. We found the sedimentation technique to have a sensitivity of only 28.6% and specificity of 97.4%. Conclusion/Significance This study indicates that the sedimentation technique has little to offer in the diagnosis of low-intensity S. japonicum infections, at least when only a single stool sample is examined. PMID:19238192

  5. Practical application of methanol-mediated mutualistic symbiosis between Methylobacterium species and a roof greening moss, Racomitrium japonicum.

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    Akio Tani

    Full Text Available Bryophytes, or mosses, are considered the most maintenance-free materials for roof greening. Racomitrium species are most often used due to their high tolerance to desiccation. Because they grow slowly, a technology for forcing their growth is desired. We succeeded in the efficient production of R. japonicum in liquid culture. The structure of the microbial community is crucial to stabilize the culture. A culture-independent technique revealed that the cultures contain methylotrophic bacteria. Using yeast cells that fluoresce in the presence of methanol, methanol emission from the moss was confirmed, suggesting that it is an important carbon and energy source for the bacteria. We isolated Methylobacterium species from the liquid culture and studied their characteristics. The isolates were able to strongly promote the growth of some mosses including R. japonicum and seed plants, but the plant-microbe combination was important, since growth promotion was not uniform across species. One of the isolates, strain 22A, was cultivated with R. japonicum in liquid culture and in a field experiment, resulting in strong growth promotion. Mutualistic symbiosis can thus be utilized for industrial moss production.

  6. Practical application of methanol-mediated mutualistic symbiosis between Methylobacterium species and a roof greening moss, Racomitrium japonicum.

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    Tani, Akio; Takai, Yuichiro; Suzukawa, Ikko; Akita, Motomu; Murase, Haruhiko; Kimbara, Kazuhide

    2012-01-01

    Bryophytes, or mosses, are considered the most maintenance-free materials for roof greening. Racomitrium species are most often used due to their high tolerance to desiccation. Because they grow slowly, a technology for forcing their growth is desired. We succeeded in the efficient production of R. japonicum in liquid culture. The structure of the microbial community is crucial to stabilize the culture. A culture-independent technique revealed that the cultures contain methylotrophic bacteria. Using yeast cells that fluoresce in the presence of methanol, methanol emission from the moss was confirmed, suggesting that it is an important carbon and energy source for the bacteria. We isolated Methylobacterium species from the liquid culture and studied their characteristics. The isolates were able to strongly promote the growth of some mosses including R. japonicum and seed plants, but the plant-microbe combination was important, since growth promotion was not uniform across species. One of the isolates, strain 22A, was cultivated with R. japonicum in liquid culture and in a field experiment, resulting in strong growth promotion. Mutualistic symbiosis can thus be utilized for industrial moss production.

  7. Pengaruh ketiadaan inang terhadap tanggap reproduksi Trichogrammatoidea armigera Nagaraja dan Trichogramma japonicum Ashmed (Hymenoptera: Trichogrammatoidea dan implikasinya terhadap penerimaan inang

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    Anis Rohmani

    2017-02-01

    Full Text Available Trichogrammatoidea armigera and Trichogramma japonicum are polyphagous egg parastioids, that are important as natural enemies. The objective of this research was to study the effect of host deprivation on reproductive capacity of T. armigera and T. japonicum. This study consists of 8 treatments, host deprivation : 0 hour, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, and 48 hours. Host were replaced every 24 hours. Results showed there are difference with respect of how the two parasitoids respond to the treatment. Deprivation of host for 3 hours in T. armigera resulted in the increase of egg production and parasitism rate. After 12 and 24 hours of not encountering any hosts, the numbers of eggs produced drastically decreased. Treatment on T. japonicum resulted in the reaction of fecundity and parasitism rate overall. Forty eight hours of host deprivation resulted in death of both parasitoids species within 2 days. None of the parasitoids seems to produce any egg.

  8. Variation in the frequency and extent of hybridization between Leucosceptrum japonicum and L. stellipilum (Lamiaceae) in the Central Japanese Mainland.

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    Li, Yue; Maki, Masayuki

    2015-01-01

    Variations in the frequency and extent of hybridization among mixed populations located in the same contact zone provide natural laboratories for the study of extrinsic reproductive isolation maintaining species integrity. In this study, we examined the pattern of hybridization between L. japonicum and L. stellipilum among mixed populations in different localities of a contact zone. The genetic structures from three sympatric populations and six mixed populations in the hybrid zone, and five reference populations far from the contact zone, were characterized using 10 neutral nuclear microsatellite markers. Evidence from genetic distance-based clustering analysis, the frequency distribution of admixture proportion values, and the hybrid category assignment approaches indicated that the frequency and extent of hybridization varied considerably among populations in the contact zone between L. japonicum and L. stellipilum. One likely explanation is that variation in exogenous (ecological) selection among populations might contribute to differences in frequency and extent of hybridization. The present study will facilitate future research exploring the evolution of reproductive isolation between L. japonicum and L. stellipilum.

  9. High-quality permanent draft genome sequence of Bradyrhizobium sp. Th.b2, a microsymbiont of Amphicarpaea bracteata collected in Johnson City, New York.

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    Tian, Rui; Parker, Matthew; Seshadri, Rekha; Reddy, Tbk; Markowitz, Victor; Ivanova, Natalia; Pati, Amrita; Woyke, Tanja; Baeshen, Mohammed N; Baeshen, Nabih A; Kyrpides, Nikos; Reeve, Wayne

    2015-01-01

    Bradyrhizobium sp. Th.b2 is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from an effective nitrogen-fixing root nodule of Amphicarpaea bracteata collected in Johnson City, New York. Here we describe the features of Bradyrhizobium sp. Th.b2, together with high-quality permanent draft genome sequence information and annotation. The 10,118,060 high-quality draft genome is arranged in 266 scaffolds of 274 contigs, contains 9,809 protein-coding genes and 108 RNA-only encoding genes. This rhizobial genome was sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

  10. High-quality permanent draft genome sequence of Bradyrhizobium sp. strain WSM1743 - an effective microsymbiont of an Indigofera sp. growing in Australia.

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    Eshraghi, Leila; De Meyer, Sofie E; Tian, Rui; Seshadri, Rekha; Ivanova, Natalia; Pati, Amrita; Markowitz, Victor; Woyke, Tanja; Kyrpides, Nikos C; Tiwari, Ravi; Yates, Ron; Howieson, John; Reeve, Wayne

    2015-01-01

    Bradyrhizobium sp. strain WSM1743 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of an Indigofera sp. WSM1743 was isolated from a nodule recovered from the roots of an Indigofera sp. growing 20 km north of Carnarvon in Australia. It is slow growing, tolerates up to 1 % NaCl and is capable of growth at 37 °C. Here we describe the features of Bradyrhizobium sp. strain WSM1743, together with genome sequence information and its annotation. The 8,341,956 bp high-quality permanent draft genome is arranged into 163 scaffolds and 167 contigs, contains 7908 protein-coding genes and 75 RNA-only encoding genes and was sequenced as part of the Root Nodule Bacteria chapter of the Genomic Encyclopedia of Bacteria and Archaea project.

  11. The inhibitory effect against collagen-induced arthritis by Schistosoma japonicum infection is infection stage-dependent

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    Chi FengLi

    2010-06-01

    Full Text Available Abstract Background A long-term existing schistosome infection can aid in maintaining immuno-homeostasis, thus providing protection against various types of autoimmune diseases to the infected host. Such benefits have often been associated with acute or egg stage infection and with the egg-induced Th2 response. However, since schistosome infection undergoes different stages, each associated with a specific induction of Th responses, the requirements for the ability of the different stages of schistosome infection to protect against autoimmune disease has not been elucidated. The present study was designed to study whether different stages of schistosome infection offer unique protection in collagen-induced arthritis and its mechanisms. Results Arthritis susceptible strain DBA/1 male mice were infected with Schistosoma japonicum for either 2 weeks resulting in early stage infection or for 7 weeks resulting in acute or egg stage infection. Following Schistosoma japonicum infection, collagen II was administered to induce collagen-induced arthritis, an animal model for human rheumatoid arthritis. Infection by Schistosoma japonicum significantly reduced the severity and the incidence of experimental autoimmune collagen-induced arthritis. However, this beneficial effect can only be provided by a pre-established acute stage of infection but not by a pre-established early stage of the infection. The protection against collagen-induced arthritis correlated with reduced levels of anti-collagen II IgG, especially the subclass of IgG2a. Moreover, in protected mice increased levels of IL-4 were present at the time of collagen II injection together with sustained higher IL-4 levels during the course of arthritis development. In contrast, in unprotected mice minimal levels of IL-4 were present at the initial stage of collagen II challenge together with lack of IL-4 induction following Schistosoma japonicum infection. Conclusion The protective effect against

  12. Multilocus sequence analysis reveals taxonomic differences among Bradyrhizobium sp. symbionts of Lupinus albescens plants growing in arenized and non-arenized areas.

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    Granada, Camille E; Beneduzi, Anelise; Lisboa, Bruno B; Turchetto-Zolet, Andreia C; Vargas, Luciano K; Passaglia, Luciane M P

    2015-07-01

    Lupinus albescens is a leguminous plant that belongs to "New World" lupine species, which is native to southern Brazil. This Brazilian region is characterized by poor degraded soils with low organic matter and is designated as an arenized area. The symbiosis between Lupinus plants and nitrogen-fixing bacteria belonging to the Bradyrhizobium genus may help the plant establish itself in these areas. To characterize the bradyrhizobial population symbionts of L. albescens plants grown in arenized and non-arenized areas, a multilocus phylogenetic analysis allied to genetic diversity indices were conducted. Seventy-four bradyrhizobial isolates were analyzed, 38 coming from L. albescens plants growing in an arenized area and 36 from a non-arenized area. Isolates were different between arenized and non-arenized areas. Phylogenetic analysis of the 16S rRNA, dnaK, atpD, recA, glnII, rpoB, gyrB, nodA, nodB, and nodZ genes resulted in three supported clades, which were most likely to be three different new Bradyrhizobium species: one species from the arenized area and two from the non-arenized area. Estimates of genetic diversity, which decreased in arenized areas, were positively correlated with habitat variability. These results suggested that a few resistant and efficient Bradyrhizobium sp. strains were capable of forming nodules on L. albescens plants growing in an arenized area. An in vivo inoculation experiment with L. albescens plants showed that Bradyrhizobium ssp. isolated from this extreme environment were more efficient at promoting plant growth than those from the non-arenized area. This result suggested that the environment affected the selection of more efficient plant growth promoters in order to sustain plant growth. Copyright © 2015 Elsevier GmbH. All rights reserved.

  13. Influence of Schistosoma japonicum programmed cell death protein 10 on the growth and development of schistosomula.

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    Gao, Yan Ru; Huang, Wen Ling; Tang, Chun Lian; Liu, Rong; Zhao, Qin Ping; Ming, Zhen Ping; Dong, Hui Fen

    2018-01-18

    Schistosomiasis caused by Schistosoma japonicum is among the most serious endemic zoonoses in China. To study interactions between schistosomula, the pre-adult juvenile stage, and hosts, it is important to study the functions of key genes involved in schistosomula growth and development. Programmed cell death protein 10 (pcdp10) is an important apoptosis-related gene with various biological functions. This study described the molecular characterization of S. japonicum PCDP10 (SjPCDP10) and evaluated its functions in schistosomula. Real-time quantitative polymerase chain reaction (qPCR) and western blot were used to detect Sjpcdp10 mRNA and protein levels, respectively, at different developmental stages. Immunolocalization was performed to determine SjPCDP10 expression in the parasite. RNA interference (RNAi) experiments were used to assess gene functions associated with SjPCDP10 in schistosomula growth and development. Real-time qPCR revealed that Sjpcdp10 was expressed during all investigated developmental stages and upregulated during schistosomula growth and development. Histochemical localization showed that SjPCDP10 was mainly distributed in the teguments of schistosomula in all investigated stages and part of the parenchymal area of 14-, 18-, and 21-day-old schistosomula. Following Sjpcdp10 knockdown by RNAi, the lengths, widths, areas, and volumes of schistosomula were significantly lower than those in the control group. Scanning electron microscopy showed that the body surfaces of schistosomula subjected to RNAi were seriously damaged, with few tegumental spines and sensory papillae. Transmission electron microscopy indicated that the teguments of Sjpcdp10-knockdown schistosomula were incomplete, the number of layers was reduced, and the thickness decreased significantly as compared with those in the control group. Furthermore, terminal deoxynucleotidyl transferase dUTP nick-end labelling results showed that the rate of apoptosis in Sjpcdp10-knockdown

  14. MALDI-TOF mass spectrometry as a tool for differentiation of Bradyrhizobium species: application to the identification of Lupinus nodulating strains.

    Science.gov (United States)

    Sánchez-Juanes, Fernando; Ferreira, Laura; Alonso de la Vega, Pablo; Valverde, Angel; Barrios, Milagros León; Rivas, Raúl; Mateos, Pedro F; Martínez-Molina, Eustoquio; González-Buitrago, José Manuel; Trujillo, Martha E; Velázquez, Encarna

    2013-12-01

    Genus Bradyrhizobium includes slow growing bacteria able to nodulate different legumes as well as species isolated from plant tumours. The slow growth presented by the members of this genus and the phylogenetic closeness of most of its species difficults their identification. In the present work we applied for the first time Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) to the analysis of Bradyrhizobium species after the extension of MALDI Biotyper 2.0 database with the currently valid species of this genus. With this methodology it was possible to identify strains belonging to phylogenetically closely related species of genus Bradyrhizobium allowing the discrimination among species with rrs gene identities higher than 99%. The application of MALDI-TOF MS to strains isolated from nodules of different Lupinus species in diverse geographical locations allowed their correct identification when comparing with the results of rrs gene and ITS analyses. The nodulation of Lupinus gredensis, an endemic species of the west of Spain, by B. canariense supports the European origin of this species. Copyright © 2013. Published by Elsevier GmbH.

  15. SjTat-TPI facilitates adaptive T-cell responses and reduces hepatic pathology during Schistosoma japonicum infection in BALB/c mice.

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    Zhang, Wenyue; Luo, Xiaofeng; Zhang, Fan; Zhu, Yuxiao; Yang, Bingya; Hou, Min; Xu, Zhipeng; Yu, Chuanxin; Chen, Yingying; Chen, Lin; Ji, Minjun

    2015-12-30

    Schistosomiasis is a kind of parasitic zoonoses which causes serious damage to public health and social development. China is one of the countries most affected by Schistosoma japonicum and an effective vaccine is still needed. In this study, we adopted Tat-mediated protein transduction technology to investigate the impact of different antigen presented approaches on host's immune response and the potential protection against Schistosoma japonicum infection. We successfully constructed the recombinant S. japonicum triosephosphate isomerase, Tat-TPI, as a vaccine candidate. Whether injected with Tat-TPI in foot pad or vaccinated with Tat-TPI in the back subcutaneously for three times, the draining popliteal lymph nodes and spleen both developed a stronger CD8(+)T response (Tc1) in mice. Not only that, but it also helped CD4(+)T cells to produce more IFN-γ than TPI immunisation. In addition, it could boost IgG production, especially IgG1 subclass. Most importantly, Tat-TPI immunisation led to the significant smaller area of a single egg granuloma in the livers as compared with TPI-vaccinated or control groups. However, the anti-infection efficiency induced by Tat-TPI was still restricted. This study indicated that immunisation with Tat-fused TPI could contribute to enhance CD4(+)T-cell response and decrease hepatic egg granulomatous area after S. japonicum infection though it did not achieve our expected protection against Schistosoma japonicum infection. The optimal vaccine strategy warrants further research.

  16. Genetic diversity and structure of Schistosoma japonicum within two marshland villages of Anhui, China, prior to schistosome transmission control and elimination.

    Science.gov (United States)

    Ding, Huan; Lu, Da-Bing; Gao, Yu-Meng; Deng, Yao; Li, Ying

    2017-02-01

    Schistosomiasis is caused by the genus Schistosoma and affected more than 250 million people worldwide. Schistosoma japonicum was once seriously endemic in China and nearly 60 years of efforts has seen great success in disease control. However, due to its zoonotic nature and complex life cycle, the schistosomiasis transmission control and final elimination would require, besides an intersectoral approach, deep understanding of population genetics of the parasite. We therefore performed a snail survey in two marshland villages of Anhui province of China and collected S. japonicum cercariae from infected snails. By using the recent developed microsatellite panel comprising seven loci, we genotyped the sampled parasites and analyzed the population genetic diversity and structure. The results showed much lower infection prevalence of S. japonicum in snails and low infected snail density in either marshland village. Through population genetic analyses, a considerable genetic diversity of parasites was revealed, whereas a small number of clusters were inferred and the sign of bottleneck effect was detected in each village. For the first time in S. japonicum in two villages, we provided estimates of effective population sizes with two different approaches. The results indicated that the parasite in two villages could eventually be eradicated with the ongoing integral control measures, but with potential risk of reinvasion of immigrant parasites through the Yangtze River. Such would be of great importance in assessment of the effects of ongoing control measures and prediction of the transmission capability for S. japonicum, thus guiding decisions on the choice of further control work.

  17. [Effect of ICOS signaling on CD154/CD40 expressions in mice infected with Schistosoma japonicum].

    Science.gov (United States)

    Wang, Yu; Cai, Ru; Xia, Chao-ming

    2015-08-01

    To explore the effect of ICOS signaling on the CD154/CD40 expressions and immunopathology in mice infected with Schistosoma japonicum. ICOS transgenic (ICOS-Tg) mice and wildtype FVB/NJ mice were used as experimental schistosomiasis models. The expressions of CD154 and CD40 on splenocytes and on inflammatory cells around granulomatous infiltration of the liver in the mice infected with S. japonicuin were detected by flow cytometry and im- munohistochemical staining. HE staining was applied to observe the changes on the granulomatous of the mice liver. Compared with the wildtype FVB/NJ mice, the expressions of CD154 on CD4 T splenocytes and of CD40 on CD19' B splenocytes in the ICOS-Tg mice significantly increased in 12 and 16 weeks post-infection (all P CD154 on inflammatory cells around granulomatous infiltration in the liver of the ICOS-Tg mice were significantly higher than those of the wildtype FVB/NJ mice in 7, 12, 16 and 20 weeks post-infection (all P CD154/CD40 expressions, and may play an important role in the hepatic egg granuloma formation of schistosomiasis.

  18. Chemical constituents from the fruits of Ligustrum japonicum and their inhibitory effects on T cell activation.

    Science.gov (United States)

    Ngo, Quynh-Mai Thi; Lee, Hyun-Su; Nguyen, Van Thu; Kim, Jeong Ah; Woo, Mi Hee; Min, Byung Sun

    2017-09-01

    A previously undescribed nor-dammarane, 3β,20,23-trihydroxy-24,25,26,27-tetranordammarane; three previously undescribed secoiridoid glycosides, ligujaponosides A-B, and iso-oleonuzhenide; and twenty three known compounds, were isolated from the fruits of Ligustrum japonicum Thunb. Their chemical structures were elucidated by extensive spectroscopic analyses, including 1D and 2D NMR, and HRMS. The isolated compounds were screened for immunosuppressive effects on T activated cells by evaluating interleukin-2 (IL-2) production. Among them, sesamin inhibited IL-2 production in Jurkat T cells with an IC50 value of 38 ± 2 μM. In addition, sesamin inhibited the phosphorylation of extracellular signal-regulated protein kinase (ERK), a member of the mitogen-activated protein kinase (MAPK) family, in phorbol 12-myristate 13-acetate (PMA)/A23187-stimulated T cells. Therefore, sesamin was demonstrated to inhibit T cell activation via regulation of MAPK phosphorylation pathway. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Helminth Protein Vaccine Induced Follicular T Helper Cell for Enhancement of Humoral Immunity against Schistosoma japonicum

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    Jingyao Zhang

    2013-01-01

    Full Text Available Protein vaccines combined with adjuvants have been widely used to induce immune responses, especially the humoral immune response, against molecular targets including parasites. Follicular T helper (Tfh cells are the specialized providers of B-cell help, however, the induction of Tfh cells in protein vaccination has been rarely studied. Here, we report that the Schistosoma japonicum recombinant protein (SjGST-32 combined with tacrolimus (FK506 augmented the induction of Tfh cells, which expressed the canonical markers CXCR5, BCL6, and IL-21, and enhanced the humoral immune responses in BALB/c mice. Furthermore, the expression of IL-21R on germinal center (GC B cells and memory B cells increased in immunized mice, which indicated that IL-21 from the induced Tfh cells interacted with IL-21R for activation of B cells and maintenance of long-lived humoral immunity. Our results suggest that helminth protein vaccine combined with FK506 induces Tfh cell for stimulating humoral immune responses and inducing long-lived humoral immunity.

  20. Magnetic Resonance Spectroscopy for Evaluating Portal-Systemic Encephalopathy in Patients with Chronic Hepatic Schistosomiasis Japonicum.

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    Li, Ying; Mei, Lihong; Qiang, Jinwei; Ju, Shuai; Zhao, Shuhui

    2016-12-01

    Portal-systemic encephalopathy (PSE) is classified as type B hepatic encephalopathy. Portal-systemic shunting rather than liver dysfunction is the main cause of PSE in chronic hepatic schistosomiasis japonicum (HSJ) patients. Owing to lack of detectable evidence of intrinsic liver disease, chronic HSJ patients with PSE are frequently clinically undetected or misdiagnosed, especially chronic HSJ patients with covert PSE (subclinical encephalopathy). In this study, we investigated whether magnetic resonance spectroscopy (MRS) could be a useful tool for diagnosing PSE in chronic HSJ patients. Magnetic resonance (MR) T1-weighted imaging, diffusion-weighted imaging, and MRS were performed in 41 chronic HSJ patients with suspected PSE and in 21 age-matched controls. The T1 signal intensity index (T1SI) and apparent diffusion coefficient (ADC) value were obtained in the Globus pallidus. Liver function was also investigated via serum ammonia and liver function tests. Higher T1SI and ADC values, increased lactate and glutamine levels, and decreased myo-inositol were found in the bilateral Globus pallidus in chronic HSJ patients with PSE. No significantly abnormal serum ammonia or liver function tests were observed in chronic HSJ patients with PSE. On the basis of these findings, we propose a diagnostic procedure for PSE in chronic HSJ patients. This study reveals that MRS can be useful for diagnosing PSE in chronic HSJ patients.

  1. Chronic Schistosoma japonicum infection reduces immune response to vaccine against hepatitis B in mice.

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    Lin Chen

    Full Text Available BACKGROUND: Hepatitis B and schistosomiasis are most prevalent in Africa and Asia, and co-infections of both are frequent in these areas. The immunomodulation reported to be induced by schistosome infections might restrict immune control of hepatitis B virus (HBV leading to more severe viral infection. Vaccination is the most effective measure to control and prevent HBV infection, but there is evidence for a reduced immune response to the vaccine in patients with chronic schistosomiasis japonica. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we demonstrate in a mouse model that a chronic Schistosoma japonicum infection can inhibit the immune response to hepatitis B vaccine (HBV vaccine and lead to lower production of anti-HBs antibodies, interferon-γ (IFN-γ and interleukin-2 (IL-2. After deworming with Praziquantel (PZQ, the level of anti-HBs antibodies gradually increased and the Th2-biased profile slowly tapered. At 16 weeks after deworming, the levels of anti-HBs antibodies and Th1/Th2 cytokines returned to the normal levels. CONCLUSIONS/SIGNIFICANCE: The results suggest that the preexisting Th2-dominated immune profile in the host infected with the parasite may down-regulate levels of anti-HBs antibodies and Th1 cytokines. To improve the efficacy of HBV vaccination in schistosome infected humans it may be valuable to treat them with praziquantel (PZQ some time prior to HBV vaccination.

  2. Antischistosomal activity of hederacochiside C against Schistosoma japonicum harbored in experimentally infected animals.

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    Kang, Nai-Xin; Zhu, Yuan-Jian; Zhao, Jian-Ping; Zhu, Wei-Feng; Liu, Yan-Li; Xu, Qiong-Ming; Zhuge, Hong-Xiang; Khan, Ikhlas A; Yang, Shi-Lin

    2017-04-01

    The present study was undertaken to investigate whether hederacochiside C (HSC) possesses antischistosomal effects and anti-inflammatory response activities in Schistosoma japonicum-infected mice. Different concentrations of HSC were administrated to the mice infected by schistosomula or adult worm by intravenous injection twice a day for five consecutive days. The total worm burden, female worm burden, and the egg burden in liver of mice treated with 400 mg/kg HSC were fewer than those in non-treated ones. Murine immune responses following HSC treatment were investigated using enzyme-linked immunosorbent assays (ELISA). Our results indicated that 200 mg/kg HSC could reduce the expression of IgG, tumor necrosis factor (TNF)-α, interleukin (IL)-4 and IL-17 in comparison to infected group, exhibiting best immunomodulatory effects. In addition, scanning electron microscopical examination revealed that male worms treated with HSC lost their normal surface architecture since its surface showed extensive swelling, erosion, and peeling in tegumental regions. Remarkable amelioration was noticed in histopathological investigations, and 200 mg/kg HSC treatment could reduce the size of granulomatous inflammatory infiltrations in the liver which was reflected in nearly normalization of liver architecture. These results suggested that HSC had potential antischistosomal activity and provided a basis for subsequent experimental.

  3. Tissue specific profiling of females of Schistosoma japonicum by integrated laser microdissection microscopy and microarray analysis.

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    Geoffrey N Gobert

    2009-06-01

    Full Text Available The functions of many schistosome gene products remain to be characterized. A major step towards elucidating function of these genes would be in defining their sites of expression. This goal is rendered difficult to achieve by the generally small size of the parasites and the lack of a body cavity, which precludes analysis of transcriptional profiles of the tissues in isolation.Here, we describe a combined laser microdissection microscopy (LMM and microarray analysis approach to expedite tissue specific profiling and gene atlasing for tissues of adult female Schistosoma japonicum. This approach helps to solve the gene characterization "bottle-neck" brought about by acoelomy and the size of these parasites. Complementary RNA obtained after isolation from gastrodermis (parasite gut mucosa, vitelline glands and ovary by LMM were subjected to microarray analyses, resulting in identification of 147 genes upregulated in the gastrodermis, 4,149 genes in the ovary and 2,553 in the vitellaria.This work will help to shed light on the molecular pathobiology of this debilitating human parasite and aid in the discovery of new targets for the development of anti-schistosome vaccines and drugs.

  4. A Microtus fortis protein, serum albumin, is a novel inhibitor of Schistosoma japonicum schistosomula

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    Rong Li

    2013-11-01

    Full Text Available Schistosomiasis is an endemic parasite disease and praziquantel is the only drug currently in use to control this disease. Experimental and epidemiological evidence strongly suggests that Microtus fortis ( Mf is a naturally resistant vertebrate host of Schistosoma japonicum . In the present study, we found that Mf serum albumin ( Mf -albumin and the conditioned medium of pcDNA3.1- Mf -albumin caused 46.2% and 38.7% schistosomula death rates in 96 h, respectively, which were significantly higher than that of the negative control (p < 0.05. We also found that mice injected with Mf -albumin had a 43.5% reduction in worm burden and a 48.1% reduction in liver eggs per gram (p < 0.05 in comparison to the control animals. To characterise the mechanisms involved in clearance, schistosomula were incubated with fluorescein isothiocyanate-labelled Mf -albumin and fluorescent enrichment effects were found in the gut lumen of schistosomula after 48 h of incubation. Next, digestive tract excretions from schistosomula were collected and the sensitivity of Mf -albumin to digestive tract excretions was evaluated. The results indicated that schistosomula digestive tract excretions showed indigestibility of Mf -albumin. The death of schistosomula could be partially attributed to the lack of digestion of Mf -albumin by digestive tract excretions during the development of the schistosomula stage. Therefore, these data indicate the potential of Mf -albumin as one of the major selective forces for schistosomiasis.

  5. Influência do pH do meio de cultivo e da turfa no comportamento de estirpes de Bradyrhizobium

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    D. L. Miguel

    2001-12-01

    Full Text Available A acidez dos solos representa um problema que afeta grandes áreas agrícolas pelo mundo, principalmente nos trópicos, onde fósforo e nitrogênio também são limitantes. No caso do nitrogênio, a fixação biológica torna-se uma das alternativas mais viáveis do ponto de vista ecológico e econômico, por diminuir o uso e o impacto causado pelos fertilizantes nitrogenados. Neste trabalho, foram realizados dois experimentos in vitro e um em casa de vegetação com quatro estirpes de Bradyrhizobium (Br 4406, Br 29, SEMIA 587 e INPA 03-11B, no Departamento de Ciência do Solo (UFLA, de julho de 1998 a julho de 1999, para verificar o efeito de três valores de pH (5,0; 6,0; 6,9 no crescimento destas em meio de cultura YM, na sua simbiose com soja, assim como na sua sobrevivência em inoculantes produzidos com turfa. No primeiro experimento, as estirpes de Bradyrhizobium tiveram um comportamento diferenciado em meio líquido, obtendo melhor desempenho em pH 6,0, tanto em número de unidades formadoras de colônias quanto em produção de exopolissacarídeos. +No segundo experimento, o número de nódulos, a atividade da nitrogenase (Nase, as massas secas de nódulos, raízes e parte aérea de plantas de soja, de modo geral, não foram influenciados pelos valores de pH de cultivo das estirpes no inoculante. Entretanto, a estirpe INPA 03 - 11B mostrou-se efetiva, apresentando número de nódulos e atividade de Nase semelhantes aos de Br 29 e SEMIA 587, que são recomendadas como estirpes inoculantes, devendo, assim, ser indicada para testes de eficiência em campo. No terceiro experimento, com exceção da Br 29, que atingiu maior sobrevivência de células em pH 6,0, as outras estirpes tiveram sobrevivência semelhante neste valor de pH e em pH 6,9. O melhor desempenho das estirpes de Bradyrhizobium em pH 6,0 no meio de cultura e em turfa demonstrou a possibilidade do uso de inoculantes corrigidos para esse valor de pH, como modo de pr

  6. Homology-based annotation of non-coding RNAs in the genomes of Schistosoma mansoni and Schistosoma japonicum

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    Santana Clara

    2009-10-01

    Full Text Available Abstract Background Schistosomes are trematode parasites of the phylum Platyhelminthes. They are considered the most important of the human helminth parasites in terms of morbidity and mortality. Draft genome sequences are now available for Schistosoma mansoni and Schistosoma japonicum. Non-coding RNA (ncRNA plays a crucial role in gene expression regulation, cellular function and defense, homeostasis, and pathogenesis. The genome-wide annotation of ncRNAs is a non-trivial task unless well-annotated genomes of closely related species are already available. Results A homology search for structured ncRNA in the genome of S. mansoni resulted in 23 types of ncRNAs with conserved primary and secondary structure. Among these, we identified rRNA, snRNA, SL RNA, SRP, tRNAs and RNase P, and also possibly MRP and 7SK RNAs. In addition, we confirmed five miRNAs that have recently been reported in S. japonicum and found two additional homologs of known miRNAs. The tRNA complement of S. mansoni is comparable to that of the free-living planarian Schmidtea mediterranea, although for some amino acids differences of more than a factor of two are observed: Leu, Ser, and His are overrepresented, while Cys, Meth, and Ile are underrepresented in S. mansoni. On the other hand, the number of tRNAs in the genome of S. japonicum is reduced by more than a factor of four. Both schistosomes have a complete set of minor spliceosomal snRNAs. Several ncRNAs that are expected to exist in the S. mansoni genome were not found, among them the telomerase RNA, vault RNAs, and Y RNAs. Conclusion The ncRNA sequences and structures presented here represent the most complete dataset of ncRNA from any lophotrochozoan reported so far. This data set provides an important reference for further analysis of the genomes of schistosomes and indeed eukaryotic genomes at large.

  7. Real-time PCR demonstrates high prevalence of Schistosoma japonicum in the Philippines: implications for surveillance and control.

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    Catherine A Gordon

    2015-01-01

    Full Text Available The Philippines has a population of approximately 103 million people, of which 6.7 million live in schistosomiasis-endemic areas with 1.8 million people being at risk of infection with Schistosoma japonicum. Although the country-wide prevalence of schistosomiasis japonica in the Philippines is relatively low, the prevalence of schistosomiasis can be high, approaching 65% in some endemic areas. Of the currently available microscopy-based diagnostic techniques for detecting schistosome infections in the Philippines and elsewhere, most exhibit varying diagnostic performances, with the Kato-Katz (KK method having particularly poor sensitivity for detecting low intensity infections. This suggests that the actual prevalence of schistosomiasis japonica may be much higher than previous reports have indicated.Six barangay (villages were selected to determine the prevalence of S. japonicum in humans in the municipality of Palapag, Northern Samar. Fecal samples were collected from 560 humans and examined by the KK method and a validated real-time PCR (qPCR assay. A high S. japonicum prevalence (90.2% was revealed using qPCR whereas the KK method indicated a lower prevalence (22.9%. The geometric mean eggs per gram (GMEPG determined by the qPCR was 36.5 and 11.5 by the KK. These results, particularly those obtained by the qPCR, indicate that the prevalence of schistosomiasis in this region of the Philippines is much higher than historically reported.Despite being more expensive, qPCR can complement the KK procedure, particularly for surveillance and monitoring of areas where extensive schistosomiasis control has led to low prevalence and intensity infections and where schistosomiasis elimination is on the horizon, as for example in southern China.

  8. The RIO protein kinase-encoding gene Sj-riok-2 is involved in key reproductive processes in Schistosoma japonicum.

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    Zhao, Lu; He, Xin; Grevelding, Christoph G; Ye, Qing; Li, Ying; Gasser, Robin B; Dissous, Colette; Mughal, Mudassar N; Zhou, Yan-Qin; Zhao, Jun-Long; Hu, Min

    2017-12-12

    Schistosomiasis is one of the most prevalent parasitic diseases worldwide and is caused by parasitic trematodes of the genus Schistosoma. The pathogenesis of schistosomiasis is caused by eggs whose production is the consequence of the pairing of schistosomes and the subsequent sexual maturation of the female. Previous studies have demonstrated that protein kinases are involved in processes leading to the male-induced differentiation of the female gonads, ovary and vitellarium. Right open reading frame protein kinase 2 (RIOK-2) is a member of the atypical kinase family and shown in other organisms to be responsible for ribosomal RNA biogenesis and cell-cycle progression, as well as involves in nematode development. However, nothing is known about its functions in any trematode including schistosome. We isolated and characterized the riok-2 gene from S. japonicum, and detected the transcriptional profiles of Sj-riok-2 by using real-time PCR and in situ hybridization. RNAi-mediated knockdown of Sj-riok-2 was performed, mitotic activities were detected by EdU incorporation assay and morphological changes on organs were observed by confocal laser scanning microscope (CLSM). In silico analyses of the amino acid sequence of Sj-RIOK-2 revealed typical features of this class of kinases including a winged helix (wHTH) domain and a RIO kinase domain. Sj-riok-2 is transcribed in different developmental stages of S. japonicum, with a higher abundance in adult females and eggs. Localization studies showed that Sj-riok-2 was mainly transcribed in female reproductive organs. Experiments with adult schistosomes in vitro demonstrated that the transcriptional level of Sj-riok-2 was affected by pairing. Knocking down Sj-riok-2 by RNAi reduced cell proliferation in the vitellarium and caused the increased amount of mature oocytes in ovary and an accumulation of eggs within the uterus. Sj-riok-2 is involved in the reproductive development and maturation of female S. japonicum. Our

  9. Construction and simulation of the Bradyrhizobium diazoefficiens USDA110 metabolic network: a comparison between free-living and symbiotic states.

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    Yang, Yi; Hu, Xiao-Pan; Ma, Bin-Guang

    2017-02-28

    Bradyrhizobium diazoefficiens is a rhizobium able to convert atmospheric nitrogen into ammonium by establishing mutualistic symbiosis with soybean. It has been recognized as an important parent strain for microbial agents and is widely applied in agricultural and environmental fields. In order to study the metabolic properties of symbiotic nitrogen fixation and the differences between a free-living cell and a symbiotic bacteroid, a genome-scale metabolic network of B. diazoefficiens USDA110 was constructed and analyzed. The metabolic network, iYY1101, contains 1031 reactions, 661 metabolites, and 1101 genes in total. Metabolic models reflecting free-living and symbiotic states were determined by defining the corresponding objective functions and substrate input sets, and were further constrained by high-throughput transcriptomic and proteomic data. Constraint-based flux analysis was used to compare the metabolic capacities and the effects on the metabolic targets of genes and reactions between the two physiological states. The results showed that a free-living rhizobium possesses a steady state flux distribution for sustaining a complex supply of biomass precursors while a symbiotic bacteroid maintains a relatively condensed one adapted to nitrogen-fixation. Our metabolic models may serve as a promising platform for better understanding the symbiotic nitrogen fixation of this species.

  10. Differing courses of genetic evolution of Bradyrhizobium inoculants as revealed by long-term molecular tracing in Acacia mangium plantations.

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    Perrineau, M M; Le Roux, C; Galiana, A; Faye, A; Duponnois, R; Goh, D; Prin, Y; Béna, G

    2014-09-01

    Introducing nitrogen-fixing bacteria as an inoculum in association with legume crops is a common practice in agriculture. However, the question of the evolution of these introduced microorganisms remains crucial, both in terms of microbial ecology and agronomy. We explored this question by analyzing the genetic and symbiotic evolution of two Bradyrhizobium strains inoculated on Acacia mangium in Malaysia and Senegal 15 and 5 years, respectively, after their introduction. Based on typing of several loci, we showed that these two strains, although closely related and originally sampled in Australia, evolved differently. One strain was recovered in soil with the same five loci as the original isolate, whereas the symbiotic cluster of the other strain was detected with no trace of the three housekeeping genes of the original inoculum. Moreover, the nitrogen fixation efficiency was variable among these isolates (either recombinant or not), with significantly high, low, or similar efficiencies compared to the two original strains and no significant difference between recombinant and nonrecombinant isolates. These data suggested that 15 years after their introduction, nitrogen-fixing bacteria remain in the soil but that closely related inoculant strains may not evolve in the same way, either genetically or symbiotically. In a context of increasing agronomical use of microbial inoculants (for biological control, nitrogen fixation, or plant growth promotion), this result feeds the debate on the consequences associated with such practices. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  11. NODULATION OF YELLOW LUPIN (Lupinus luteus L. DEPENDING ON THE FORECROP, SEED INOCULATION WITH Bradyrhizobium lupini AND GENISTEIN

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    Janusz PRUSIŃSKI

    2012-12-01

    Full Text Available A strict two-factor field ‘Mister’ yellow lupin experiment was made in a split-plot design in poor rye complex soil (soil valuation class IVb at the Experiment Station of the Faculty of Agriculture and Biotechnology, the University of Technology and Life Sciences at Mochełek over 2008-2011. The experiment design covered 2 factors: growing yellow lupin after intercrops of ‘Bardena’ white mustard and ‘Pastar’ winter rye as well as seed inoculation with Bradyrhizobium lupini with or without genistein added. Growing intercrops in the poor rye complex soil considerably decreased neither the Nmin content in early spring nor the size of nodulation in yellow lupin. After lupin harvest, slightly more Nmin was noted for the treatments where intercrops were applied, especially rye in the 0-30 cm layer. The weather pattern, especially moisture conditions, pointed to a special sensitivity of nodulation to water deficit or excess in the rhizosphere. The method of preparing seeds for sowing did not play a greater role in developing the Nmin content in both soil profile layers analysed. White mustard showed a negative effect on the dry weight of nodules and the total N content in the plants and the seeds of yellow lupin as well as created the least favourable conditions for the survival and the capacity for symbiosis by symbiotic bacteria remaining in soil after lupin harvest. The state of yellow lupin plant nitrogen nutrition did not depend on the experimental factors.

  12. Metabolomic Profiling of Bradyrhizobium diazoefficiens-Induced Root Nodules Reveals Both Host Plant-Specific and Developmental Signatures

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    Martina Lardi

    2016-05-01

    Full Text Available Bradyrhizobium diazoefficiens is a nitrogen-fixing endosymbiont, which can grow inside root-nodule cells of the agriculturally important soybean and other host plants. Our previous studies described B. diazoefficiens host-specific global expression changes occurring during legume infection at the transcript and protein level. In order to further characterize nodule metabolism, we here determine by flow injection–time-of-flight mass spectrometry analysis the metabolome of (i nodules and roots from four different B. diazoefficiens host plants; (ii soybean nodules harvested at different time points during nodule development; and (iii soybean nodules infected by two strains mutated in key genes for nitrogen fixation, respectively. Ribose (soybean, tartaric acid (mungbean, hydroxybutanoyloxybutanoate (siratro and catechol (cowpea were among the metabolites found to be specifically elevated in one of the respective host plants. While the level of C4-dicarboxylic acids decreased during soybean nodule development, we observed an accumulation of trehalose-phosphate at 21 days post infection (dpi. Moreover, nodules from non-nitrogen-fixing bacteroids (nifA and nifH mutants showed specific metabolic alterations; these were also supported by independent transcriptomics data. The alterations included signs of nitrogen limitation in both mutants, and an increased level of a phytoalexin in nodules induced by the nifA mutant, suggesting that the tissue of these nodules exhibits defense and stress reactions.

  13. Assessment of the diagnostic efficacy of enolase as an indication of active infection of Schistosoma japonicum.

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    Gao, Hong; Xiao, Di; Song, Lijun; Zhang, Wei; Shen, Shuang; Yin, Xuren; Wang, Jie; Ke, Xuedan; Yu, Chuanxin; Zhang, Jianzhong

    2016-01-01

    Schistosomiasis is a common zoonoses affecting humans. The atypical clinical symptoms, low morbidity, and low degree of infection impede diagnosis and assessment of epidemics. Detecting circulating antigens from adult worms in patients' body fluids should be diagnostically superior to examining eggs in feces. Herein, the excretory-secretory proteins of adult worms were analyzed by using 2-D protein electrophoresis and mass spectrometry. The Schistosoma japonicum enolase (Sj enolase) was identified as the most abundant excretory-secretory antigen. Purified recombinant Sj enolase was prepared, and specific monoclonal and polyclonal antibodies were raised against it. A sandwich enzyme-linked immunoassay (sandwich ELISA) was established that used the monoclonal antibody as a capture antibody and the polyclonal antibody as a detection antibody. The linear detection range was 0.7-1000 ng/ml (minimum 700 pg/ml). Sj enolase could be detected in the sera of infected rabbits and disappeared rapidly postpraziquantel treatment. The sensitivity and specificity of this sandwich ELISA to detect field serum samples of schistosomiasis were 84.61 and 95.83 %, respectively. The cross-reaction rates for clonorchiasis and paragonimiasis were 3.33 and 5 %, respectively. This ELISA assay was used to test 45 matching sera of schistosomiasis patients before treatment and at 3, 6, 9, and 12 months posttreatment. Among the sera, 88.89 % were positive before treatment. At 3, 6, 9, and 12 months postpraziquantel treatment, 93.33, 97.78, 100, and 100 % tested negative, respectively. Therefore, Sj enolase can be used to indicate active Schistosoma infection, and detecting serum Sj enolase is important for diagnosis and evaluating treatment effect.

  14. Development of oral and branchial muscles in lancelet larvae of Branchiostoma japonicum.

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    Yasui, Kinya; Kaji, Takao; Morov, Arseniy R; Yonemura, Shigenobu

    2014-04-01

    The perforated pharynx has generally been regarded as a shared characteristic of chordates. However, there still remains phylogenetic ambiguity between the cilia-driven system in invertebrate chordates and the muscle-driven system in vertebrates. Giant larvae of the genus Asymmetron were reported to develop an orobranchial musculature similar to that of vertebrates more than 100 years ago. This discovery might represent an evolutionary link for the chordate branchial system, but few investigations of the lancelet orobranchial musculature have been completed since. We studied staged larvae of a Japanese population of Branchiostoma japonicum to characterize the developmental property of the orobranchial musculature. The larval mouth and the unpaired primary gills develop well-organized muscles. These muscles function only as obturators of the openings without antagonistic system. As the larval mouth enlarged posteriorly to the level of the ninth myomere, the oral musculature was fortified accordingly without segmental patterning. In contrast, the iterated branchial muscles coincided with the dorsal myomeric pattern before metamorphosis, but the pharynx was remodeled dynamically irrespective of the myomeric pattern during metamorphosis. The orobranchial musculature disappeared completely during metamorphosis, and adult muscles in the oral hood and velum, as well as on the pterygial coeloms developed independently. The lancelet orobranchial musculature is apparently a larval adaptation to prevent harmful intake. However, vestigial muscles appeared transiently with the secondary gill formation suggest a bilateral ancestral state of muscular gills, and a segmental pattern of developing branchial muscles without neural crest and placodal contributions is suggestive of a precursor of vertebrate branchiomeric pattern. Copyright © 2013 Wiley Periodicals, Inc.

  15. Evidence for at least six Hox clusters in the Japanese lamprey (Lethenteron japonicum)

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    Mehta, Tarang K.; Ravi, Vydianathan; Yamasaki, Shinichi; Lee, Alison P.; Lian, Michelle M.; Tay, Boon-Hui; Tohari, Sumanty; Yanai, Seiji; Tay, Alice; Brenner, Sydney; Venkatesh, Byrappa

    2013-01-01

    Cyclostomes, comprising jawless vertebrates such as lampreys and hagfishes, are the sister group of living jawed vertebrates (gnathostomes) and hence an important group for understanding the origin and diversity of vertebrates. In vertebrates and other metazoans, Hox genes determine cell fate along the anteroposterior axis of embryos and are implicated in driving morphological diversity. Invertebrates contain a single Hox cluster (either intact or fragmented), whereas elephant shark, coelacanth, and tetrapods contain four Hox clusters owing to two rounds of whole-genome duplication (“1R” and “2R”) during early vertebrate evolution. By contrast, most teleost fishes contain up to eight Hox clusters because of an additional “teleost-specific” genome duplication event. By sequencing bacterial artificial chromosome (BAC) clones and the whole genome, here we provide evidence for at least six Hox clusters in the Japanese lamprey (Lethenteron japonicum). This suggests that the lamprey lineage has experienced an additional genome duplication after 1R and 2R. The relative age of lamprey and human paralogs supports this hypothesis. Compared with gnathostome Hox clusters, lamprey Hox clusters are unusually large. Several conserved noncoding elements (CNEs) were predicted in the Hox clusters of lamprey, elephant shark, and human. Transgenic zebrafish assay indicated the potential of CNEs to function as enhancers. Interestingly, CNEs in individual lamprey Hox clusters are frequently conserved in multiple Hox clusters in elephant shark and human, implying a many-to-many orthology relationship between lamprey and gnathostome Hox clusters. Such a relationship suggests that the first two rounds of genome duplication may have occurred independently in the lamprey and gnathostome lineages. PMID:24043829

  16. Characterization of the Runx Gene Family in a Jawless Vertebrate, the Japanese Lamprey (Lethenteron japonicum)

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    Nah, Giselle Sek Suan; Tay, Boon-Hui; Brenner, Sydney; Osato, Motomi; Venkatesh, Byrappa

    2014-01-01

    The cyclostomes (jawless vertebrates), comprising lampreys and hagfishes, are the sister group of jawed vertebrates (gnathostomes) and are hence an important group for the study of vertebrate evolution. In mammals, three Runx genes, Runx1, Runx2 and Runx3, encode transcription factors that are essential for cell proliferation and differentiation in major developmental pathways such as haematopoiesis, skeletogenesis and neurogenesis and are frequently associated with diseases. We describe here the characterization of Runx gene family members from a cyclostome, the Japanese lamprey (Lethenteron japonicum). The Japanese lamprey contains three Runx genes, RunxA, RunxB, and RunxC. However, phylogenetic and synteny analyses suggest that they are not one-to-one orthologs of gnathostome Runx1, Runx2 and Runx3. The major protein domains and motifs found in gnathostome Runx proteins are highly conserved in the lamprey Runx proteins. Although all gnathostome Runx genes each contain two alternative promoters, P1 (distal) and P2 (proximal), only lamprey RunxB possesses the alternative promoters; lamprey RunxA and RunxC contain only P2 and P1 promoter, respectively. Furthermore, the three lamprey Runx genes give rise to fewer alternative isoforms than the three gnathostome Runx genes. The promoters of the lamprey Runx genes lack the tandem Runx-binding motifs that are highly conserved among the P1 promoters of gnathostome Runx1, Runx2 and Runx3 genes; instead these promoters contain dispersed single Runx-binding motifs. The 3′UTR of lamprey RunxB contains binding sites for miR-27 and miR-130b/301ab, which are conserved in mammalian Runx1 and Runx3, respectively. Overall, the Runx genes in lamprey seem to have experienced a different evolutionary trajectory from that of gnathostome Runx genes which are highly conserved all the way from cartilaginous fishes to mammals. PMID:25405766

  17. Characterization of the Runx gene family in a jawless vertebrate, the Japanese lamprey (Lethenteron japonicum.

    Directory of Open Access Journals (Sweden)

    Giselle Sek Suan Nah

    Full Text Available The cyclostomes (jawless vertebrates, comprising lampreys and hagfishes, are the sister group of jawed vertebrates (gnathostomes and are hence an important group for the study of vertebrate evolution. In mammals, three Runx genes, Runx1, Runx2 and Runx3, encode transcription factors that are essential for cell proliferation and differentiation in major developmental pathways such as haematopoiesis, skeletogenesis and neurogenesis and are frequently associated with diseases. We describe here the characterization of Runx gene family members from a cyclostome, the Japanese lamprey (Lethenteron japonicum. The Japanese lamprey contains three Runx genes, RunxA, RunxB, and RunxC. However, phylogenetic and synteny analyses suggest that they are not one-to-one orthologs of gnathostome Runx1, Runx2 and Runx3. The major protein domains and motifs found in gnathostome Runx proteins are highly conserved in the lamprey Runx proteins. Although all gnathostome Runx genes each contain two alternative promoters, P1 (distal and P2 (proximal, only lamprey RunxB possesses the alternative promoters; lamprey RunxA and RunxC contain only P2 and P1 promoter, respectively. Furthermore, the three lamprey Runx genes give rise to fewer alternative isoforms than the three gnathostome Runx genes. The promoters of the lamprey Runx genes lack the tandem Runx-binding motifs that are highly conserved among the P1 promoters of gnathostome Runx1, Runx2 and Runx3 genes; instead these promoters contain dispersed single Runx-binding motifs. The 3'UTR of lamprey RunxB contains binding sites for miR-27 and miR-130b/301ab, which are conserved in mammalian Runx1 and Runx3, respectively. Overall, the Runx genes in lamprey seem to have experienced a different evolutionary trajectory from that of gnathostome Runx genes which are highly conserved all the way from cartilaginous fishes to mammals.

  18. Variable maturation and oviposition by female Schistosoma japonicum in mice: the effects of irradiation of the host prior to infection

    Energy Technology Data Exchange (ETDEWEB)

    Cheever, A.W.; Duvall, R.H.

    1987-11-01

    The maturation of female Schistosoma japonicum was found to vary greatly within each of two Philippine strains of this parasite and some females did not contain uterine eggs 7 to 15 weeks after infection while others contained numerous eggs before the fifth week of infection. It was found that female worms containing less than 20 uterine eggs contributed little to the accumulation of eggs in the tissues of infected mice. Such worms also generally appeared to be immature. The variable rate of maturation of worms is likely to have profound effects on the immune reactions of mice as well as on the pathologic response to infection. Systematic delay in oviposition was serendipitously found in worms from mice which had been irradiated for other purposes prior to exposure to S. japonicum, and from the fourth to the sixth week after infection egg production by worms in irradiated mice lagged well behind that in intact mice. Seven to 10 weeks after infection these worms were laying normal numbers of eggs, as judged by egg passage per worm pair in the feces and the accumulation of eggs in the tissues. S. mansoni developed normally in irradiated mice.

  19. Single- or mixed-sex Schistosoma japonicum infections of intermediate host snails in hilly areas of Anhui, China.

    Science.gov (United States)

    Shi, Hui-Ping; Lu, Da-Bing; Shen, Lei; Shi, Tan; Gu, Jian

    2014-02-01

    Schistosomiasis japonicum is one of the most serious communicable diseases, and the transmission of the parasite is dependent of its complex life cycle on which many factors can have an impact. Multiple infections comprising both male and female schistosome within snail intermediate hosts, for example, would facilitate parasite transmission. However, no research on Schistosoma japonicum communities in field-collected Oncomelania hupensis hupensis in relation to schistosome sex has been reported. Therefore, snail survey was performed in a hilly region of Anhui, China, and single- or mixed-sex schistosome infections of snails were detected with final host mouse infection. A total of 8,563 snails were sampled in the field, and 67 were identified with schistosome infections. Of these infected snails, 46 were selected for final host infection. From this, 21 snails were infected with female schistosome, 23 with males and 2 with both males and females. More worms were recovered for snails with mixed-sex infections than with single-sex infection and for snails with male schistosome infection than with female infection (Psnails was significantly higher than would be expected if randomly distributed (Psnails was nearly equal and up to 95.65 % (44/46) of infected snails were single-sex infection. Schistosome infections in snails collected from the hilly area of Anhui Province were not randomly distributed but over-dispersed.

  20. Expression profile of the Schistosoma japonicum degradome reveals differential protease expression patterns and potential anti-schistosomal intervention targets.

    Science.gov (United States)

    Liu, Shuai; Cai, Pengfei; Piao, Xianyu; Hou, Nan; Zhou, Xiaosu; Wu, Chuang; Wang, Heng; Chen, Qijun

    2014-10-01

    Blood fluke proteases play pivotal roles in the processes of invasion, nutrition acquisition, immune evasion, and other host-parasite interactions. Hundreds of genes encoding putative proteases have been identified in the recently published schistosome genomes. However, the expression profiles of these proteases in Schistosoma species have not yet been systematically analyzed. We retrieved and culled the redundant protease sequences of Schistosoma japonicum, Schistosoma mansoni, Echinococcus multilocularis, and Clonorchis sinensis from public databases utilizing bioinformatic approaches. The degradomes of the four parasitic organisms and Homo sapiens were then comparatively analyzed. A total of 262 S. japonicum protease sequences were obtained and the expression profiles generated using whole-genome microarray. Four main clusters of protease genes with different expression patterns were identified: proteases up-regulated in hepatic schistosomula and adult worms, egg-specific or predominantly expressed proteases, cercaria-specific or predominantly expressed proteases, and constantly expressed proteases. A subset of protease genes with different expression patterns were further validated using real-time quantitative PCR. The present study represents the most comprehensive analysis of a degradome in Schistosoma species to date. These results provide a firm foundation for future research on the specific function(s) of individual proteases and may help to refine anti-proteolytic strategies in blood flukes.

  1. Recombinant T2 RNase protein of Schistosoma japonicum inhibits expression of α-SMA in LX-2 cells.

    Science.gov (United States)

    Wang, Jianxin; Peng, Wenxia; Feng, Jinrong; Zhu, Dandan; Chen, Jinling; Sun, Xiaolei; Lyu, Lei; Ju, Shaoqing; Duan, Yinong

    2016-10-01

    Recombinant T2 RNase glycoprotein, which showed a certain degree of homology to Omega-1 from Schistosoma mansoni eggs, was expressed in adult worms of Schistosoma japonicum, but not in eggs of S. japonicum. The direct biological role of the recombinant T2 RNase protein in activation of hepatic stellate cells (HSCs) remains unknown. In the present study, the immortalized human HSC line (LX-2 cells) was treated with the recombinant T2 RNase protein at indicated concentrations for various time points in vitro. The expression levels of α-smooth muscle actin (α-SMA) and Smad4 were detected by Western blot. The results showed that the recombinant T2 RNase protein significantly diminished the expression levels of α-SMA and Smad4 in LX-2 cells. The upregulated expression levels of α-SMA and Smad4 by TGF-β1 in LX-2 cells were both suppressed by the recombinant T2 RNase protein. These data suggest that the recombinant T2 RNase protein may be a potential target of therapeutic strategy for the treatment of hepatic fibrosis.

  2. MicroRNAs Are Involved in the Regulation of Ovary Development in the Pathogenic Blood Fluke Schistosoma japonicum

    Science.gov (United States)

    Hu, Chao; Peng, Jinbiao; Luo, Rong; Zhou, Chunjing; Liu, Juntao; Lin, Jiaojiao; Jin, Youxin; Davis, Richard E.; Cheng, Guofeng

    2016-01-01

    Schistosomes, blood flukes, are an important global public health concern. Paired adult female schistosomes produce large numbers of eggs that are primarily responsible for the disease pathology and critical for dissemination. Consequently, understanding schistosome sexual maturation and egg production may open novel perspectives for intervening with these processes to prevent clinical symptoms and to interrupt the life-cycle of these blood-flukes. microRNAs (miRNAs) are key regulators of many biological processes including development, cell proliferation, metabolism, and signal transduction. Here, we report on the identification of Schistosoma japonicum miRNAs using small RNA deep sequencing in the key stages of male-female pairing, gametogenesis, and egg production. We identified 38 miRNAs, including 10 previously unknown miRNAs. Eighteen of the miRNAs were differentially expressed between male and female schistosomes and during different stages of sexual maturation. We identified 30 potential target genes for 16 of the S. japonicum miRNAs using antibody-based pull-down assays and bioinformatic analyses. We further validated some of these target genes using either in vitro luciferase assays or in vivo miRNA suppression experiments. Notably, suppression of the female enriched miRNAs bantam and miR-31 led to morphological alteration of ovaries in female schistosomes. These findings uncover key roles for specific miRNAs in schistosome sexual maturation and egg production. PMID:26871705

  3. [SDS-PAGE and EITB analysis of the protein components of different isolates of Schistosoma japonicum in China and Japan].

    Science.gov (United States)

    Qiu, L; Liu, S; Xue, H; Zhang, Y; Li, H; Hu, Y; He, Y

    1999-01-01

    To make a supplementary observation on the protein components prepared from adult S. japonicum of 4 different isolates from Zhejiang, Jiangxi, Taiwan Provinces of China and Japan in origin and to observe antigen reactivity of the above-mentioned isolates together with adult worms from Anhwi, Hubei, Sichuan and Yunnan isolates against heterologous anti-Oncomelania h. hupensis (collected from Guangxi, China and Japan) sera by EITB. SDS-PAGE and EITB. SDS-PAGE showed that by Coomassie blue staining, male S. japonicum from Jiangxi, Zhejiang, Taiwan Provinces and Japan isolales revealed 7-17 bands while female worms revealed 1-6 bands. The protein patterns of Taiwan and Zhejiang male worm were similar, but slight difference could be seen above 81 kDa. By silver staining, male worms of the 4 isolates revealed 10-23 bands while female worms revealed 1-19 bands. Male worms from Japan not only showed less bands but also differed in their pattern as compared to those of other Chinese isolates. Results of EITB revealed that each of the 4 isolates had slightly different pattern but all of the tested isolates had common antigens with their heterologous snail hosts i.e., Oncomelania h. hupensis from Guangxi, China and Japan.

  4. Multiple vaccinations with UV- attenuated cercariae in pig enhance protective immunity against Schistosoma japonicum infection as compared to single vaccination.

    Science.gov (United States)

    Lin, Dandan; Tian, Fang; Wu, Haiwei; Gao, Yanan; Wu, Jingjiao; Zhang, Donghui; Ji, Minjun; McManus, Donald P; Driguez, Patrick; Wu, Guanling

    2011-06-10

    Schistosomiasis japonica is a major public health problem in the endemic areas of China, the Philippines, and Indonesia. To date, a vaccine has not been developed against this disease but immunization with UV-attenuated cercariae can induce a high level of protective immunity in Landrace/Yorkshire/Duroc crossbred pigs. To compare the efficacy of a single vaccination and multiple vaccinations with UV-attenuated Schistosoma japonicum cercariae, two groups of pigs received either one or three exposures to 10,000 cercariae attenuated with 400 μw UV. Pigs with a single immunization had a 59.33% reduction in adult worm burden, a 89.87% reduction in hepatic eggs and a 86.27% reduction in fecal eggs at eight weeks post-challenge (P vaccinated groups were higher than in the infection-control group. Triple vaccinations resulted in higher levels of antibodies, especially IgG2, compared with a single vaccination and IFN-γ levels increased with repeated immunization with UV-irradiated cercariae. The high levels of protection against S. japonicum infection can be achieved with a UV-attenuated vaccine in pigs, and that three vaccinations were possibly more effective than a single vaccination. Moreover, triple vaccinations evoked a more vigorous IFN-γ response and a stronger antibody-mediated response, especially an increase in the levels of IgG2 antibodies.

  5. Culturable bacteria in hydroponic cultures of moss Racomitrium japonicum and their potential as biofertilizers for moss production.

    Science.gov (United States)

    Tani, Akio; Akita, Motomu; Murase, Haruhiko; Kimbara, Kazuhide

    2011-07-01

    The use of Racomitrium japonicum, a drought resistant bryophyte used for roof-greening, is gradually increasing. However, its utilization is hampered by slow growth rate. Here we isolated culturable bacteria from hydroponic cultivation samples to identify isolates that could promote moss growth. Most of the isolates belonged to Pseudomonas, Rhodococcus, and Duganella species. The isolates were biochemically characterized according to their type of interaction with plants, i.e., production of auxin, siderophores, or hydrogen cyanate, growth in the absence of an added nitrogen source, calcium phosphate solubilization, utilization of sugars, polymers, or aliphatic compounds, and antifungal activity. The isolates were applied to sterile protonemata and non-sterile adult gametophytes of R. japonicum to evaluate their effect on plant growth. Furthermore, we isolated fungi that inhibited moss growth. Our results suggest that the microbial community structure in hydroponic cultures is important to stabilize moss production and the isolates that promote moss growth have potential to be utilized as biofertilizers for moss production. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  6. Gonadal Morphology and Gametogenesis in Japanese Red Coral Corallium japonicum (Octocorallia: Alcyonacea) Collected off Cape Ashizuri, Japan.

    Science.gov (United States)

    Sekida, Satoko; Iwasaki, Nozomu; Okuda, Kazuo

    2016-06-01

    Colonies of the Japanese red coral Corallium japonicum Kishinouye, 1903 collected off Cape Ashizuri, Japan were gonochoric and produced gonads in siphonozooids annually, mainly during the spring season. Polyp anatomy, gonadal morphology and gametogenesis in this species were revealed by light and electron microscopy. A siphonozooid had a pharynx with a prominent siphonoglyph and eight mesenteries: two sulcal, two asulcal, and four lateral. A rudimentary retractor was found on one side of each mesoglea of these mesenteries. The retractor arrangement in the siphonozooid was reverse of what was described in the autozooids of octocorals. Gonads initiated as small protrusions on the mesenteries, except in the asulcal ones, and even at an incipient stage they were covered with a sac-shaped thin layer of mesoglea, which was continuous with the mesoglea of mesenteries. Gastrodermis enveloped the complete outer surface of the thin layer of mesoglea throughout gametogenesis in both oocytes and sperm cysts. Oocytes produced many microvilli on their cortical surfaces beneath the thin layer of mesoglea concomitantly with the accumulation of lipid globules in the cells, whereas in sperm cysts spermatocytes and spermatids increased in number without microvilli production, followed by synchronous spermiogenesis involving remarkable changes in the shape and position of organelles. Based on the comparison of patterns in gonadal development between octocorals including C. japonicum, hexacorals and scyphozoans, octocoral and stauromedusa species may be characterized by the fact that gametogenesis never occurs in the matrix of mesoglea, but rather exclusively within the thin sac of mesoglea surrounded by gastrodermis.

  7. MicroRNAs Are Involved in the Regulation of Ovary Development in the Pathogenic Blood Fluke Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Lihui Zhu

    2016-02-01

    Full Text Available Schistosomes, blood flukes, are an important global public health concern. Paired adult female schistosomes produce large numbers of eggs that are primarily responsible for the disease pathology and critical for dissemination. Consequently, understanding schistosome sexual maturation and egg production may open novel perspectives for intervening with these processes to prevent clinical symptoms and to interrupt the life-cycle of these blood-flukes. microRNAs (miRNAs are key regulators of many biological processes including development, cell proliferation, metabolism, and signal transduction. Here, we report on the identification of Schistosoma japonicum miRNAs using small RNA deep sequencing in the key stages of male-female pairing, gametogenesis, and egg production. We identified 38 miRNAs, including 10 previously unknown miRNAs. Eighteen of the miRNAs were differentially expressed between male and female schistosomes and during different stages of sexual maturation. We identified 30 potential target genes for 16 of the S. japonicum miRNAs using antibody-based pull-down assays and bioinformatic analyses. We further validated some of these target genes using either in vitro luciferase assays or in vivo miRNA suppression experiments. Notably, suppression of the female enriched miRNAs bantam and miR-31 led to morphological alteration of ovaries in female schistosomes. These findings uncover key roles for specific miRNAs in schistosome sexual maturation and egg production.

  8. A cluster-randomised intervention trial against Schistosoma japonicum in the Peoples' Republic of China: bovine and human transmission.

    Directory of Open Access Journals (Sweden)

    Darren J Gray

    2009-06-01

    Full Text Available Zoonotic schistosomiasis japonica is a major public health problem in China. Bovines, particularly water buffaloes, are thought to play a major role in the transmission of schistosomiasis to humans in China. Preliminary results (1998-2003 of a praziquantel (PZQ-based pilot intervention study we undertook provided proof of principle that water buffaloes are major reservoir hosts for S. japonicum in the Poyang Lake region, Jiangxi Province.Here we present the results of a cluster-randomised intervention trial (2004-2007 undertaken in Hunan and Jiangxi Provinces, with increased power and more general applicability to the lake and marshlands regions of southern China. The trial involved four matched pairs of villages with one village within each pair randomly selected as a control (human PZQ treatment only, leaving the other as the intervention (human and bovine PZQ treatment. A sentinel cohort of people to be monitored for new infections for the duration of the study was selected from each village. Results showed that combined human and bovine chemotherapy with PZQ had a greater effect on human incidence than human PZQ treatment alone.The results from this study, supported by previous experimental evidence, confirms that bovines are the major reservoir host of human schistosomiasis in the lake and marshland regions of southern China, and reinforce the rationale for the development and deployment of a transmission blocking anti-S. japonicum vaccine targeting bovines.Australian New Zealand Clinical Trials Registry ACTRN12609000263291.

  9. Upregulated Expression of Cytotoxicity-Related Genes in IFN-γ Knockout Mice with Schistosoma japonicum Infection

    Directory of Open Access Journals (Sweden)

    Xiaotang Du

    2011-01-01

    Full Text Available It is well accepted that IFN-γ is important to the development of acquired resistance against murine schistosomiasis. However, the in vivo role of this immunoregulatory cytokine in helminth infection needs to be further investigated. In this study, parasite burden and host immune response were observed in IFN-γ knockout mice (IFNg KO infected with Schistosoma japonicum for 6 weeks. The results suggested that deficiency in IFN-γ led to decreased egg burden in mice, with low schistosome-specific IgG antibody response and enhanced activation of T cells during acute infection. Microarray and qRT-PCR data analyses showed significant upregulation of some cytotoxicity-related genes, including those from the granzyme family, tumor necrosis factor, Fas Ligand, and chemokines, in the spleen cells of IFNg KO mice. Furthermore, CD8+ cells instead of NK cells of IFNg KO mice exhibited increased transcription of cytotoxic genes compared with WT mice. Additionally, Schistosoma japonicum-specific egg antigen immunization also could activate CD8+ T cells to upregulate the expression of cytotoxic genes in IFNg KO mice. Our data suggest that IFN-γ is not always a positive regulator of immune responses. In certain situations, the disruption of IFN-γ signaling may up-regulate the cytotoxic T-cell-mediated immune responses to the parasite.

  10. Compatibilidade do tratamento de sementes de feijão-caupi com fungicidas e inoculação com estirpes de Bradyrhizobium

    Directory of Open Access Journals (Sweden)

    Manoel Luiz da Silva Neto

    2013-01-01

    Full Text Available O objetivo deste trabalho foi avaliar a compatibilidade do tratamento de sementes com fungicidas e a inoculação com estirpes de Bradyrhizobium em feijão-caupi. Em laboratório, avaliou-se a sobrevivência de células nas sementes da cultivar BRS Guariba, tratadas ou não com fungicidas (fludioxonil, carbendazim, carbendazim + thiram e carboxin + thiram e inoculadas ou não com Bradyrhizobium (estirpes UFLA3-84, BR 3267, INPA3-11B e BR 3262. Em casa de vegetação, conduziu-se experimento em vasos de Leonard, com os mesmos tratamentos. Foram avaliados: massa de matéria seca da parte aérea, além de número e massa de nódulos 25 dias após a emergência das plantas. No campo, dois experimentos foram conduzidos, tendo-se utilizado a estirpe BR 3262, com aplicação de fungicidas nas sementes: um em área de primeiro cultivo e outro em área cultivada anteriormente com culturas anuais. Avaliaram-se, aos 35 dias, o número de nódulos, a massa de nódulos secos e a massa de matéria seca da parte aérea, e, na colheita, a produtividade de grãos. Os fungicidas não tiveram efeito significativo sobre a sobrevivência de Bradyrhizobium, a nodulação das plantas e o rendimento de grãos, que, em média, foi superior a 1.200 kg ha-1. O tratamento de sementes de feijão-caupi com fungicidas é compatível com a inoculação das estirpes avaliadas.

  11. Combined IL-12 Plasmid and Recombinant SjGST Enhance the Protective and Anti-pathology Effect of SjGST DNA Vaccine Against Schistosoma japonicum

    National Research Council Canada - National Science Library

    Cheng, Po-Ching; Lin, Ching-Nan; Peng, Shih-Yi; Kang, Tsung-Fu; Lee, Kin-Mu

    2016-01-01

    ...) has previously been reported to achieve a worm reduction rate of 42-44%. To improve the efficiency of the vaccine against Schistosoma japonicum, we immunized mice with a combination of pcDNA vector-encoded 26-kDa SjGST (pcDNA/SjGST...

  12. Tolerância de estirpes e isolados de Bradyrhizobium e de Azorhizobium a zinco, cádmio e cobre "in vitro"

    Directory of Open Access Journals (Sweden)

    I. C. B. Trannin

    2001-06-01

    Full Text Available Com o objetivo de avaliar a tolerância a metais pesados "in vitro" de estirpes inoculantes (I, isolados de solos contaminados com metais (ISC e de solos não contaminados (ISNC de Bradyrhizobium, simbiontes de Enterolobium contortisiliquum (tamboril e de Acacia mangium (acácia e de Azorhizobium, simbiontes de Sesbania virgata (sesbânia, foram realizados dois experimentos. No primeiro, dez estirpes e, ou, isolados para cada espécie vegetal foram testados em meio YMA modificado por adição de tampões biológicos (HEPES e MES, suplementado com diferentes concentrações de Cu, Cd e Zn. Cobre e Cd foram testados em concentrações de 0 a 40 mg L-1, para ambos os gêneros, enquanto Zn variou de 0 a 1.000 mg L-1, para Bradyrhizobium, e de 0 a 500 mg L-1, para Azorhizobium. O crescimento de rizóbio nas diferentes concentrações de metais foi avaliado com atribuição de valores para os padrões observados (0 a 5. Os ISC de ambos os gêneros foram mais tolerantes, mas Bradyrhizobium tolerou Zn (800 mg L-1 até duas vezes e Cu (40 mg L-1 até oito vezes mais que Azorhizobium. No segundo experimento, estirpes e isolados tolerantes (T, sensíveis (S e de tolerância média (TM a metais selecionados em meio YMA modificado foram estudados em soluções aquosas com diferentes concentrações de Cu (0 a 0,01 mg L-1, Cd e Zn (0 a 1,0 mg L-1. A avaliação do número de células viáveis em soluções de metais foi feita por contagem das unidades formadoras de colônia em 0, 24, 48, 72 e 96 h de incubação, pelo método das diluições sucessivas e inoculação em YMA. Embora as soluções de metais tenham sido mais discriminatórias quanto a tolerância a metais que o meio YMA, estes dois métodos mostraram que: (a Azorhizobium foi mais sensível que Bradyrhizobium, (b os ISC de ambos os gêneros foram mais tolerantes do que os ISNC e (c a ordem de toxidez dos metais foi Cu > Cd > Zn.

  13. INFLUENCIA DE LA CONCENTRACIÓN DE MELAZA Y EXTRACTO ACUOSO DE SOYA SOBRE LA VELOCIDAD ESPECÍFICA DE CRECIMIENTO DE Bradyrhizobium elkanii ICA 8001

    OpenAIRE

    Gretel Gómez; María C. Nápoles; R. Núñez; Martínez, J.

    2008-01-01

    La cepa ICA 8001 de Bradyrhizobium elkanii es ampliamente usada en Cuba para la producción de inoculantes para soya. Uno de los medios de cultivos diseñados para la multiplicación de este microorganismo contiene melaza y extracto acuoso de soya, elementos de variable composición y origen, que aportan cantidades notables de sustratos carbonados. Por esta causa, se determinó la concentración de carbohidratos totales solubles y carbohidratos reductores, así como los contenidos de N, P, K, Ca, Mg...

  14. Development of a Streptococcus gordonii vaccine strain expressing Schistosoma japonicum Sj-F1 and evaluation of using this strain for intranasal immunization in mice.

    Science.gov (United States)

    Wang, Linqian; Liu, Wei; Yang, Ming; Peng, Dan; Chen, Liyu

    2013-04-01

    Schistosomiasis is a worldwide parasitic disease. Currently, chemotherapy is the main effective method to treat schistosomiasis; however, it does not prevent reinfection. No effective vaccine is currently available to prevent schistosomiasis. Sj-F1 (GenBank accession number AY261995) is a novel gene that was discovered through screening adult Schistosoma japonicum worm cDNA library with female S. japonicum antigen-immunized sera. Streptococcus gordonii, a normal inhabitant of the human oral cavity, has been a prime candidate in recent investigations toward developing a live oral vaccine vector. One of the approaches for the surface expression of heterologous antigens in S. gordonii is to surface-localize them with the M6 protein from Streptococcus pyogenes. Here, we develop a recombinant S. gordonii strain that expresses the M6-Sj-F1 fusion protein on the bacterial surface. Intranasal immunization in mice with such M6-Sj-F1-expressing S. gordonii bacteria induced strong serum IgG, serum IgA, and saliva IgA against Sj-F1. The results of protective immunity against a challenge with cercariae of S. japonicum showed statistically significant protection following this treatment, with a worm reduction rate of 21.45% and an egg reduction rate of 34.77%. Our data indicate that the described M6-Sj-F1-expressing S. gordonii is highly immunogenic and can partially protect mice from challenge infection with S. japonicum. Intranasal immunization with recombinant S. gordonii may be an alternative to developing a novel S. japonicum vaccine in a safe, effective, and feasible way.

  15. Therapeutic effect of Schistosoma japonicum cystatin on bacterial sepsis in mice.

    Science.gov (United States)

    Li, Huihui; Wang, Shushu; Zhan, Bin; He, Wenxin; Chu, Liang; Qiu, Dapeng; Li, Nan; Wan, Yongkun; Zhang, Hui; Chen, Xingzhi; Fang, Qiang; Shen, Jilong; Yang, Xiaodi

    2017-05-08

    Sepsis is a life-threatening complication of an infection and remains one of the leading causes of mortality in surgical patients. Bacteremia induces excessive inflammatory responses that result in multiple organ damage. Chronic helminth infection and helminth-derived materials have been found to immunomodulate host immune system to reduce inflammation against some allergic or inflammatory diseases. Schistosoma japonicum cystatin (Sj-Cys) is a cysteine protease inhibitor that induces regulatory T-cells and a potential immunomodulatory. The effect of Sj-Cys on reducing sepsis inflammation and mortality was investigated. Sepsis was induced in BALB/c mice using cecal ligation and puncture (CLP), followed by intraperitoneal injection of different doses (10, 25 or 50 μg) of recombinant Sj-Cys (rSj-Cys). The therapeutic effect of rSj-Cys on sepsis was evaluated by observing the survival rates of mice for 96 h after CLP and the pathological injury of liver, kidney and lung by measuring the levels of alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN) and creatinine (Cr) in sera and the tissue sections pathology, and the expression of MyD88 in liver, kidney and lung tissues. The immunological mechanism was investigated by examining pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and IL-10 and TGF-β1 in mice sera and in culture of macrophages stimulated by lipopolysaccharides (LPS). rSj-Cys treatment provided significant therapeutic effects on CLP-induced sepsis in mice demonstrated with increased survival rates, alleviated overall disease severity and tissue injury of liver, kidney and lung. The rSj-Cys conferred therapeutic efficacy was associated with upregualted IL-10 and TGF-β1 cytokines and reduced pro-inflammatory cytokines TNF-α, IL-6, IL-1β. MyD88 expression in liver, kidney and lung tissues of rSj-Cys-treated mice was reduced. In vitro assay with macrophages also showed that rSj-Cys inhibited the release of pro

  16. The anterior esophageal region of Schistosoma japonicum is a secretory organ.

    Science.gov (United States)

    Li, Xiao Hong; Stark, Meg; Vance, Gillian M; Cao, Jian Ping; Wilson, R Alan

    2014-12-10

    The esophagus of blood-feeding schistosomes has been largely neglected although its posterior portion was designated as a gland decades ago. However, we recently showed it plays a pivotal role in blood processing. It is clearly demarcated into anterior and posterior compartments, both surrounded by a mass of cell bodies. Feeding movies revealed that erythrocytes accumulate in the anterior compartment before entering the posterior, indicating that a distinct process is executed there. We therefore investigated ultrastructural aspects and possible functions of the anterior region. The heads of adult Schistosoma japonicum were detached and prepared for both transmission and scanning electron microscopy to define the detailed ultrastructure of the anterior esophagus. Cryosections of heads were also prepared for immunocytochemistry and confocal microscopy to define the pattern of intrinsic host antibody binding in the anterior esophageal lining. The anterior syncytial lining of the esophagus is highly extended by long, thin corrugations of cytoplasm projecting towards the lumen. Strikingly in the male worm, the tips of the corrugations are further expanded by numerous threads of cytoplasm, producing a spaghetti-like appearance in the central lumen. Flattened, pitted cytoplasmic plates are interspersed in the tangled mass of threads. Abundant, morphologically distinct light vesicles of varied size and contents are manufactured in the cell bodies, from where they traffic through cytoplasmic connections to the corrugations and out to the tips. Clusters of vesicles accumulate in expanded tips in males, together with occasional mitochondria whilst females have more mitochondria but fewer vesicles. The membranous contents of light vesicles are secreted mainly from the tips, but also from the sides of the corrugations. They coat the surfaces and then form organised self-adherent membrane figures when shed into the lumen. Host antibody binds strongly in a characteristic pattern

  17. Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of Schistosoma japonicum.

    Science.gov (United States)

    Sun, Kui; Xing, Weiwei; Yu, Xinling; Fu, Wenliang; Wang, Yuanyuan; Zou, Minji; Luo, Zhihong; Xu, Donggang

    2016-08-31

    With the continuous decline in prevalence and intensity of Schistosoma japonicum infection in China, more accurate and sensitive methods suitable for field detection become much needed for schistosomiasis control. Here, a novel rapid and visual detection method based on the combination of recombinase polymerase amplification (RPA) and lateral flow dipstick (LFD) was developed to detect S. japonicum DNA in fecal samples. The LFD-RPA assay targeting SjR2 could detect 5 fg S. japonicum DNA, which was identical to qPCR and real-time RPA assay, and showed no cross-reaction with other parasites. The detection could be finished within 15-20 min at a wide temperature range (25-45 °C), and the results could be visualized by naked eye. The diagnostic validity of LFD-RPA assay was further assessed with 14 fecal samples of infected patients diagnosed by Kato-Katz method and 31 fecal samples of healthy persons, and compared with that of Enzyme-linked immunosorbent assay (ELSIA) and Indirect Hemagglutination Assay (IHA). The LFD-RPA assay showed 92.68 % sensitivity, 100 % specificity and excellent diagnostic agreement with the gold standard Kato-Katz test (k = 0.947, Z = 6.36, P < 0.001), whereas ELISA showed 85.71 % sensitivity, 93.55 % specificity, and substantial diagnostic agreement (k = 0.793, Z = 5.31, P < 0.001), and IHA showed 78.57 % sensitivity, 83.87 % specificity, and moderate diagnostic agreement (k = 0.600, Z = 4.05, P < 0.001), indicating that the LFD-RPA was much better than the traditional methods. The LFD-RPA assay established by us is a sensitive, specific, rapid and convenient method for the diagnosis of schistosomiasis, and shows a great potency in field application.

  18. A Proteomic Approach of Bradyrhizobium/Aeschynomene Root and Stem Symbioses Reveals the Importance of the fixA Locus for Symbiosis

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    Nathanael Delmotte

    2014-02-01

    Full Text Available Rhizobia are soil bacteria that are able to form symbiosis with plant hosts of the legume family. These associations result in the formation of organs, called nodules in which bacteria fix atmospheric nitrogen to the benefit of the plant. Most of our knowledge on the metabolism and the physiology of the bacteria during symbiosis derives from studying roots nodules of terrestrial plants. Here we used a proteomics approach to investigate the bacterial physiology of photosynthetic Bradyrhizobium sp. ORS278 during the symbiotic process with the semi aquatical plant Aeschynomene indica that forms root and stem nodules. We analyzed the proteomes of bacteria extracted from each type of nodule. First, we analyzed the bacteroid proteome at two different time points and found only minor variation between the bacterial proteomes of 2-week- and 3-week-old nodules. High conservation of the bacteroid proteome was also found when comparing stem nodules and root nodules. Among the stem nodule specific proteins were those related to the phototrophic ability of Bradyrhizobium sp. ORS278. Furthermore, we compared our data with those obtained during an extensive genetic screen previously published. The symbiotic role of four candidate genes which corresponding proteins were found massively produced in the nodules but not identified during this screening was examined. Mutant analysis suggested that in addition to the EtfAB system, the fixA locus is required for symbiotic efficiency.

  19. Induced systemic resistance and symbiotic performance of peanut plants challenged with fungal pathogens and co-inoculated with the biocontrol agent Bacillus sp. CHEP5 and Bradyrhizobium sp. SEMIA6144.

    Science.gov (United States)

    Figueredo, María Soledad; Tonelli, María Laura; Ibáñez, Fernando; Morla, Federico; Cerioni, Guillermo; Del Carmen Tordable, María; Fabra, Adriana

    2017-04-01

    Synergism between beneficial rhizobacteria and fungal pathogens is poorly understood. Therefore, evaluation of co-inoculation of bacteria that promote plant growth by different mechanisms in pathogen challenged plants would contribute to increase the knowledge about how plants manage interactions with different microorganisms. The goals of this work were a) to elucidate, in greenhouse experiments, the effect of co-inoculation of peanut with Bradyrhizobium sp. SEMIA6144 and the biocontrol agent Bacillus sp. CHEP5 on growth and symbiotic performance of Sclerotium rolfsii challenged plants, and b) to evaluate field performance of these bacteria in co-inoculated peanut plants. The capacity of Bacillus sp. CHEP5 to induce systemic resistance against S. rolfsii was not affected by the inoculation of Bradyrhizobium sp. SEMIA6144. This microsymbiont, protected peanut plants from the S. rolfsii detrimental effect, reducing the stem wilt incidence. However, disease incidence in plants inoculated with the isogenic mutant Bradyrhizobium sp. SEMIA6144 V2 (unable to produce Nod factors) was as high as in pathogen challenged plants. Therefore, Bradyrhizobium sp. SEMIA6144 Nod factors play a role in the systemic resistance against S. rolfsii. Bacillus sp. CHEP5 enhanced Bradyrhizobium sp. SEMIA6144 root surface colonization and improved its symbiotic behavior, even in S. rolfsii challenged plants. Results of field trials confirmed the Bacillus sp. CHEP5 ability to protect against fungal pathogens and to improve the yield of extra-large peanut seeds from 2.15% (in Río Cuarto) to 16.69% (in Las Vertientes), indicating that co-inoculation of beneficial rhizobacteria could be a useful strategy for the peanut production under sustainable agriculture system. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. DISTRIBUSI HABITAT Oncomelania hupensis lindoensis, KEONG PERANTARA Schistosoma japonicum DI DATARAN TINGGI LINDU, KABUPATEN SIGI, SULAWESI TENGAH

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    Triwibowo Ambar Garjito

    2015-01-01

    Full Text Available Abstract Oncomelania hupensis lindoensissnail and its habitat has an important role in the transmission of schistosomiasis in Central Sulawesi, particularly in three isolated areas, Lindu valley, Napu valley and Bada valley. In a part of Schistosomiasis life cycle, inside the snail, Schistosoma japonicummiracidia will undergo a series of stages as sporocyst and cercaria. People are infected by cercaria, the infective stage of S. japonicum.This study were conducted to reconfirm the distribution of O. h. lindoensishabitats in Lindu valley area. The snails were searched and collected in the suspected habitat using ring-sample and man per minute methods by skilled staffs from VBDRU Donggala and Schistosomasis laboratory plus trained local people in the collections. Data on the distribution of snail habitats were recorded by using GPS. Snails and vegetation in the habitats were collected for further analysis in the laboratory. A total of 129 snail habitat were recorded in Lindu valley, consisting of 135 old foci and 1 new focus. In this area, a total of 61 foci are still active of snail habitats. Foci are distributed in several types of habitat, i.e. abandon rice fields, ditches, springs, dry farming, shrubs and forest. Each type habitat has a relative similar vegetation species. The infection rates of O. h. lindoensiswith cercariae in Anca, Tomado dan Puroo villages were 5.27%, 3.19% and 7.58% respectively. These results indicate that the Schistosomiasis transmission is still going on in Lindu valley.Keywords : Distribution, Oncomelania hupensis lindoensis, Habitat, Schistosomiasis, Lindu Valley, Sulawesi TengahAbstrakKeberadaan keong Oncomelania hupensis lindoensis dan habitatnya mempunyai peranan penting terhadap terjadinya penularan Skistosomiasis di Sulawesi Tengah, khususnya di 3 daerah endemis yang cukup terisolasi, yaitu Dataran tinggi Lindu, Dataran Tinggi Napu dan Dataran Tinggi Bada. Di dalam keong tersebut, mirasidium Schistosoma

  1. Possible effects of the Three Gorges dam on the transmission of Schistosoma japonicum on the Jiang Han plain, China.

    Science.gov (United States)

    Xu, X J; Wei, F H; Yang, X X; Dai, Y H; Yu, G Y; Chen, L Y; Su, Z M

    2000-06-01

    The Three Gorges dam, under construction on the Yangtze River in China, might affect the transmission of Schistosoma japonicum on the Jiang Han plain, which is downstream of the dam. To study this possibility, the prevalence of schistosomiasis was investigated in relation to a range of malacological, hydrological and meteorological factors. The general water level in the Yangzte over a year had a marked effect on the distribution of the intermediate host (Oncomelania hupensis) and the prevalence of human schistosomiasis in that year. Disease prevalence showed significant correlations with the density of the snail hosts, the level of the water table, annual rainfall, yearly evaporation, and altitude. Once the dam is complete, the flow of water downstream will probably be maintained at a level between those currently occurring in flood and dry weather, and this may have implications for schistosome transmission. Systematic monitoring is necessary to investigate the impact of the environmental changes brought about by the dam on transmission.

  2. Genetic variability among Schistosoma japonicum isolates from the Philippines, Japan and China revealed by sequence analysis of three mitochondrial genes.

    Science.gov (United States)

    Chen, Fen; Li, Juan; Sugiyama, Hiromu; Zhou, Dong-Hui; Song, Hui-Qun; Zhao, Guang-Hui; Zhu, Xing-Quan

    2015-02-01

    The present study examined sequence variability in the mitochondrial (mt) protein-coding genes cytochrome b (cytb), NADH dehydrogenase subunits 2 and 6 (nad2 and nad6) among 24 isolates of Schistosoma japonicum from different endemic regions in the Philippines, Japan and China. The complete cytb, nad2 and nad6 genes were amplified and sequenced separately from individual schistosome. Sequence variations for isolates from the Philippines were 0-0.5% for cytb, 0-0.6% for nad2, and 0-0.9% for nad6. Variation was 0-0.5%, 0.1-0.8%, 0-0.7% for corresponding genes for schistosome samples from mainland China. For worms in Japan, genetic variations were 0-0.2%, 0.1-0.2% and 0 for the three genes, respectively. Sequence variations were 0-1.0%, 0-1.8% and 0-1.1% for cytb, nad2 and nad6, respectively, among schistosome isolates from different geographical strains in the Philippines, Japan and China. Of the three countries, lowest sequence variations were found between isolates from mainland China and the Philippines and highest were detected between Japan and the Philippines in three mtDNA genes. Phylogenetic analyses based on the combined sequences of cytb, nad2 and nad6 revealed that all isolates in the Philippines clustered together sistered to samples from Yunnan and Zhejiang provinces in China, while isolates from Yamanashi in Japan were in a solitary clade. These results demonstrated the usefulness of the combined three mtDNA sequences for studying genetic diversity and population structure among S. japonicum isolates from the Philippines, China and Japan.

  3. The nature and combination of subunits used in epitope-based Schistosoma japonicum vaccine formulations affect their efficacy

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    Liu Feng

    2010-11-01

    Full Text Available Abstract Background Schistosomiasis remains a major public health problem in endemic countries and is caused by infections with any one of three primary schistosome species. Although there are no vaccines available to date, this strategy appears feasible since natural immunity develops in individuals suffering from repeated infection during a lifetime. Since vaccinations resulting in both Th1- and Th2-type responses have been shown to contribute to protective immunity, a vaccine formulation with the capacity for stimulating multiple arms of the immune response will likely be the most effective. Previously we developed partially protective, single Th- and B cell-epitope-based peptide-DNA dual vaccines (PDDV (T3-PDDV and B3-PDDV, respectively capable of eliciting immune responses against the Schistosoma japonicum 22.6 kDa tegument antigen (Sj22.6 and a 62 kDa fragment of myosin (Sj62, respectively. Results In this study, we developed PDDV cocktails containing multiple epitopes of S. japonicum from Sj22.6, Sj62 and Sj97 antigens by predicting cytotoxic, helper, and B-cell epitopes, and evaluated vaccine potential in vivo. Results showed that mice immunized with a single-epitope PDDV elicited either Tc, Th, or B cell responses, respectively, and mice immunized with either the T3- or B3- single-epitope PDDV formulation were partially protected against infection. However, mice immunized with a multicomponent (3 PDDV components formulation elicited variable immune responses that were less immunoprotective than single-epitope PDDV formulations. Conclusions Our data show that combining these different antigens did not result in a more effective vaccine formulation when compared to each component administered individually, and further suggest that immune interference resulting from immunizations with antigenically distinct vaccine targets may be an important consideration in the development of multicomponent vaccine preparations.

  4. Utilization of ELISA using thioredoxin peroxidase-1 and tandem repeat proteins for diagnosis of Schistosoma japonicum infection among water buffaloes.

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    Jose Ma M Angeles

    Full Text Available BACKGROUND: The presence of animal reservoirs in Schistosoma japonicum infection has been a major obstacle in the control of schistosomiasis. Previous studies have proven that the inclusion of control measures on animal reservoir hosts for schistosomiasis contributed to the decrease of human cases. Animal surveillance should therefore be included to strengthen and improve the capabilities of current serological tests. METHODOLOGY/PRINCIPAL FINDINGS: Thioredoxin peroxidase-1 (SjTPx-1 and four tandem repeat proteins (Sj1TR, Sj2TR, Sj4TR, Sj7TR were initially evaluated against human sera. The previous test showed high sensitivity and specificity for antibody detection against SjTPx-1 and Sj7TR. In this study, the immunodiagnostic potential of these recombinant proteins was evaluated using enzyme-linked immunoassay on 50 water buffalo serum samples collected in Cagayan, the Philippines as compared with the soluble egg antigen (SEA. For specificity, 3 goat serum samples positive with Fasciola hepatica were used and among the antigens used, only SEA showed cross-reaction. Stool PCR targeting the S. japonicum 82 bp mitochondrial NAD 1 gene was done to confirm the true positives and served as the standard test. Twenty three samples were positive for stool PCR. SjTPx-1 and Sj1TR gave the highest sensitivity among the recombinant proteins tested for water buffalo samples with 82.61% and 78.26% respectively which were higher than that of SEA (69.57%. CONCLUSIONS/SIGNIFICANCE: These results prove that SjTPx-1 works both for humans and water buffaloes making it a good candidate antigen for zoonotic diagnosis. Sj1TR showed good results for water buffaloes and therefore can also be used as a possible candidate for detecting animal schistosome infection.

  5. Exosomes Derived from Dendritic Cells Treated with Schistosoma japonicum Soluble Egg Antigen Attenuate DSS-Induced Colitis

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    Lifu Wang

    2017-09-01

    Full Text Available Exosomes are 30–150 nm small membrane vesicles that are released into the extracellular medium via cells that function as a mode of intercellular communication. Dendritic cell (DC-derived exosomes modulate immune responses and prevent the development of autoimmune diseases. Moreover, Schistosoma japonicum eggs show modulatory effects in a mouse model of colitis. Therefore, we hypothesized that exosomes derived from DCs treated with S. japonicum soluble eggs antigen (SEA; SEA-treated DC exosomes would be useful for treating inflammatory bowel disease (IBD. Exosomes were purified from the supernatant of DCs treated or untreated with SEA and identified via transmission electron microscopy, western blotting and NanoSight. Acute colitis was induced via the administration of dextran sulfate sodium (DSS in drinking water (5.0%, wt/vol. Treatment with exosomes was conducted via intraperitoneal injection (i.p.; 50 μg per mouse from day 0 to day 6. Clinical scores were calculated based on weight loss, stool type, and bleeding. Colon length was measured as an indirect marker of inflammation, and colon macroscopic characteristics were determined. Body weight loss and the disease activity index of DSS-induced colitis mice decreased significantly following treatment with SEA-treated DC exosomes. Moreover, the colon lengths of SEA-treated DC exosomes treated colitis mice improved, and their mean colon macroscopic scores decreased. In addition, histologic examinations and histological scores showed that SEA-treated DC exosomes prevented colon damage in acute DSS-induced colitis mice. These results indicate that SEA-treated DC exosomes attenuate the severity of acute DSS-induced colitis mice more effectively than DC exosomes. The current work suggests that SEA-treated DC exosomes may be useful as a new approach to treat IBD.

  6. A multivariate analysis of the relationship between work ability and S. japonicum infection in Dongting Lake Region, in China Análise multivariada da relação entre capacidade de trabalho e infecção por S. japonicum na região dos lagos de Dongting, China

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    Li Yuesheng

    1993-08-01

    Full Text Available A cross-sectional case-control study on the association between the reduced work ability and S. japonicum infection was carried out in a moderate endemic area for schistosomiasis japonica in the southern part of Dongting lake in China. A total of 120 cases with reduced work ability and 240 controls paired to the case by age, sex, occupation and without reduced work ability, participated in the study. The mean age for individuals was 37.6 years old (21-60, the ratio of male: female was 60:40, the prevalence of S. japonicum in the individuals was 28.3%. The results obtained in this study showed that the infection of S. japonicum in case and control groups was 49.2% (59/120 and 17.9% (43/240, respectively. Odds ratio for reduced work ability among those who had schistosomiasis was 4.34 (95%, confidence interval was 2.58-7.34, and among those who had S. japonicum infection (egg per gram > 100 was up to 12.67 (95%, confidence interval was 3.64-46.39. After odds ratio was adjusted by multiple logistic regression, it was confirmed that heavier intensity of S. japonicum infection and splenomegaly due to S. japonicum infection were the main risk factors for reduced work ability in the population studied.Um estudo seccional de casos controles da associação entre a capacidade reduzida para o trabalho e a infecção por S. japonicum foi levada a efeito em região moderadamente endêmica para esquistossomose japônica na parte sul do lago Dongting, China. Um total de 120 casos com redução da capacidade de trabalho e 240 controles pareados no que diz respeito a idade, sexo, ocupação sem redução da capacidade de trabalho. A idade média dos pacientes foi 37,6 anos (21-60 e a relação masculino:feminino foi 60:40. A prevalência do S. japonicum foi de 28,3%. Os resultados obtidos neste estudo mostraram que a infecção nos casos e no grupo controle foi 49,2% (59/120 e 17,9% (43/240 respectivamente. A média para redução da capacidade de trabalho

  7. [Comparison of collagen fiber staining between Van-Gieson staining and Masson trichrome staining of hepatic specimens in mice with Schistosoma japonicum infection].

    Science.gov (United States)

    Huang, Da-Ke; Zhang, Yu-Xia; Man, Su-Qin; Yu, Fa-Zhi; Shen, Ji-Jia

    2012-08-01

    To compare the effects of collagen fiber staining between Van-Gieson staining and Masson trichrome staining of hepatic specimens in mice with Schistosoma japonicum infection. A model of hepatic granuloma and fibrosis was established by infecting mice with S. japonicum cercariae, then the hepatic specimens were taken and Van-Gieson staining and Masson trichrome staining were performed. Eventually, the area of granuloma and fibrosis were measured by imaging analysis software. When the time of staining was 3-7 min, there was no significant difference of the fibrosis areas between the two methods (P > 0.05); when the time of staining was more than 10 min, the staining area showed by Masson's staining was significantly larger than that showed by Van-Gieson staining, and the difference was statistically significant (P Masson trichrome staining, therefore Van-Gieson staining is a better method to display collagen.

  8. Bradyrhizobium valentinum sp. nov., isolated from effective nodules of Lupinus mariae-josephae, a lupine endemic of basic-lime soils in Eastern Spain.

    Science.gov (United States)

    Durán, David; Rey, Luis; Navarro, Albert; Busquets, Antonio; Imperial, Juan; Ruiz-Argüeso, Tomás

    2014-07-01

    Bacterial strains isolated from nitrogen-fixing nodules of Lupinus mariae-josephae have been characterized following genetic, phenotypic and symbiotic approaches. Analysis of 16S rRNA genes placed them in a group together with Bradyrhizobium elkanii USDA 76(T), B. pachyrhizi PAC48(T), B. jicamae PAC68(T), 'B. retamae' Ro19(T) and B. lablabi CCBAU 23086(T) with over 99.0% identity. Phylogenetic analysis of concatenated housekeeping genes, recA, atpD and glnII, suggested that L. mariae-josephae strains represent a new Bradyrhizobium species, closely related to B. lablabi CCBAU 23086(T), B. jicamae PAC68(T) and 'B. retamae' Ro19(T) with 92.1, 91.9 and 90.8% identity, respectively. These results are consistent with overall genomic identities calculated as Average Nucleotide Identity (ANIm) using draft genomic sequences obtained for relevant strains. While L. mariae-josephae strains LmjM3(T)/LmjM6 exhibited a 99.2% ANIm value, they were significantly distant (<93% ANIm) from type strains of their closest species ('B. retamae' Ro19(T), B. lablabi CCBAU 23086(T) and B. jicamae PAC68(T)). Whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (WC-MALDI-TOF-MS) analysis of proteomic patterns of the same strains was consistent with these results. The symbiosis-related genes nodC, nodA and nifH genes from strains nodulating L. mariae-josephae were phylogenetically related to those from 'B. retamae' Ro19(T), but divergent from those of strains that nodulate other lupine species. Based on genetic, genomic, proteomic and phenotypic data presented in this study, L. mariae-josephae nodulating strains LmjM3(T), LmjM6 and LmjM2 should be grouped within a new species for which the name Bradyrhizobium valentinum sp. nov. is proposed (type strain LmjM3(T)=CECT 8364(T), LMG 2761(T)). Copyright © 2014 Elsevier GmbH. All rights reserved.

  9. Clinical Effects of Formulated Food of Peucedanum japonicum Extract and Saw Palmetto Extract in Male Patients with Lower Urinary Tract Symptoms.

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    Kageyama, Shinji; Beppu, Masanori; Ohnogi, Hiromu; Miyazaki, Sayaka; Haruno, Akihiro; Ito, Yoshihiko; Yamada, Shizuo

    2017-02-04

    To evaluate changes over time in subjective symptom scores and urination parameters before and after oral administration of formulated food containing a combination of Peucedanum japonicum (P. japonicum) extract and saw palmetto extract (SPE) in male patients with lower urinary tract symptoms (LUTS). This study was conducted in an open label manner on male patients with untreated LUTS. The urination state of patients was evaluated before and after administration of food formulated with P. japonicum extract and SPE for 4 weeks, based on urodynamic parameters and subjective symptom scores (International Prostate Symptom Score [IPSS and IPSS-QOL], Overactive Bladder Symptom Score [OABSS], Overactive Bladder Questionnaire [OAB-q], and International Index of Erectile Function [IIEF]). After the administration of food formulated with these extracts, the following results were obtained: (i) Subjective findings: The IPSS-QOL score improved significantly; both parameters related to nocturia, i.e., frequency of nighttime urination and OABSS-2, improved significantly; other ratings for subjective symptoms slightly improved. (ii) Objective findings: Residual urine volume decreased significantly, and blood prostate specific antigen (PSA) and urinary 8-OHdG levels decreased slightly after the treatment. (iii) Other findings: Blood pressure decreased slightly. No adverse drug reactions were reported. (iv) Patient impressions: 75% of patients gave a rating of "Good" or higher, with 15 out of 20 patients wanting to continue treatment after the end of 4-week administration period. Food formulated with P. japonicum extract and SPE may be useful to decrease frequency of nighttime urination and residual urine volume in male patients with LUTS. © 2017 John Wiley & Sons Australia, Ltd.

  10. Genetic Structure Inferred from Mitochondrial 12S Ribosomal RNA Sequence of Oncomelania quadrasi, the Intermediate Snail Host of Schistosoma japonicum in the Philippines

    OpenAIRE

    Saijuntha, Weerachai; Jarilla, Blanca; Leonardo, Alvin K.; Sunico, Louie S.; Leonardo, Lydia R.; Andrews, Ross H.; Sithithaworn, Paiboon; Trevor N Petney; Kirinoki, Masashi; Kato-Hayashi, Naoko; Kikuchi, Mihoko; Chigusa, Yuichi; Agatsuma, Takeshi

    2014-01-01

    Species and subspecies of the Oncomelania hupensis species complex are recognized as intermediate hosts of Schistosoma japonicum. Of these species and subspecies, O. quadrasi is distributed throughout the Philippines. This study used 12S ribosomal RNA sequences to explore the genetic structure of O. quadrasi populations in the Philippines. Three subspecies, O. h. hupensis, O. h. formosana, and O. h. chiui of this group were also examined. The phylogenetic tree and haplotypes network showed th...

  11. CD4+CD25+ Treg induction by an HSP60-derived peptide SJMHE1 from Schistosoma japonicum is TLR2 dependent.

    Science.gov (United States)

    Wang, Xuefeng; Zhou, Sha; Chi, Ying; Wen, Xiaoyun; Hoellwarth, Jason; He, Lei; Liu, Feng; Wu, Calvin; Dhesi, Shawn; Zhao, Jiaqing; Hu, Wei; Su, Chuan

    2009-11-01

    Chronic schistosome infection results in the suppression of host immune responses, allowing long-term schistosome survival and restricting pathology. Current theories suggest that Treg play an important role in this regulation. However, the mechanism of Treg induction during schistosome infection is still unknown. The aim of this study was to determine the mechanism behind the induction of CD4(+)CD25(+) T cells by Schistosoma japonicum HSP60 (SjHSP60)-derived peptide SJMHE1 as well as to elucidate the cellular and molecular basis for the induction of CD4(+)CD25(+) T cells during S. japonicum infection. Mice immunized with SJMHE1 or spleen and LN cells from naïve mice pretreated with SJMHE1 in vitro all displayed an increase in CD4(+)CD25(+) T-cell populations. Release of IL-10 and TGF-beta by SJMHE1 stimulation may contribute to suppression. Adoptively transferred SJMHE1-induced CD4(+)CD25(+) T cells inhibited delayed-type hypersensitivity in BALB/c mice. Additionally, SJMHE1-treated APC were tolerogenic and induced CD4(+) cells to differentiate into suppressive CD4(+)CD25(+) Treg. Furthermore, our data support a role for TLR2 in SJMHE1-mediated CD4(+)CD25(+) Treg induction. These findings provide the basis for a more complete understanding of the S. japonicum-host interactions that contribute to host homeostatic mechanisms, preventing an excessive immune response.

  12. The dynamic changes of CD3e(-)CD11c(+) dendritic cells in spleens and bone marrow of mice infected with Schistosoma japonicum.

    Science.gov (United States)

    Chen, Lin; Chen, Qingzhou; Hou, Wei; He, Li

    2017-03-01

    Schistosoma japonicum as a pathogeny requires dendritic cells to activate immune response. So, the research is to study the dynamic changes of CD3e(-)CD11c(+) dendritic cells in mice infected with S. japonicum. Zero, 7, 28, 35, and 63 days were selected to study the variation of dendritic cells, and the proportions of CD3e(-)CD11c(+) dendritic cells and CD86(+) mature dendritic cells in spleens and bone marrow were tested by flow cytometry. As a result, the variation trends of dendritic cells in spleen and bone marrow are similar as follows: the proportions of CD3e(-)CD11c(+) dendritic cells increased first and then decreased from day 35, but the percentages of CD86(+) mature dendritic cells decreased from day 28 and increased in day 63. In vitro, cultured dendritic cells treated with SEA and SAWA were tested by flow cytometry, the variation trends of CD86 on dendritic cells are consistent with the results in days 28 and 63. Besides CD86, the expression of MHC-II also hints immune regulation. In conclusion, it is speculated that dendritic cells play a role of immune regulation through MHC-II and CD86 in S. japonicum infection. Immune regulation of dendritic cells is not only in favor of the survival of host and parasite but also can be used in the therapy for immune diseases.

  13. Eco-social determinants of Schistosoma japonicum infection supported by multi-level modelling in Eryuan county, People's Republic of China.

    Science.gov (United States)

    Yang, Kun; Zhou, Xiao-Nong; Jia, Tie-Wu; Yang, Guo-Jing; Wu, Xiao-Hua; Shi, Xue-Wen; Li, Hong-Jun; Steinmann, Peter; Utzinger, Jürg; Bergquist, Robert

    2015-01-01

    Schistosomiasis remains of considerable public health concern in many tropical and subtropical regions of the world, including the People's Republic of China (P.R. China). The effectiveness of schistosomiasis control interventions are, among other factors, governed by the social-ecological context. However, eco-social determinants of schistosomiasis are poorly understood, particularly at the household or village levels. In the current study, residents in 26 villages of Eryuan county, Yunnan province, P.R. China, were screened for Schistosoma japonicum infection with a serological assay that was followed by stool examination for sero-positive individuals. Bayesian multilevel models with spatial random effects were employed to profile the S. japonicum infection risk based on known transmission sites of S. japonicum that are scattered across individual land parcels in this part of the country. The key risk factors identified with this approach were the absence of a sanitary stall house for livestock and presence of living and infected intermediate host snails in close proximity. We conclude that a spatially explicit Bayesian multilevel approach can deepen our understanding of eco-social determinants that govern schistosomiasis transmission at a small geographical scale. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. High-quality permanent draft genome sequence of Bradyrhizobium sp. Ai1a-2; a microsymbiont of Andira inermis discovered in Costa Rica.

    Science.gov (United States)

    Tian, Rui; Parker, Matthew; Seshadri, Rekha; Reddy, Tbk; Markowitz, Victor; Ivanova, Natalia; Pati, Amrita; Woyke, Tanja; Baeshen, Mohammed; Baeshen, Nabih; Kyrpides, Nikos; Reeve, Wayne

    2015-01-01

    Bradyrhizobium sp. Ai1a-2 is is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from an effective nitrogen fixing root nodule of Andira inermis collected from Tres Piedras in Costa Rica. In this report we describe, for the first time, the genome sequence information and annotation of this legume microsymbiont. The 9,029,266 bp genome has a GC content of 62.56% with 247 contigs arranged into 246 scaffolds. The assembled genome contains 8,482 protein-coding genes and 102 RNA-only encoding genes. This rhizobial genome was sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project proposal.

  15. Bradyrhizobium-Lupinus mariae-josephae: a unique symbiosis endemic of a basic soil in Eastern Spain

    Science.gov (United States)

    Durán, D.; Sánchez-Cañizares, C.; Navarro, A.; Rey, L.; Imperial, J.; Ruiz-Argüeso, T.

    2012-04-01

    Lupinus mariae-josephae is an intriguing lupine species recently discovered in the Mediterranean region and constitutes an endemism of a small area of Eastern Spain (Valencia province; Pascual, 2004; Mahé et al. 2011). It opens new perspectives for ecological and agronomic interests, as it represents the sole lupine species that preferentially grows in basic soils, while almost all other lupine species occur in acid to neutral soils. The L. mariae-josephae symbionts isolated from soils of calcareous areas of Valencia are extremely slow-growing bacteria belonging to the Bradyrhrizobium genus and showing symbiotic specificity that prevents nodulation of other Lupinus spp. such as L. angustifolius or L. luteus typically thriving in acid soils (Sanchez-Cañizares et al, 2011). Their phylogenetic analysis based on housekeeping and symbiotic genes showed that L. mariae-josephae symbionts belong to an evolutionary lineage that also includes endosymbiotic bacteria from Retama spp. of Northern Algeria basic soils (Boulila et al. 2009). Conversely, this new lineage is phylogenetically distinct from that of endosymbiotic bacteria from other Lupinus spp. native of the Iberian Peninsula, which were nested mainly within B. canariense and B. japonicum lineages. A genomic diversity study of the indigenous bradyrhizobia population of the calcareous areas in Valencia, based on fingerprint and phylogenetic analysis, showed the existence of a large diversity of genotypes, some of which are related to bacteria from the Retama spp. symbiosis in Algeria. This singular genomic divergence of L. mariae-josephae symbiotic bacteria in such a small geographical area fosters attractive studies on the origin, ecology and evolution of both partners of the symbiosis. Furthermore, it is expected that ongoing seed inoculation experiments with selected strains will allow us to extend the extant distribution spots of L. mariae-josephae plants in Valencia area, and also to determine whether the

  16. The Type III Secretion System (T3SS) is a Determinant for Rice-Endophyte Colonization by Non-Photosynthetic Bradyrhizobium.

    Science.gov (United States)

    Piromyou, Pongdet; Songwattana, Pongpan; Greetatorn, Teerana; Okubo, Takashi; Kakizaki, Kaori Chiba; Prakamhang, Janpen; Tittabutr, Panlada; Boonkerd, Nantakorn; Teaumroong, Neung; Minamisawa, Kiwamu

    2015-01-01

    Plant associations by bradyrhizobia have been detected not only in leguminous plants, but also in non-leguminous species including rice. Bradyrhizobium sp. SUTN9-2 was isolated from Aeschynomene americana L., which is a leguminous weed found in the rice fields of Thailand. This strain promoted the highest total rice (Oryza sativa L. cultivar Pathum Thani 1) dry weight among the endophytic bradyrhizobial strains tested, and was, thus, employed for the further characterization of rice-Bradyrhizobium interactions. Some known bacterial genes involved in bacteria-plant interactions were selected. The expression of the type III secretion component (rhcJ), type IV secretion component (virD4), and pectinesterase (peces) genes of the bacterium were up-regulated when the rice root exudate was added to the culture. When SUTN9-2 was inoculated into rice seedlings, the peces, rhcJ, virD4, and exopolysaccharide production (fliP) genes were strongly expressed in the bacterium 6-24 h after the inoculation. The gene for glutathione-S-transferase (gst) was slightly expressed 12 h after the inoculation. In order to determine whether type III secretion system (T3SS) is involved in bradyrhizobial infections in rice plants, wild-type SUTN9-2 and T3SS mutant strains were inoculated into the original host plant (A. americana) and a rice plant (cultivar Pathum Thani 1). The ability of T3SS mutants to invade rice tissues was weaker than that of the wild-type strain; however, their phenotypes in A. americana were not changed by T3SS mutations. These results suggest that T3SS is one of the important determinants modulating rice infection; however, type IV secretion system and peces may also be responsible for the early steps of rice infection.

  17. Type 3 Secretion System (T3SS of Bradyrhizobium sp. DOA9 and Its Roles in Legume Symbiosis and Rice Endophytic Association

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    Pongpan Songwattana

    2017-09-01

    Full Text Available The Bradyrhizobium sp. DOA9 strain isolated from a paddy field has the ability to nodulate a wide spectrum of legumes. Unlike other bradyrhizobia, this strain has a symbiotic plasmid harboring nod, nif, and type 3 secretion system (T3SS genes. This T3SS cluster contains all the genes necessary for the formation of the secretory apparatus and the transcriptional activator (TtsI, which is preceded by a nod-box motif. An in silico search predicted 14 effectors putatively translocated by this T3SS machinery. In this study, we explored the role of the T3SS in the symbiotic performance of DOA9 by evaluating the ability of a T3SS mutant (ΩrhcN to nodulate legumes belonging to Dalbergioid, Millettioid, and Genistoid tribes. Among the nine species tested, four (Arachis hypogea, Vigna radiata, Crotalaria juncea, and Macroptilium atropurpureum responded positively to the rhcN mutation (ranging from suppression of plant defense reactions, an increase in the number of nodules and a dramatic improvement in nodule development and infection, one (Stylosanthes hamata responded negatively (fewer nodules and less nitrogen fixation and four species (Aeschynomene americana, Aeschynomene afraspera, Indigofera tinctoria, and Desmodium tortuosum displayed no phenotype. We also tested the role of the T3SS in the ability of the DOA9 strain to endophytically colonize rice roots, but detected no effect of the T3SS mutation, in contrast to what was previously reported in the Bradyrhizobium SUTN9-2 strain. Taken together, these data indicate that DOA9 contains a functional T3SS that interferes with the ability of the strain to interact symbiotically with legumes but not with rice.

  18. Type 3 Secretion System (T3SS) of Bradyrhizobium sp. DOA9 and Its Roles in Legume Symbiosis and Rice Endophytic Association.

    Science.gov (United States)

    Songwattana, Pongpan; Noisangiam, Rujirek; Teamtisong, Kamonluck; Prakamhang, Janpen; Teulet, Albin; Tittabutr, Panlada; Piromyou, Pongdet; Boonkerd, Nantakorn; Giraud, Eric; Teaumroong, Neung

    2017-01-01

    The Bradyrhizobium sp. DOA9 strain isolated from a paddy field has the ability to nodulate a wide spectrum of legumes. Unlike other bradyrhizobia, this strain has a symbiotic plasmid harboring nod, nif, and type 3 secretion system (T3SS) genes. This T3SS cluster contains all the genes necessary for the formation of the secretory apparatus and the transcriptional activator (TtsI), which is preceded by a nod-box motif. An in silico search predicted 14 effectors putatively translocated by this T3SS machinery. In this study, we explored the role of the T3SS in the symbiotic performance of DOA9 by evaluating the ability of a T3SS mutant (ΩrhcN) to nodulate legumes belonging to Dalbergioid, Millettioid, and Genistoid tribes. Among the nine species tested, four (Arachis hypogea, Vigna radiata, Crotalaria juncea, and Macroptilium atropurpureum) responded positively to the rhcN mutation (ranging from suppression of plant defense reactions, an increase in the number of nodules and a dramatic improvement in nodule development and infection), one (Stylosanthes hamata) responded negatively (fewer nodules and less nitrogen fixation) and four species (Aeschynomene americana, Aeschynomene afraspera, Indigofera tinctoria, and Desmodium tortuosum) displayed no phenotype. We also tested the role of the T3SS in the ability of the DOA9 strain to endophytically colonize rice roots, but detected no effect of the T3SS mutation, in contrast to what was previously reported in the Bradyrhizobium SUTN9-2 strain. Taken together, these data indicate that DOA9 contains a functional T3SS that interferes with the ability of the strain to interact symbiotically with legumes but not with rice.

  19. [Preliminary observation on the effect of mefloquine against egg granuloma formation in the liver of mice infected with Schistosoma japonicum].

    Science.gov (United States)

    Xiao, Shu-Hua; Zhang, Chao-Wei

    2013-04-01

    To explore whether mefloquine possesses the effect on granuloma formation induced by Schistosoma japonicum eggs. Seventeen out of twenty-eight mice infected with 20 S. japonicum cercariae for 35 days were treated orally with mefloquine at a single dose of 200 mg/kg, and groups of 2-3 mice were sacrificed at various intervals post-treatment. The livers removed from each group of mice were fixed in 10% formaldehyde. While the remained 11 untreated mice divided into 6 groups (1-2 mice per group) were sacrificed at the same time periods as groups of mice treated with mefloquine, and their livers served as untreated corresponding controls. The granulomas with single egg in the center were counted and their diameters were measured using an ocular micrometer. The liver tissue sections were stained with hematoxylin and eosin (HE), Foot's or Mallory's methods for observation on histopathological alteration of egg granulomas, and on the appearance of reticular and collagen fibers within the granulomas. After infected mice were treated with mefloquine for 3, 7, 14, 21, 28 and 35 days, i.e., 38, 42, 49, 56, 63, and 70 days post-infection, the mean diameters of granuloma with single egg measured in the liver tissues section were (161 +/- 19), (175 +/- 13), (195 +/- 9), (171 +/- 40), (180 +/- 13), and (145 +/- 25) microm, respectively, and each of them was significantly lower than that of its corresponding control group of (189 +/- 18), (197 +/- 11), (211 +/- 12), (208 +/- 19), (203 +/- 16), and (207 +/- 36) microm (P granuloma were sustained to 14-21 d post treatment (49-56 d post infection), which was significantly different from the corresponding control groups that all the eggs were surrounded by fibroblasts at 42 d post infection. Up to 28-35 d post treatment (63-70 d post infection), the boundary of egg granulomas distributed in the liver tissues of mefloquine treated groups was nearer in comparison to the corresponding control groups. Further observation on the reticular

  20. Green synthesis of silver nanoparticles using Ganoderma neo-japonicum Imazeki: a potential cytotoxic agent against breast cancer cells

    Science.gov (United States)

    Gurunathan, Sangiliyandi; Raman, Jegadeesh; Malek, Sri Nurestri Abd; John, Priscilla A; Vikineswary, Sabaratnam

    2013-01-01

    Background Silver nanoparticles (AgNPs) are an important class of nanomaterial for a wide range of industrial and biomedical applications. AgNPs have been used as antimicrobial and disinfectant agents due their detrimental effect on target cells. The aim of our study was to determine the cytotoxic effects of biologically synthesized AgNPs using hot aqueous extracts of the mycelia of Ganoderma neo-japonicum Imazeki on MDA-MB-231 human breast cancer cells. Methods We developed a green method for the synthesis of water-soluble AgNPs by treating silver ions with hot aqueous extract of the mycelia of G. neo-japonicum. The formation of AgNPs was characterized by ultraviolet-visible absorption spectroscopy, X-ray diffraction, dynamic light scattering, and transmission electron microscopy. Furthermore, the toxicity of synthesized AgNPs was evaluated using a series of assays: such as cell viability, lactate dehydrogenase leakage, reactive oxygen species generation, caspase 3, DNA laddering, and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling in human breast cancer cells (MDA-MB-231). Results The ultraviolet-visible absorption spectroscopy results showed a strong resonance centered on the surface of AgNPs at 420 nm. The X-ray diffraction analysis confirmed that the synthesized AgNPs were single-crystalline, corresponding with the result of transmission electron microscopy. Treatment of MDA-MB-231 breast cancer cells with various concentrations of AgNPs (1–10 μg/mL) for 24 hours revealed that AgNPs could inhibit cell viability and induce membrane leakage in a dose-dependent manner. Cells exposed to AgNPs showed increased reactive oxygen species and hydroxyl radical production. Furthermore, the apoptotic effects of AgNPs were confirmed by activation of caspase 3 and DNA nuclear fragmentation. Conclusion The results indicate that AgNPs possess cytotoxic effects with apoptotic features and suggest that the reactive oxygen species generated by

  1. Green synthesis of silver nanoparticles using Ganoderma neo-japonicum Imazeki: a potential cytotoxic agent against breast cancer cells.

    Science.gov (United States)

    Gurunathan, Sangiliyandi; Raman, Jegadeesh; Abd Malek, Sri Nurestri; John, Priscilla A; Vikineswary, Sabaratnam

    2013-01-01

    Silver nanoparticles (AgNPs) are an important class of nanomaterial for a wide range of industrial and biomedical applications. AgNPs have been used as antimicrobial and disinfectant agents due their detrimental effect on target cells. The aim of our study was to determine the cytotoxic effects of biologically synthesized AgNPs using hot aqueous extracts of the mycelia of Ganoderma neo-japonicum Imazeki on MDA-MB-231 human breast cancer cells. We developed a green method for the synthesis of water-soluble AgNPs by treating silver ions with hot aqueous extract of the mycelia of G. neo-japonicum. The formation of AgNPs was characterized by ultraviolet-visible absorption spectroscopy, X-ray diffraction, dynamic light scattering, and transmission electron microscopy. Furthermore, the toxicity of synthesized AgNPs was evaluated using a series of assays: such as cell viability, lactate dehydrogenase leakage, reactive oxygen species generation, caspase 3, DNA laddering, and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling in human breast cancer cells (MDA-MB-231). The ultraviolet-visible absorption spectroscopy results showed a strong resonance centered on the surface of AgNPs at 420 nm. The X-ray diffraction analysis confirmed that the synthesized AgNPs were single-crystalline, corresponding with the result of transmission electron microscopy. Treatment of MDA-MB-231 breast cancer cells with various concentrations of AgNPs (1-10 μg/mL) for 24 hours revealed that AgNPs could inhibit cell viability and induce membrane leakage in a dose-dependent manner. Cells exposed to AgNPs showed increased reactive oxygen species and hydroxyl radical production. Furthermore, the apoptotic effects of AgNPs were confirmed by activation of caspase 3 and DNA nuclear fragmentation. The results indicate that AgNPs possess cytotoxic effects with apoptotic features and suggest that the reactive oxygen species generated by AgNPs have a significant role in

  2. Efeito de fontes e doses de molibdênio na sobrevivência do Bradyrhizobium e na fixação biológica de nitrogênio em soja

    Directory of Open Access Journals (Sweden)

    Albino Ulisses Brigatto

    2001-01-01

    Full Text Available O molibdênio (Mo é essencial para a soja, por participar da enzima nitrogenase, sintetizada pelas bactérias durante o processo de fixação biológica do nitrogênio (FBN por simbiose. A aplicação do Mo nas sementes no momento da semeadura antecede a aplicação do inoculante. O contato do Mo com o inoculante prejudica o Bradyrhizobium e, conseqüentemente, a FBN. O presente trabalho teve como objetivo estudar alternativas de fornecer Mo para a soja e a FBN sem afetar a sobrevivência da bactéria, a nodulação e a eficiência do processo de FBN. Avaliou-se, in vitro, a capacidade das estirpes de Bradyrhizobium de crescerem em meio líquido contendo concentrações crescentes de diversas fontes de Mo, e, em casa de vegetação, os efeitos de doses crescentes de molibdato de sódio e de outras fontes de Mo na sobrevivência do Bradyrhizobium, na nodulação e na eficiência da FBN. As estirpes apresentaram tolerância distinta com relação a fontes e doses de Mo; a estirpe SEMIA 5080 apresentou o melhor crescimento entre elas. A aplicação de Mo nas sementes com o inoculante deve ser evitada, porque ele reduz o número de células de Bradyrhizobium, a nodulação e a FBN. As fontes de Mo mais eficientes para a FBN são molibdato de amônio, trióxido de Mo e as fontes comerciais Grap e Comol.

  3. Pteryxin: a coumarin in Peucedanum japonicum Thunb leaves exerts antiobesity activity through modulation of adipogenic gene network.

    Science.gov (United States)

    Nugara, Ruwani N; Inafuku, Masashi; Takara, Kensaku; Iwasaki, Hironori; Oku, Hirosuke

    2014-10-01

    Partially purified hexane phase (HP) of Peucedanum japonicum Thunb (PJT) was identified as an antiobesity candidate. However, the compound responsible for the antiobesity activity remained unknown. Thus, in this study we isolated the active compound, to determine the mechanisms related to antiobesity activity in vitro. The HP was fractionated, and the effect on the triacylglycerol (TG) content was evaluated in 3T3-L1 preadipocytes and HepG2 hepatocytes. On the basis of comprehensive spectroscopic analyses, the structure of the active compound was identified as pteryxin, a known compound in PJT. However, to our knowledge, its biological activities against obesity have not been reported previously. The dose-dependent effect on the TG content, and the gene expressions related to adipogenesis, fatty acid catabolism, energy expenditure, lipolysis, and lipogenesis due to pteryxin (10, 15, and 20 μg/mL) were examined in vitro. Pteryxin dose dependently suppressed TG content in both 3T3-L1 adipocytes (by 52.7%, 53.8%, and 57.4%, respectively; P 0.05) were up-regulated due to pteryxin. Our study demonstrated that pteryxin in PJT plays the key role in regulating the lipid metabolism-related gene network and improving energy production in vitro. Thus, the results suggest pteryxin as a new natural compound to be used as an antiobesity drug in the pharmaceutical industry. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. SjE16.7 activates macrophages and promotes Schistosoma japonicum egg-induced granuloma development.

    Science.gov (United States)

    Fang, Yan; Wu, Chenyun; Chen, Qing; Wu, Jianhua; Yang, Yang; Guo, Xiaokui; Chen, Guangjie; Wang, Zhaojun

    2015-09-01

    SjE16.7 is an egg-specific protein from Schistosoma japonicum that recruits neutrophils and initiates an inflammatory granuloma response in host tissue. However, since macrophages are known to be important regulators of egg granuloma formation we investigated the effect of SjE16.7 on this cell type. Here we report that SjE16.7 is a potent macrophage activator, inducing macrophage chemotaxis and stimulating cytokine production. Treatment of murine primary macrophages with SjE16.7 resulted in upregulation of both pro- and anti-inflammatory cytokines (IL-10, IL-12, IL-6 and TNF-α), as well as phosphorylation of mitogen-activated protein kinases (MAPKs). Moreover, SjE16.7 treatment increased MHC Class II expression on the surface of macrophages. Importantly, in vivo blockade of SjE16.7 significantly reduced egg-induced pathology, as a result of decreased leucocyte infiltration and reduced granuloma size. Our results suggest that SjE16.7 is an important pathogenic factor and a potential treatment target for this disease. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Identification of (L)-3-hydroxykynurenine O-sulfate in the buccal gland secretion of the parasitic lamprey, Lethenteron japonicum.

    Science.gov (United States)

    Odani, Shoji; Ito, Naoko; Hasegawa, Mai; Uchiumi, Toshio; Hase, Sumihiro

    2012-12-01

    Parasitic lampreys are known to secrete proteins having anticoagulant and vasodilator activities from the buccal glands during feeding on their host's blood. However, small molecules in the secretion have never been explored in detail. We examined the secretion of Japanese liver lamprey (Lethenteron japonicum) for small molecules and found an intensely fluorescent substance upon gel filtration. After purification by anion-exchange chromatography and reversed-phase HPLC, structure of the compound was determined to be L-3-hydroxykynurenine O-sulfate by NMR- and UV-spectrometry, complemented with enzymatic and chemical degradation. In vertebrates, the sulfate ester of 3-hydroxykynurenine is a compound that has been regarded as a urinary metabolite of tryptophan but not reported from normal tissues to date. Although the function of this molecule in the buccal glands remains to be elucidated, it is remarkable that the same substance was described in 1960s from two species of blood-sucking insects, Rhodnius prolixus and Triatoma infestans, suggesting its potential role in blood-feeding.

  6. Oral delivery of the Sj23LHD-GST antigen by Salmonella typhimurium type III secretion system protects against Schistosoma japonicum infection in mice.

    Directory of Open Access Journals (Sweden)

    Guo Chen

    2011-09-01

    Full Text Available BACKGROUND: Schistosomiasis japonica is a zoonotic parasitic disease and oral vaccine delivery system would be benefit for prevention of this disease. Although attenuated salmonella has been used as an antigen expression vector for oral vaccine development, the membrane-bound vacuoles in which bacteria reside hinders the presentation of expressed heterologous antigens to the major histocompatibility complex (MHC molecules. The present work used an attenuated Salmonella typhimurium strain VNP20009 to secretory expression of Sj23LHDGST bivalent antigen from Schistosoma japonicum and tested the protective efficacy against S. japonicum infection in orally immunized mice. METHODOLOGY/PRINCIPAL FINDINGS: Promoters (nirB or pagC were used to express the antigen (Sj23LHDGST and the Salmonella type III or α-hemolysin secretion system was employed to secrete it. The immunoblotting analysis and fluorescent microscopy revealed that the antigen was effectively expressed and delivered to the cytosol of macrophages in vitro. Among recombinant vaccine strains, an engineered VNP20009 which expressed the antigen by nirB promoter and secreted it through type III secretion system (nirB-sopE(1-104-Sj23LHD-GST efficiently protected against S. japonicum infection in a mouse model. This strain elicited a predominantly IgG(2a antibody response and a markedly increase in the production of IL-12 and IFN-γ. The flow cytometric analysis demonstrated that this strain caused T cell activation as evidenced by significantly increased expression of CD44 and CD69. CONCLUSION/SIGNIFICANCE: Oral delivery of antigen by nirB-driven Salmonella typhimurium type III secretion system is a novel, safe, inexpensive, efficient and convenient approach for schistosome vaccine development.

  7. [Dynamic alteration of CD154/CD40 and its effects on Th1/Th2 polarization in inducible co-stimulator ligand knockout mice infected with Schistosoma japonicum].

    Science.gov (United States)

    Wang, Yu; Xia, Chao-ming

    2015-12-18

    To analyze effect on the CD154-CD40 signaling pathway and Th1/Th2 polarization by deficient inducible co-stimulator (ICOS)-ICOS ligand (ICOSL) signaling in mice infected with Schistosoma japonicum. ICOSL knockout (ICOSL-KO) mice and wild-type C57BL/6J mice were used as experimental Schistosomiasis model infected with Schistosoma japonicum. The expressions of CD154 and CD40 on splenocytes and on inflammatory cells around granulomatous infiltration of liver in mice infected with Schistosoma japonicum were analyzed by flow cytometry,immunohistochemical staining, respectively, on the day before infection (0 week)and at the end of 4, 7, 12, 16 and 20 weeks post-infection. The splenocytes of the mice were stimulated with soluble egg antigen(SEA) for 72 hours, then the concentrations of interferon gamma(IFN-γ) and interleukin-4 (IL-4) in the culture supernatants were measured by sandwich enzyme-linked immunosorbent assay (ELISA) kits. The levels of SEA-specific antibodies of IgG and IgG1 and IgG2a were measured in the mice sera by ELISA. The granulomatous pathology in the mice liver was dynamically observed by hematoxylin and eosin (HE) staining. Compared with the wild-type C57BL/6J mice, the expressions of CD154 on CD4+ T splenocytes [(18.62 ± 4.76)% vs.(27.91 ± 3.94)%, (22.44 ± 4.67)% vs.(40.86 ± 5.21)%, (25.50 ± 6.81)% vs.(43.81 ± 8.41)%, (20.22 ± 5.28)% vs.(40.95 ± 7.34)%, (17.87 ± 4.59)% vs.(33.16 ± 6.31)%, all PCD154[(0.319 ± 0.066) vs.(0.488 ± 0.086), (0.389 ± 0.067) vs.(0.596 ± 0.082), (0.378 ± 0.064) vs.(0.543 ± 0.072), (0.348 ± 0.069) vs.(0.523 ± 0.076), all PCD154 and CD40 and impairment of Th2 immune response in the ICOSL-KO mice infected with Schistosoma japonicum, accompanying with notedly reduced hepatic granulomatous pathology. The ICOS-ICOSL signaling has a regulatory effect on CD154-CD40 signaling pathway, and may play an important role in the hepatic egg granuloma formation of Schistosomiasis.

  8. An investigation into the potential effects of infrapopulation structure and other sources of sampling error, on population genetic studies of the transmission of Schistosoma japonicum (Trematoda: Digenea).

    Science.gov (United States)

    Huo, Guan-Nan; Liu, Liang; He, Hong-Bin; Attwood, Stephen W

    2016-03-21

    Schistosoma japonicum remains a major challenge to human and animal health. Earlier microsatellite-based studies reported possible definitive-host-specific private alleles within S. japonicum, opening the possibility that different definitive hosts might harbour different parasite strains. Previous investigations have also detected near-identical multilocus genotypes in populations of adult worms - possibly the result of mutations occurring during the asexual (intramolluscan) phase of clonal expansion. Research has also revealed extensive deviations from Hardy-Weinberg Proportions (HWP) and conflicting results among studies. The present study was performed to examine some of the potential effects of infrapopulation structure on microsatellite-based studies of the transmission ecology of S. japonicum. Potential sources of bias considered included organotropic distribution of worms, non-random mating and corrections for clonal expansion. Stool samples from naturally infected hosts were used to infect snails in the laboratory and thereby expose mice. 274 individual worms were typed at seven microsatellite loci. Removal of individuals bearing duplicate MLGs (as a correction for presumed clonal expansion) had an impact on both HWP and organotropic genetic differentiation. The study found no evidence that heterozygote deficiencies were caused by a Wahlund effect. Female-male pairings appeared to be random and there was no evidence for mate choice by heterozygosity. There was some indication that excess heterozygosity, induced by clonal expansion, can offset heterozygote deficiencies caused by small population size or populations fragmented by parasite control efforts. The view is supported that miracidia are preferable to adult worms in investigations into host-specific parasite lineages. Where adults must be used, extreme care should be taken with regard to sampling if infrapopulations of small animals are compared with those of larger animals; this is because of

  9. High-quality permanent draft genome sequence of the Bradyrhizobium elkanii type strain USDA 76(T), isolated from Glycine max (L.) Merr.

    Science.gov (United States)

    Reeve, Wayne; van Berkum, Peter; Ardley, Julie; Tian, Rui; Gollagher, Margaret; Marinova, Dora; Elia, Patrick; Reddy, T B K; Pillay, Manoj; Varghese, Neha; Seshadri, Rekha; Ivanova, Natalia; Woyke, Tanja; Baeshen, Mohamed N; Baeshen, Nabih A; Kyrpides, Nikos

    2017-01-01

    Bradyrhizobium elkanii USDA 76(T) (INSCD = ARAG00000000), the type strain for Bradyrhizobium elkanii, is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from an effective nitrogen-fixing root nodule of Glycine max (L. Merr) grown in the USA. Because of its significance as a microsymbiont of this economically important legume, B. elkanii USDA 76(T) was selected as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria sequencing project. Here the symbiotic abilities of B. elkanii USDA 76(T) are described, together with its genome sequence information and annotation. The 9,484,767 bp high-quality draft genome is arranged in 2 scaffolds of 25 contigs, containing 9060 protein-coding genes and 91 RNA-only encoding genes. The B. elkanii USDA 76(T) genome contains a low GC content region with symbiotic nod and fix genes, indicating the presence of a symbiotic island integration. A comparison of five B. elkanii genomes that formed a clique revealed that 356 of the 9060 protein coding genes of USDA 76(T) were unique, including 22 genes of an intact resident prophage. A conserved set of 7556 genes were also identified for this species, including genes encoding a general secretion pathway as well as type II, III, IV and VI secretion system proteins. The type III secretion system has previously been characterized as a host determinant for Rj and/or rj soybean cultivars. Here we show that the USDA 76(T) genome contains genes encoding all the type III secretion system components, including a translocon complex protein NopX required for the introduction of effector proteins into host cells. While many bradyrhizobial strains are unable to nodulate the soybean cultivar Clark (rj1), USDA 76(T) was able to elicit nodules on Clark (rj1), although in reduced numbers, when plants were grown in Leonard jars containing sand or vermiculite. In these conditions, we postulate that the presence of Nop

  10. Comparative Analysis of Transcriptional Profiles of Adult Schistosoma japonicum from Different Laboratory Animals and the Natural Host, Water Buffalo

    Science.gov (United States)

    Wu, Chuang; Hou, Nan; Chen, Qijun

    2015-01-01

    Background Schistosomiasis is one of the most widely distributed parasitic diseases in the world. Schistosoma japonicum, a zoonotic parasite with a wide range of mammalian hosts, is one of the major pathogens of this disease. Although numerous studies on schistosomiasis japonica have been performed using laboratory animal models, systematic comparative analysis of whole-genome expression profiles in parasites from different laboratory animals and nature mammalian hosts is lacking to date. Methodology/Principal Findings Adult schistosomes were obtained from laboratory animals BALB/c mice, C57BL/6 mice, New Zealand white rabbits and the natural host, water buffaloes. The gene expression profiles of schistosomes from these animals were obtained and compared by genome-wide oligonucleotide microarray analysis. The results revealed that the gene expression profiles of schistosomes from different laboratory animals and buffaloes were highly consistent (r>0.98) genome-wide. Meanwhile, a total of 450 genes were identified to be differentially expressed in schistosomes which can be clustered into six groups. Pathway analysis revealed that these genes were mainly involved in multiple signal transduction pathways, amino acid, energy, nucleotide and lipid metabolism. We also identified a group of 1,540 abundantly and stably expressed gene products in adult worms, including a panel of 179 Schistosoma- or Platyhelminthes-specific genes that may be essential for parasitism and may be regarded as novel potential anti-parasite intervention targets for future research. Conclusions/Significance This study provides a comprehensive database of gene expression profiles of schistosomes derived from different laboratory animals and water buffaloes. An expanded number of genes potentially affecting the development of schistosomes in different animals were identified. These findings lay the foundation for schistosomiasis research in different laboratory animals and natural hosts at the

  11. Comparative Analysis of Transcriptional Profiles of Adult Schistosoma japonicum from Different Laboratory Animals and the Natural Host, Water Buffalo.

    Science.gov (United States)

    Liu, Shuai; Zhou, Xiaosu; Piao, Xianyu; Wu, Chuang; Hou, Nan; Chen, Qijun

    2015-08-01

    Schistosomiasis is one of the most widely distributed parasitic diseases in the world. Schistosoma japonicum, a zoonotic parasite with a wide range of mammalian hosts, is one of the major pathogens of this disease. Although numerous studies on schistosomiasis japonica have been performed using laboratory animal models, systematic comparative analysis of whole-genome expression profiles in parasites from different laboratory animals and nature mammalian hosts is lacking to date. Adult schistosomes were obtained from laboratory animals BALB/c mice, C57BL/6 mice, New Zealand white rabbits and the natural host, water buffaloes. The gene expression profiles of schistosomes from these animals were obtained and compared by genome-wide oligonucleotide microarray analysis. The results revealed that the gene expression profiles of schistosomes from different laboratory animals and buffaloes were highly consistent (r>0.98) genome-wide. Meanwhile, a total of 450 genes were identified to be differentially expressed in schistosomes which can be clustered into six groups. Pathway analysis revealed that these genes were mainly involved in multiple signal transduction pathways, amino acid, energy, nucleotide and lipid metabolism. We also identified a group of 1,540 abundantly and stably expressed gene products in adult worms, including a panel of 179 Schistosoma- or Platyhelminthes-specific genes that may be essential for parasitism and may be regarded as novel potential anti-parasite intervention targets for future research. This study provides a comprehensive database of gene expression profiles of schistosomes derived from different laboratory animals and water buffaloes. An expanded number of genes potentially affecting the development of schistosomes in different animals were identified. These findings lay the foundation for schistosomiasis research in different laboratory animals and natural hosts at the transcriptional level and provide a valuable resource for screening anti

  12. Comparative Analysis of Transcriptional Profiles of Adult Schistosoma japonicum from Different Laboratory Animals and the Natural Host, Water Buffalo.

    Directory of Open Access Journals (Sweden)

    Shuai Liu

    2015-08-01

    Full Text Available Schistosomiasis is one of the most widely distributed parasitic diseases in the world. Schistosoma japonicum, a zoonotic parasite with a wide range of mammalian hosts, is one of the major pathogens of this disease. Although numerous studies on schistosomiasis japonica have been performed using laboratory animal models, systematic comparative analysis of whole-genome expression profiles in parasites from different laboratory animals and nature mammalian hosts is lacking to date.Adult schistosomes were obtained from laboratory animals BALB/c mice, C57BL/6 mice, New Zealand white rabbits and the natural host, water buffaloes. The gene expression profiles of schistosomes from these animals were obtained and compared by genome-wide oligonucleotide microarray analysis. The results revealed that the gene expression profiles of schistosomes from different laboratory animals and buffaloes were highly consistent (r>0.98 genome-wide. Meanwhile, a total of 450 genes were identified to be differentially expressed in schistosomes which can be clustered into six groups. Pathway analysis revealed that these genes were mainly involved in multiple signal transduction pathways, amino acid, energy, nucleotide and lipid metabolism. We also identified a group of 1,540 abundantly and stably expressed gene products in adult worms, including a panel of 179 Schistosoma- or Platyhelminthes-specific genes that may be essential for parasitism and may be regarded as novel potential anti-parasite intervention targets for future research.This study provides a comprehensive database of gene expression profiles of schistosomes derived from different laboratory animals and water buffaloes. An expanded number of genes potentially affecting the development of schistosomes in different animals were identified. These findings lay the foundation for schistosomiasis research in different laboratory animals and natural hosts at the transcriptional level and provide a valuable resource

  13. Isojacareubin from the Chinese Herb Hypericum japonicum: Potent Antibacterial and Synergistic Effects on Clinical Methicillin-Resistant Staphylococcus aureus (MRSA)

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    Zuo, Guo-Ying; An, Jing; Han, Jun; Zhang, Yun-Ling; Wang, Gen-Chun; Hao, Xiao-Yan; Bian, Zhong-Qi

    2012-01-01

    Through bioassay-guided fractionation of the extracts from the aerial parts of the Chinese herb Hypericum japonicum Thunb. Murray, Isojacareubin (ISJ) was characterized as a potent antibacterial compound against the clinical methicillin-resistant Staphylococcus aureus (MRSA). The broth microdilution assay was used to determine the minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) of ISJ alone. The results showed that its MICs/MBCs ranged from 4/16 to 16/64 μg/mL, with the concentrations required to inhibit or kill 50% of the strains (MIC50/MBC50) at 8/16 μg/mL. Synergistic evaluations of this compound with four conventional antibacterial agents representing different types were performed by the chequerboard and time-kill tests. The chequerboard method showed significant synergy effects when ISJ was combined with Ceftazidime (CAZ), Levofloxacin (LEV) and Ampicillin (AMP), with the values of 50% of the fractional inhibitory concentration indices (FICI50) at 0.25, 0.37 and 0.37, respectively. Combined bactericidal activities were also observed in the time-kill dynamic assay. The results showed the ability of ISJ to reduce MRSA viable counts by log10CFU/mL at 24 h of incubation at a concentration of 1 × MIC were 1.5 (LEV, additivity), 0.92 (CAZ, indifference) and 0.82 (AMP, indifference), respectively. These in vitro anti-MRSA activities of ISJ alone and its synergy with conventional antibacterial agents demonstrated that ISJ enhanced their efficacy, which is of potential use for single and combinatory therapy of patients infected with MRSA. PMID:22942699

  14. Isojacareubin from the Chinese herb Hypericum japonicum: potent antibacterial and synergistic effects on clinical methicillin-resistant Staphylococcus aureus (MRSA).

    Science.gov (United States)

    Zuo, Guo-Ying; An, Jing; Han, Jun; Zhang, Yun-Ling; Wang, Gen-Chun; Hao, Xiao-Yan; Bian, Zhong-Qi

    2012-01-01

    Through bioassay-guided fractionation of the extracts from the aerial parts of the Chinese herb Hypericum japonicum Thunb. Murray, Isojacareubin (ISJ) was characterized as a potent antibacterial compound against the clinical methicillin-resistant Staphylococcus aureus (MRSA). The broth microdilution assay was used to determine the minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) of ISJ alone. The results showed that its MICs/MBCs ranged from 4/16 to 16/64 μg/mL, with the concentrations required to inhibit or kill 50% of the strains (MIC(50)/MBC(50)) at 8/16 μg/mL. Synergistic evaluations of this compound with four conventional antibacterial agents representing different types were performed by the chequerboard and time-kill tests. The chequerboard method showed significant synergy effects when ISJ was combined with Ceftazidime (CAZ), Levofloxacin (LEV) and Ampicillin (AMP), with the values of 50% of the fractional inhibitory concentration indices (FICI(50)) at 0.25, 0.37 and 0.37, respectively. Combined bactericidal activities were also observed in the time-kill dynamic assay. The results showed the ability of ISJ to reduce MRSA viable counts by log(10)CFU/mL at 24 h of incubation at a concentration of 1 × MIC were 1.5 (LEV, additivity), 0.92 (CAZ, indifference) and 0.82 (AMP, indifference), respectively. These in vitro anti-MRSA activities of ISJ alone and its synergy with conventional antibacterial agents demonstrated that ISJ enhanced their efficacy, which is of potential use for single and combinatory therapy of patients infected with MRSA.

  15. Schistosoma japonicum HSP60-derived peptide SJMHE1 suppresses delayed-type hypersensitivity in a murine model.

    Science.gov (United States)

    Wang, Xuefeng; Wang, Jun; Liang, Yong; Ni, Hongchang; Shi, Liang; Xu, Chengcheng; Zhou, Yuepeng; Su, Yuting; Mou, Xiao; Chen, Deyu; Mao, Chaoming

    2016-03-12

    Parasite-derived molecules with immunomodulatory properties, which have been optimised during host-parasite co-evolution, exhibit potential applications as novel immunotherapeutics. We have previously demonstrated that Schistosoma japonicum HSP60-derived peptide SJMHE1 induces CD4(+)CD25(+) regulatory T-cells (Tregs) and that adoptively transferred SJMHE1-induced CD4(+)CD25(+) Tregs inhibit delayed-type hypersensitivity (DTH) in mice. However, multiple concerns regarding this method render this treatment unsuitable. To gain further insights into the potential effects of SJMHE1, we used ovalbumin (OVA)-induced DTH and evaluated the effect of SJMHE1 on DTH mice. BALB/c mice were sensitised with OVA alone or combined with SJMHE1 and then challenged with OVA to induce DTH. We first analysed the potential effects of SJMHE1 by measuring DTH responses, T-cell responses, cytokine secretion, and Treg proportions. We then evaluated the expression levels of IL-10 and TGF-β1 in CD4(+)CD25(+) T-cells during DTH and Treg generation to identify the mechanism by which SJMHE1 suppresses DTH. SJMHE1 modulated the effector response against OVA-induced DTH and stimulated the production of the anti-inflammatory cytokines IL-10 and TGF-β1 in immunised mice through a mechanism involving CD4(+)CD25(+) Tregs. SJMHE1-induced CD4(+)CD25(+) Tregs expressed high levels of CTLA-4, IL-10, and TGF-β1, which substantially contributed to the suppressive activity during DTH. The administration of SJMHE1 to DTH in mice led to the expansion of CD4(+)CD25(+) Tregs from CD4(+)CD25(-) T-cells in the periphery, which inhibited DTH responses. Our study proves that the parasite-driven peptide suppresses DTH in mice, which may confer a new option for inflammation treatment.

  16. Symbiotic N2-Fixation Estimated by the 15N Tracer Technique and Growth of Pueraria phaseoloides (Roxb. Benth. Inoculated with Bradyrhizobium Strain in Field Conditions

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    Papa Saliou Sarr

    2016-01-01

    Full Text Available This field experiment was established in Eastern Cameroon to examine the effect of selected rhizobial inoculation on N2-fixation and growth of Pueraria phaseoloides. Treatments consisted of noninoculated and Bradyrhizobium yuanmingense S3-4-inoculated Pueraria with three replications each. Ipomoea batatas as a non-N2-fixing reference was interspersed in each Pueraria plot. All the twelve plots received 2 gN/m2 of 15N ammonium sulfate 10% atom excess. At harvest, dry matter yields and the nitrogen derived from atmospheric N2-fixation (%Ndfa of inoculated Pueraria were significantly (P<0.05 higher (81% and 10.83%, resp. than those of noninoculated Pueraria. The inoculation enhanced nodule dry weight 2.44-fold. Consequently, the harvested N significantly (P<0.05 increased by 83% in inoculated Pueraria, resulting from the increase in N2-fixation and soil N uptake. A loss of 55 to 60% of the N fertilizer was reported, and 36 to 40% of it was immobilized in soil. Here, we demonstrated that both N2-fixing potential of P. phaseoloides and soil N uptake are improved through field inoculations using efficient bradyrhizobial species. In practice, the inoculation contributes to maximize N input in soils by the cover crop’s biomass and represent a good strategy to improve soil fertility for subsequent cultivation.

  17. Inoculation of Mimosa latispinosa Lam with the Commercial Arbuscular Mycorrhizal Fungus Rhizophagus irregularis DAOM 197198, and Bradyrhizobium spp. Under Nursery Production Conditions in South-East Madagascar

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    Sarasin, G.

    2017-01-01

    Full Text Available Qit Madagascar Minerals (QMM has planned several actions to reduce the environmental footprint of its mining project located near the city of Fort-Dauphin (Madagascar. One of these actions is the reclamation of a portion of its mined sites. Different symbiotic strains were tested as bio-enhancers for the ecological restoration using Mimosa latispinosa Lam, a native and pioneer shrub. The symbiotic strains tested in nursery were the commercial strain of arbuscular mycorrhizal fungus, Rhizophagus irregularis DAOM197198, and two local strains of Bradyrhizobium spp., STM1415 and STM1447, inoculated alone or dually with the arbuscular mycorrhiza. Treatments did not significantly increase the plant height and dry mass. However, plants grown in tyndallized soil had better growth than those in unsterilized soil. Results obtained twenty weeks after inoculation suggest that soil tyndallization (heating at 100°C and at atmospheric pressure of 700 kPa during three hours is an effective method for nursery production of high quality seedlings of M. latispinosa.

  18. FixK2 Is the Main Transcriptional Activator of Bradyrhizobium diazoefficiens nosRZDYFLX Genes in Response to Low Oxygen

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    María J. Torres

    2017-08-01

    Full Text Available The powerful greenhouse gas, nitrous oxide (N2O has a strong potential to drive climate change. Soils are the major source of N2O and microbial nitrification and denitrification the main processes involved. The soybean endosymbiont Bradyrhizobium diazoefficiens is considered a model to study rhizobial denitrification, which depends on the napEDABC, nirK, norCBQD, and nosRZDYFLX genes. In this bacterium, the role of the regulatory cascade FixLJ-FixK2-NnrR in the expression of napEDABC, nirK, and norCBQD genes involved in N2O synthesis has been previously unraveled. However, much remains to be discovered regarding the regulation of the respiratory N2O reductase (N2OR, the key enzyme that mitigates N2O emissions. In this work, we have demonstrated that nosRZDYFLX genes constitute an operon which is transcribed from a major promoter located upstream of the nosR gene. Low oxygen was shown to be the main inducer of expression of nosRZDYFLX genes and N2OR activity, FixK2 being the regulatory protein involved in such control. Further, by using an in vitro transcription assay with purified FixK2 protein and B. diazoefficiens RNA polymerase we were able to show that the nosRZDYFLX genes are direct targets of FixK2.

  19. Effect of Trichoderma harzianum biomass and Bradyrhizobium sp. strain NC 92 to control leaf blight disease of bambara groundnut (Vigna subterranea caused by Rhizoctonia solani in the field

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    Mana Kanjanamaneesathian

    2007-01-01

    Full Text Available Four hundred and sixty two strains of Trichoderma spp. were isolated from 23 soil samples in which groundnut (Arachis hypogaea L. and bambara groundnut (Vigna subterranea L. had been planted in Songkhla, Phattalung, Nakhon Si Thammarat, Narathiwat and Yala provinces. These fungi were tested against Rhizoctonia solani, a causal agent of leaf blight of bambara groundnut, using dual culture technique on PDA medium. Among 462 isolates tested, 226 isolates had an ability to overgrow R. solani completely. Further testing found 13 isolates having the ability to parasitize mycelia of R. solani. Among these isolates, ThB-1-54 produced a cellulolytic enzyme on congo-red agar. This isolate was later identified as T. harzianum Rifai. In the field test, applying biomass of the isolate ThB-1-54 cultured on ground mesocarp fiber of oil palm, the combination of the isolate ThB-1-54 on ground mesocarp fiber of oil palm and Bradyrhizobium sp. (strain NC 92, or fungicide (iprodione had no effect on disease severity, yield, or the amount of total nitrogen content in stems or seeds of bambara groundnut plant.

  20. Simultaneous interaction of Arabidopsis thaliana with Bradyrhizobium Sp. strain ORS278 and Pseudomonas syringae pv. tomato DC3000 leads to complex transcriptome changes.

    Science.gov (United States)

    Cartieaux, Fabienne; Contesto, Céline; Gallou, Adrien; Desbrosses, Guilhem; Kopka, Joachim; Taconnat, Ludivine; Renou, Jean-Pierre; Touraine, Bruno

    2008-02-01

    Induced systemic resistance (ISR) is a process elicited by telluric microbes, referred to as plant growth-promoting rhizobacteria (PGPR), that protect the host plant against pathogen attacks. ISR has been defined from studies using Pseudomonas strains as the biocontrol agent. Here, we show for the first time that a photosynthetic Bradyrhizobium sp. strain, ORS278, also exhibits the ability to promote ISR in Arabidopsis thaliana, indicating that the ISR effect may be a widespread ability. To investigate the molecular bases of this response, we performed a transcriptome analysis designed to reveal the changes in gene expression induced by the PGPR, the pathogen alone, or by both. The results confirm the priming pattern of ISR described previously, meaning that a set of genes, of which the majority was predicted to be influenced by jasmonic acid or ethylene, was induced upon pathogen attack when plants were previously colonized by PGPR. The analysis and interpretation of transcriptome data revealed that 12-oxo-phytodienoic acid, an intermediate of the jasmonic acid biosynthesis pathway, is likely to be an actor in the signaling cascade involved in ISR. In addition, we show that the PGPR counterbalanced the pathogen-induced changes in expression of a series of genes.

  1. Integrated roles of BclA and DD-carboxypeptidase 1 in Bradyrhizobium differentiation within NCR-producing and NCR-lacking root nodules.

    Science.gov (United States)

    Barrière, Quentin; Guefrachi, Ibtissem; Gully, Djamel; Lamouche, Florian; Pierre, Olivier; Fardoux, Joël; Chaintreuil, Clémence; Alunni, Benoît; Timchenko, Tatiana; Giraud, Eric; Mergaert, Peter

    2017-08-22

    Legumes harbor in their symbiotic nodule organs nitrogen fixing rhizobium bacteria called bacteroids. Some legumes produce Nodule-specific Cysteine-Rich (NCR) peptides in the nodule cells to control the intracellular bacterial population. NCR peptides have antimicrobial activity and drive bacteroids toward terminal differentiation. Other legumes do not produce NCR peptides and their bacteroids are not differentiated. Bradyrhizobia, infecting NCR-producing Aeschynomene plants, require the peptide uptake transporter BclA to cope with the NCR peptides as well as a specific peptidoglycan-modifying DD-carboxypeptidase, DD-CPase1. We show that Bradyrhizobium diazoefficiens strain USDA110 forms undifferentiated bacteroids in NCR-lacking soybean nodules. Unexpectedly, in Aeschynomene afraspera nodules the nitrogen fixing USDA110 bacteroids are hardly differentiated despite the fact that this host produces NCR peptides, suggesting that USDA110 is insensitive to the host peptide effectors and that nitrogen fixation can be uncoupled from differentiation. In agreement with the absence of bacteroid differentiation, USDA110 does not require its bclA gene for nitrogen fixing symbiosis with these two host plants. Furthermore, we show that the BclA and DD-CPase1 act independently in the NCR-induced morphological differentiation of bacteroids. Our results suggest that BclA is required to protect the rhizobia against the NCR stress but not to induce the terminal differentiation pathway.

  2. Optimisation of a droplet digital PCR assay for the diagnosis of Schistosoma japonicum infection: A duplex approach with DNA binding dye chemistry.

    Science.gov (United States)

    Weerakoon, Kosala G; Gordon, Catherine A; Gobert, Geoffrey N; Cai, Pengfei; McManus, Donald P

    2016-06-01

    Schistosomiasis is a chronically debilitating helminth infection with a significant socio-economic and public health impact. Accurate diagnostics play a pivotal role in achieving current schistosomiasis control and elimination goals. However, many of the current diagnostic procedures, which rely on detection of schistosome eggs, have major limitations including lack of accuracy and the inability to detect pre-patent infections. DNA-based detection methods provide a viable alternative to the current tests commonly used for schistosomiasis diagnosis. Here we describe the optimisation of a novel droplet digital PCR (ddPCR) duplex assay for the diagnosis of Schistosoma japonicum infection which provides improved detection sensitivity and specificity. The assay involves the amplification of two specific and abundant target gene sequences in S. japonicum; a retrotransposon (SjR2) and a portion of a mitochondrial gene (nad1). The assay detected target sequences in different sources of schistosome DNA isolated from adult worms, schistosomules and eggs, and exhibits a high level of specificity, thereby representing an ideal tool for the detection of low levels of parasite DNA in different clinical samples including parasite cell free DNA in the host circulation and other bodily fluids. Moreover, being quantitative, the assay can be used to determine parasite infection intensity and, could provide an important tool for the detection of low intensity infections in low prevalence schistosomiasis-endemic areas. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Gene Gun Bombardment with DNA-Coated Golden Particles Enhanced the Protective Effect of a DNA Vaccine Based on Thioredoxin Glutathione Reductase of Schistosoma japonicum

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    Yan Cao

    2013-01-01

    Full Text Available Schistosomiasis, caused by infection with Schistosoma species, remains an important parasitic zoonosis. Thioredoxin glutathione reductase of Schistosoma japonicum (SjTGR plays an important role in the development of the parasite and for its survival. Here we present a recombinant plasmid DNA vaccine, pVAX1/SjTGR, to estimate its protection against S. japonicum in BALB/c mice. The DNA vaccine administrated by particle bombardment induced higher protection than by intramuscular injection. All animals vaccinated with pVAX1/SjTGR developed significant specific anti-SjTGR antibodies than control groups. Moreover, animals immunized by gene gun exhibited a splenocyte proliferative response, with an increase in IFN-γ and IL-4. The recombinant plasmid administrated by gene gun achieved a medium protective efficacy of 27.83–38.83% ( of worm reduction and 40.38–44.51% ( of liver egg count reduction. It suggests that different modes of administering a DNA vaccine can influence the protective efficacy induced by the vaccine. Interestingly, from the enzymatic activity results, we found that worms obtained from pVAX1/SjTGR-vaccinated animals expressed lower enzymatic activity than the control group and the antibodies weakened the enzymatic activity of SjTGR in vitro, too. It implies that the high-level antibodies may contribute to the protective effects.

  4. Enhancement of protective efficacy through adenoviral vectored vaccine priming and protein boosting strategy encoding triosephosphate isomerase (SjTPI) against Schistosoma japonicum in mice.

    Science.gov (United States)

    Dai, Yang; Wang, Xiaoting; Tang, Jianxia; Zhao, Song; Xing, Yuntian; Dai, Jianrong; Jin, Xiaolin; Zhu, Yinchang

    2015-01-01

    Schistosomiasis japonica is a zoonotic parasitic disease; developing transmission blocking veterinary vaccines are urgently needed for the prevention and control of schistosomiasis in China. Heterologous prime-boost strategy, a novel vaccination approach, is more effective in enhancing vaccine efficacy against multiple pathogens. In the present study, we established a novel heterologous prime-boost vaccination strategy, the rAdV-SjTPI.opt intramuscular priming and rSjTPI subcutaneous boosting strategy, and evaluated its protective efficacy against Schistosoma japonicum in mice. Adenoviral vectored vaccine (rAdV-SjTPI.opt) and recombinant protein vaccine (rSjTPI) were prepared and used in different combinations as vaccines in a mouse model. The specific immune responses and protective efficacies were evaluated. Furthermore, the longevity of protective efficacy was also determined. Results showed that the rAdV-SjTPI.opt priming-rSjTPI boosting strategy elicited higher levels of specific IgG responses and broad-spectrum specific cellular immune responses. The protective efficacy could reach up to nearly 70% and 50% of protection could be observed at 10 weeks after the last immunization in mice. The rAdV-SjTPI.opt intramuscular priming-rSjTPI subcutaneous boosting vaccination strategy is a novel, highly efficient, and stable approach to developing vaccines against Schistosoma japonicum infections in China.

  5. Enhancement of protective efficacy through adenoviral vectored vaccine priming and protein boosting strategy encoding triosephosphate isomerase (SjTPI against Schistosoma japonicum in mice.

    Directory of Open Access Journals (Sweden)

    Yang Dai

    Full Text Available Schistosomiasis japonica is a zoonotic parasitic disease; developing transmission blocking veterinary vaccines are urgently needed for the prevention and control of schistosomiasis in China. Heterologous prime-boost strategy, a novel vaccination approach, is more effective in enhancing vaccine efficacy against multiple pathogens. In the present study, we established a novel heterologous prime-boost vaccination strategy, the rAdV-SjTPI.opt intramuscular priming and rSjTPI subcutaneous boosting strategy, and evaluated its protective efficacy against Schistosoma japonicum in mice.Adenoviral vectored vaccine (rAdV-SjTPI.opt and recombinant protein vaccine (rSjTPI were prepared and used in different combinations as vaccines in a mouse model. The specific immune responses and protective efficacies were evaluated. Furthermore, the longevity of protective efficacy was also determined. Results showed that the rAdV-SjTPI.opt priming-rSjTPI boosting strategy elicited higher levels of specific IgG responses and broad-spectrum specific cellular immune responses. The protective efficacy could reach up to nearly 70% and 50% of protection could be observed at 10 weeks after the last immunization in mice.The rAdV-SjTPI.opt intramuscular priming-rSjTPI subcutaneous boosting vaccination strategy is a novel, highly efficient, and stable approach to developing vaccines against Schistosoma japonicum infections in China.

  6. Elucidation of the zinc-dependent regulation in Amycolatopsis japonicum enabled the identification of the ethylenediamine-disuccinate ([S,S]-EDDS) genes.

    Science.gov (United States)

    Spohn, Marius; Wohlleben, Wolfgang; Stegmann, Evi

    2016-04-01

    The actinomycete Amycolatopsis japonicum produces the complexing agent ethylenediamine-disuccinate ([S,S]-EDDS), which is an isomer of the widely industrially applied ethylenediamine-tetraacetate (EDTA). In contrast to EDTA, [S,S]-EDDS is readily biodegradable and is therefore an alternative with a favourable environmental profile. Biotechnological production of [S,S]-EDDS, however, is not currently possible because its biosynthesis is inhibited by low-micromolar zinc concentrations. Here we illustrate the development of a new strategy for identifying a biosynthetic pathway that is based on the elucidation of transcriptional regulation and the screening for binding sites of the respective regulator that controls the [S,S]-EDDS biosynthesis genes. To achieve this, we identified the zinc uptake regulator Zur in A. japonicum and showed that it mediates the repression of the zinc uptake system ZnuABCAj . The Zur-binding motif, recognized by the zinc-bound Zur protein in the upstream region of znuABCAj , was used to screen the genome, leading to the identification of the aes genes. Transcriptional analysis and shift assays reveal specific zinc-responsive regulation of the aes genes by Zur, and gene inactivation shows their involvement in [S,S]-EDDS biosynthesis. Zur-mediated zinc repression of the [S,S]-EDDS biosynthesis genes is abolished in a Δzur mutant, which offers now the opportunity to develop a biotechnological process. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  7. LC-MS/MS determination and pharmacokinetic study of seven flavonoids in rat plasma after oral administration of Cirsium japonicum DC. extract.

    Science.gov (United States)

    Zhang, Zhiyong; Jia, Peipei; Zhang, Xiaoxu; Zhang, Qiaoyue; Yang, Haotian; Shi, He; Zhang, Lantong

    2014-12-02

    Cirsium japonicum DC., a Traditional Chinese Medicine, has the curative effect of antihemorrhagic and antitumor. Pharmacological studies prove that the curative effect may relate to the flavonoids. A simple and rapid resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was first developed and validated for the quantification of seven flavonoids including pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and apigenin in rat plasma after oral administration of Cirsium japonicum DC. extract. The chromatographic separation was achieved on a C18 column with gradient elution by using a mixture of 0.1% formic acid aqueous solution and methanol as the mobile phase at a flow rate of 0.8mL/min. A tandem mass spectrometric detection was conducted using multiple-reaction monitoring (MRM) via an electrospray ionization (ESI) source in positive and negative ionization mode simultaneously. Samples were pre-treated by a single-step protein precipitation with methanol, and sulfamethoxazole was used as internal standard (IS). The optimized mass transition ion-pairs (m/z) for quantization were 623.4/315.2 for pectolinarin, 593.3/285.1 for linarin, 315.3/300.2 for pectolinarigenin, 301.2/286.2 for hispidulin, 301.2/258.2 for diosmetin, 283.0/267.9 for acacetin, 269.0/117.0 for apigenin and 252.2/155.8 for IS. After oral administration of 6mL/kg Cirsium japonicum DC. extract in rats, the maximum plasma concentration (Cmax) of pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and apigenin were 876.77±97.34ng/mL, 86.79±1.70ng/mL, 6.13±0.12ng/mL, 32.85±2.50ng/mL, 37.2±2.04ng/mL, 19.02±1.29ng/mL and 148.26±20.63ng/mL, respectively. The time to reach the maximum plasma concentration (Tmax) was 5min for pectolinarin, linarin, pectolinarigenin, hispidulin, diosmetin, acacetin and 360min for apigenin. The intra-day and inter-day precisions (RSD%) for seven compounds were less than 13.16% and 7.77% and the accuracy (RE

  8. Análisis de las alteraciones estructurales y/o regulatorias en los genes de nodulación y fijación de nitrógeno, en aislados de Bradyrhizobium japonicum que difieren en su capacidad de fijar nitrógeno

    OpenAIRE

    López, Silvina M. Y.

    2016-01-01

    La fijación biológica de nitrógeno (N) que consiste en la reducción de N2 atmosférico a NH3 realizada por algunos organismos procariotas, constituye el mayor aporte de nitrógeno a la biosfera. Entre estos organismos, los que fijan el nitrógeno asociándose simbióticamente con las plantas de la familia de las leguminosas son los que realizan el mayor aporte. Las interacciones bacteria-planta conducen a la formación de nódulos como parte de un proceso complejo, de numerosos intercambios de señal...

  9. Inoculation of Acacia mangium with Alginate Beads Containing Selected Bradyrhizobium Strains under Field Conditions: Long-Term Effect on Plant Growth and Persistence of the Introduced Strains in Soil.

    Science.gov (United States)

    Galiana, A; Prin, Y; Mallet, B; Gnahoua, G M; Poitel, M; Diem, H G

    1994-11-01

    The growth response of Acacia mangium Willd. to inoculation with selected Bradyrhizobium strains was investigated in two field trials in the Ivory Coast (West Africa). In the first trial (Anguededou), four provenances (i.e., trees originating from seeds harvested in different geographical areas) of A. mangium were inoculated with four Bradyrhizobium strains from different origins. Six months after being transplanted in the field, the heights of all inoculated trees showed a statistically significant increase of 9 to 26% compared with those of uninoculated trees, with the most effective strain being Aust 13c. After 19 months, the positive effect of inoculation on tree growth was confirmed. The effect of A. mangium provenance on tree growth was also highly significant. Trees from the Oriomo provenance of Papua New Guinea had a mean height that was 25% greater than those of other provenances. Analysis of variance showed a highly significant effect of interaction between strain and host provenance factors. Thus, most effective strain x provenance combinations could be proposed. Immunological identification of strains clearly showed that 90 to 100% of nodules from trees inoculated with three of the four Bradyrhizobium strains or from uninoculated trees contained exclusively Aust 13c 23 months after tree transplantation. This predominance of Aust 13c in nodules was still observed 42 months after tree transplantation. The second experiment (Port-Bouët), performed with a different soil, confirmed the long-term positive effect of Aust 13c on plant growth, its high competitive ability against indigenous strains, and its persistence in soil. Strain Aust 13c should thus be of great interest for inoculating A. mangium under a wide range of field conditions.

  10. Ecological Model to Predict Potential Habitats of Oncomelania hupensis, the Intermediate Host of Schistosoma japonicum in the Mountainous Regions, China.

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    Hong-Ru Zhu

    Full Text Available Schistosomiasis japonica is a parasitic disease that remains endemic in seven provinces in the People's Republic of China (P.R. China. One of the most important measures in the process of schistosomiasis elimination in P.R. China is control of Oncomelania hupensis, the unique intermediate host snail of Schistosoma japonicum. Compared with plains/swamp and lake regions, the hilly/mountainous regions of schistosomiasis endemic areas are more complicated, which makes the snail survey difficult to conduct precisely and efficiently. There is a pressing call to identify the snail habitats of mountainous regions in an efficient and cost-effective manner.Twelve out of 56 administrative villages distributed with O. hupensis in Eryuan, Yunnan Province, were randomly selected to set up the ecological model. Thirty out of the rest of 78 villages (villages selected for building model were excluded from the villages for validation in Eryuan and 30 out of 89 villages in Midu, Yunnan Province were selected via a chessboard method for model validation, respectively. Nine-year-average Normalized Difference Vegetation Index (NDVI and Land Surface Temperature (LST as well as Digital Elevation Model (DEM covering Eryuan and Midu were extracted from MODIS and ASTER satellite images, respectively. Slope, elevation and the distance from every village to its nearest stream were derived from DEM. Suitable survival environment conditions for snails were defined by comparing historical snail presence data and remote sensing derived images. According to the suitable conditions for snails, environment factors, i.e. NDVI, LST, elevation, slope and the distance from every village to its nearest stream, were integrated into an ecological niche model to predict O. hupensis potential habitats in Eryuan and Midu. The evaluation of the model was assessed by comparing the model prediction and field investigation. Then, the consistency rate of model validation was calculated in

  11. Specific anti-glycan antibodies are sustained during and after parasite clearance in Schistosoma japonicum-infected rhesus macaques.

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    Y Y Michelle Yang

    2017-02-01

    Full Text Available Human immunity to Schistosoma infection requires many years of exposure, and multiple infections and treatments to develop. Unlike humans, rhesus macaques clear an established schistosome infection naturally at the same time acquiring immunity towards re-infection. In macaques, schistosome egg production decreases after 8 weeks post-infection and by week 22, physiological impairment of the worm caused by unclarified antibody-mediated processes is observed. Since strong antibody responses have been observed against schistosome glycan antigens in human and animal infections, we here investigate if anti-glycan antibodies are associated with immunity against schistosome infections in macaques.We used a microarray containing a large repertoire of glycoprotein- and glycolipid-derived glycans from different schistosome life stages to analyse anti-glycan serum IgG and IgM from S. japonicum-infected macaques during the course of infection and self-cure. We also used an in vitro schistosomula assay to investigate whether macaque sera containing anti-glycan antibodies can kill schistosomula.Antibody responses towards schistosome glycans at week 4 post-infection were dominated by IgM while IgG was high at week 8. The profound increase in IgG was observed mainly for antibodies towards a large subset of glycans that contain (multi-fucosylated terminal GalNAcβ1-4GlcNAc (LDN, and Galβ1-4(Fucα1-3GlcNAc (LeX motifs. In general, glycans with a higher degree of fucosylation gave rise to stronger antibody responses than non-fucosylated glycans. Interestingly, even though many IgG and IgM responses had declined by week 22 post-infection, IgG towards O-glycans with highly fucosylated LDN motifs remained. When incubating macaque serum with schistosomula in vitro, schistosomula death was positively correlated with the duration of infection of macaques; macaque serum taken 22 weeks post-infection caused most schistosomula to die, suggesting the presence of potentially

  12. Effect of the Application of Rice Straw and (NH_4)_2SO_4 to a Paddy Field on the Competitive Nodulation Ability of Indigenous Bradyrhizobium Strains

    OpenAIRE

    OZAWA, Takashi; MATSUI, Eiko; KIMURA, Yoshihito

    2000-01-01

    Competitive nodulation abilities of indigenous Bradyrhizobium strains in a paddy rice field at Osaska Prefectural Agricultural and Forestry Research Center were estimated by measuring both their nodule occupancy after inoculation of the soil suspensions to soybean and N-phenyl-1-naphthylamine (NPN) uptake activity of the isolates from the soils. Three plots (50 m^2 each) in the field had been applied annually for 18 years with (NH_4)_2SO_4 and rice straw at the following rate. Plot I : 0 and ...

  13. CARACTERIZACIÓN QUÍMICA DEL GRANO MOLIDO Y LA TORTA DE SOYA DESGRASADA (Glycine max, Merr; VAR. INCA SOY 24) QUE SE EMPLEAN EN EL MEDIO DE CULTIVO PARA Bradyrhizobium elkanii

    OpenAIRE

    Aida T. Rodríguez; María C. Nápoles; M. A. Ramírez; Yamilet Gutiérrez; Daimy Costales

    2004-01-01

    Algunos derivados de la soya se han utilizado como medio de cultivo para Bradyrhizobium, demostrándose su efecto positivo sobre la dinámica de crecimiento de estos microorganismos, así como su poder inductor sobre la expresión de los genes de nodulación. El presente trabajo se llevó a cabo en el Departamento de Fisiología y Bioquímica Vegetal, perteneciente al Instituto Nacional de Ciencias Agrícolas (INCA), donde se realizó la caracterización química del grano molido y la torta desgrasada de...

  14. Combined IL-12 Plasmid and Recombinant SjGST Enhance the Protective and Anti-pathology Effect of SjGST DNA Vaccine Against Schistosoma japonicum.

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    Po-Ching Cheng

    2016-02-01

    Full Text Available Schistosomiasis is listed as one of most important tropical diseases and more than 200 million people are estimated to be infected. Development of a vaccine is thought to be the most effective way to control this disease. Recombinant 26-kDa glutathione S-transferase (rSjGST has previously been reported to achieve a worm reduction rate of 42-44%. To improve the efficiency of the vaccine against Schistosoma japonicum, we immunized mice with a combination of pcDNA vector-encoded 26-kDa SjGST (pcDNA/SjGST, IL-12 expressing-plasmid (pIL-12, and rSjGST. Co-vaccination with pcDNA/SjGST, pIL-12, and rSjGST led to a reduction in worm burden, hepatic egg burden, and the size of liver tissue granulomas than that in the untreated infection controls. In addition, we detected high levels of specific IgG, IgG1, and IgG2a against the rSjGST antigen in infected mice vaccinated with this combination of pcDNA/SjGST, pIL-12, and rSjGST. Moreover, high expression levels of Th2 cytokines, including IL-4 and IL-10, were also detected in this group, without diminished levels of IL-12, INF-γ, and TNF-α cytokines that are related to parasite killing. In conclusion, we have developed a new vaccination regimen against S. japonicum infection and shown that co-immunization with pcDNA/SjGST vaccine, pIL-12, and rSjGST has significant anti-parasite, anti-hepatic egg and anti-pathology effects in mice. The efficacy of this vaccination method should be further validated in large animals such as water buffalo. This method may help to reduce the transmission of zoonotic schistosomiasis japonica.

  15. Recombinant Sj16 from Schistosoma japonicum contains a functional N-terminal nuclear localization signal necessary for nuclear translocation in dendritic cells and interleukin-10 production.

    Science.gov (United States)

    Sun, Xi; Yang, Fan; Shen, Jia; Liu, Zhen; Liang, Jinyi; Zheng, Huanqin; Fung, Mingchiu; Wu, Zhongdao

    2016-12-01

    Sj16 is a Schistosoma japonicum-derived protein (16 kDa in molecular weight) that has been identified as an immune modulation molecule, but the mechanisms of modulation of immune responses are not known. In this report, we aimed to investigate the host immune regulation mechanism by recombinant Sj16 (rSj16) and thus illuminate the molecular mechanism of immune evasion by S. japonicum. The effect of rSj16 and rSj16 mutants on the biology of dendritic cells (DCs) was assessed by examining DC maturation, cytokine production, and expression of surface markers by flow cytometry and enzyme-linked immunosorbent assay. We found that rSj16 significantly stimulated interleukin (IL)-10 production and inhibited LPS-induced bone marrow-derived dendrite cell (BMDC) maturation in a dose-dependent manner. By using antibody neutralization experiments and IL-10-deficient (knockout) mice, we confirmed that the inhibitory effect of rSj16 on LPS-induced BMDCs is due to its induction of IL-10 production. To understand how rSj16 induces the production of IL-10, we analyzed the protein sequence and revealed two potential nuclear localization signals (NLS) in Sj16. The N-terminal NLS (NLS1) is both necessary and sufficient for translocation of rSj16 to the nucleus of BMDCs and is important for subsequent induction of IL-10 production and the inhibition of BMDC maturation by rSj16. The results of our study concluded that the ability of rSj16 to inhibit DC functions is IL-10 dependent which is operated by IL-10R signal pathway. This study also confirmed that NLS is an important domain associated with increased production of IL-10. Our findings will extend the current understanding on host-schistosome relationship and provide insight about bottleneck of parasitic control.

  16. Combined IL-12 Plasmid and Recombinant SjGST Enhance the Protective and Anti-pathology Effect of SjGST DNA Vaccine Against Schistosoma japonicum

    Science.gov (United States)

    Cheng, Po-Ching; Lin, Ching-Nan; Peng, Shih-Yi; Kang, Tsung-Fu; Lee, Kin-Mu

    2016-01-01

    Schistosomiasis is listed as one of most important tropical diseases and more than 200 million people are estimated to be infected. Development of a vaccine is thought to be the most effective way to control this disease. Recombinant 26-kDa glutathione S-transferase (rSjGST) has previously been reported to achieve a worm reduction rate of 42–44%. To improve the efficiency of the vaccine against Schistosoma japonicum, we immunized mice with a combination of pcDNA vector-encoded 26-kDa SjGST (pcDNA/SjGST), IL-12 expressing-plasmid (pIL-12), and rSjGST. Co-vaccination with pcDNA/SjGST, pIL-12, and rSjGST led to a reduction in worm burden, hepatic egg burden, and the size of liver tissue granulomas than that in the untreated infection controls. In addition, we detected high levels of specific IgG, IgG1, and IgG2a against the rSjGST antigen in infected mice vaccinated with this combination of pcDNA/SjGST, pIL-12, and rSjGST. Moreover, high expression levels of Th2 cytokines, including IL-4 and IL-10, were also detected in this group, without diminished levels of IL-12, INF-γ, and TNF-α cytokines that are related to parasite killing. In conclusion, we have developed a new vaccination regimen against S. japonicum infection and shown that co-immunization with pcDNA/SjGST vaccine, pIL-12, and rSjGST has significant anti-parasite, anti-hepatic egg and anti-pathology effects in mice. The efficacy of this vaccination method should be further validated in large animals such as water buffalo. This method may help to reduce the transmission of zoonotic schistosomiasis japonica. PMID:26891172

  17. Evaluating effect of biofertilizer on nodulation and soybean (Glycine max L plants growth characteristics under water deficit stress of seed

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    M. Tajik Khaveh

    2016-05-01

    Full Text Available In order to evaluate the effects of biofertilizer on soybean (Glycine max L. seed vigor that produced under water deficit condition and related traits, an experiment was conducted in a factorial layout based of complete randomized block design with four replications at the research greenhouse of Aboureihan campus- Tehran University, Iran. Experimental treatments were include biofertilizer (seed inoculation with Bradyrhizobium japonicum, co-inoculation with Bradyrhizobium japonicum and Pseudomonas fluorescens, co-inoculation with Bradyrhizobium japonicum and Glomus mosseae, Cultivar (Zalta Zalha and Clark×Hobbit line and water deficit stress [irrigation plants after 50 (normal irrigation, 100 (medium stress, 150 (sever stress mm evaporation from pan class A, in parents field]. Results showed that the water deficit stress had negative effects on seed quality and seedling emergence percentage, mean daily seedling emergence, root, leaf and shoot dry weight, number of nodule were decreased. ZaltaZalha cultivar had higher shoot dry weight and number of leaf compared with other cultivars. Applications of biofertilzer was effective on stem diameter, root, leaf and shoot dry weight, number of leaf and nodule and those attributes increased by co-inoculation of Bradyrhizobium japonicum and Glomus mosseae. Also, use of biofertilizer in stress levels was effective on stem dry weight. Stem dry weight was increased by Co-inoculation of cultivar seeds with Bradyrhizobium japonicum and Glomus mosseae.

  18. Symbiotic efficiency and genetic characteristics of Bradyrhizobium sp. strain UFSM LA 1.3 isolated from Lupinus albescens (H. et Arn Eficiência simbiótica e características genéticas da estirpe UFSM LA 1.3 de Bradyrhizobium sp. isolado de Lupinus albescens (H. et Arn

    Directory of Open Access Journals (Sweden)

    Marcos Roberto Dobler Stroschein

    2010-12-01

    Full Text Available Legume species belonging to the genus Lupinus are annual herb plants. The majority of them are indigenous to the Americas. They are known for nitrogen-fixing symbioses with soil bacteria collectively called rhizobia. The aim of this study was to characterize a rhizobium strain isolated from Lupinus albescens using phenotypic, symbiotic and molecular approaches. Strain UFSM LA 1.3 was tested in vitro according to several parameters: colony size, color and growing rate; acid or alkaline reaction in yeast mannitol media supplemented with bromothymol blue; gum production. Molecular characterization was evaluated by PCR technique using primers BOX A1-R and sequence analysis of the 16S-23S rDNA intergenic region (ITS. ITS sequencing fragments showed genetic similarity with Bradyrhizobium sp. The polymorphism observed by BOX-PCR have shown that strain differs from the reference strain SEMIA 928 and SEMIA 938. The symbiotic efficiency under axenic conditions of UFSM LA 1.3 was 94.6%, without statistical differences compared to the mineral nitrogen fertilized control, to which was applied solution of 400 mg of ammonium nitrate.Espécies de leguminosas pertencentes ao gênero Lupinus são plantas herbáceas anuais. A maioria é originária das Américas. Estas plantas estabelecem simbioses com bactérias do solo que realizam fixação biológica de nitrogênio coletivamente chamada de rizóbios. Caracterizou-se uma estirpe isolada de Lupinus albescens por meio de características fenotípicas, simbióticas e moleculares. A estirpe UFSM LA 1.3 foi testada in vitro de acordo com os parâmetros: tamanho de colônia; cor e taxa de crescimento; reação ácida ou básica em meio levedura manitol suplementado com azul de bromotimol; produção de goma. A caracterização molecular foi feita pela técnica de PCR usando os oligonucleotídeos BOX A1-R e seqüenciamento da região ITS. A análise da seqüência dos fragmentos da região intergênica (ITS 16S-26S r

  19. The South-to-North Water Diversion Project: effect of the water diversion pattern on transmission of Oncomelania hupensis, the intermediate host of Schistosoma japonicum in China

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    Liang You-Sheng

    2012-03-01

    Full Text Available Abstract Background The South-to-North Water Diversion Project (SNWDP is the largest national water conservancy project in China. However, the Eastern Route Project (ERP of SNWDP will refer to the habitats of Oncomelania hupensis, the intermediate host of Schistosoma japonicum. The present study was aimed at investigating the effects of some factors relating to the water diversion pattern on the spread north of O. hupensis and transmission of S. japonicum. Methods Marked snails were attached to the floating debris, and then placed on the water surface, the passage of snails through water pumps was observed. Some marked living adult snails were placed under water in the 5 spots, 15, 30, 60, 90 and 120 days later, their survival and transfer under water were investigated. 2, 4, 8, 16, 32, 64 and 128 juvenile snails, with a male: female ratio of about 1, were caged, 1 year later, their reproductions were calculated. Results The snails attached on the floating debris at 100-, 50- and 20-cm-distance from the inlet pipe of the big pump (with a diameter of 80 cm, could be absorbed into the pumps, with passing rates of 2.45%, 3.93% and 43.46%, respectively, compared with 72.07% and 91.00% for the snails at 20 cm and 10 cm-distance from the inlet pipe of the small pump (with a diameter of 20 cm. A total of 36,600 marked living snails were put into 5 ponds and ditches, with the water depths of 1-1.6 m, 15-120 days later, no marked ones were found along the ponds and ditches or in the straw packages. The juvenile snails did not reproduce until their density reached up to 8 snails (ratio of male: female of 1/0.16 m2. Conclusions During the construction of ERP of SNWDP, the risk of northward spread of schistosomiasis japonica will be decreased or eliminated as long as long-term reliable interventions for snail control are implemented.

  20. [Surveillance and forecast system of schistosomiasis in Jiangsu Province. VI. Detection technology of water infectivity based on enrichment of Schistosoma japonicum cercariae on water surface].

    Science.gov (United States)

    Qu, Guo-li; Dai, Jian-rong; Xing, Yuan-tian; Wang, Wei; Yang, Zhen-kun; Zhao, Zheng-yang; Guo, Na; Sun, Le-ping; Liang, You-Sheng

    2014-10-01

    To explore the enrichment technique of Schistosoma japonicum cercariae on the water surface, so as to establish a new method combined with the existing technology to detect the cercarial infested water body quickly and sensitively. Soybean oil, gasoline, kerosene and isophorone were screened as expanding agents. The cercariae were enriched by the thrust of the expanding agents when diffusing on the water surface, and PE adsorption film and C-6 film were applied to seize them so as to determine the infectivity of the water quickly. The relationship between the dose of expanding agents and diffusion radius were explored. Gasoline, kerosene and isophorone were suitable expanding agents, and the diffusion effect of isophorone was the best. After the enrichment by the expanding agents, the detection rate of cercariae of the method seizing cercariae with the film significantly improved in the water. This new method could effectively improve the detection rate of the cercarial infested water and is suitable for the low-degree infested water.

  1. Novel T-cell epitopes on Schistosoma japonicum SjP40 protein and their preventive effect on allergic asthma in mice.

    Science.gov (United States)

    Ren, Jiling; Hu, Lizhi; Yang, Jing; Yang, Liang; Gao, Fei; Lu, Ping; Fan, Mengyu; Zhu, Yunjuan; Liu, Junyan; Chen, Lingling; Gupta, Shimpy; Yang, Xi; Liu, Peimei

    2016-05-01

    Allergic asthma is a chronic inflammatory disease mediated by Th2 cell immune responses. Currently, immunotherapies based on immune deviation are attractive, preventive, and therapeutic strategies for asthma. Many studies have shown that intracellular bacterial infections such as mycobacteria and their components can suppress asthmatic reactions by enhancing Th1 responses, while helminth infections and their proteins can inhibit allergic asthma via immune regulation. However, some helminth proteins such as SmP40, the major egg antigen of Schistosoma mansoni, are found as Th1 type antigens. Using a panel of overlapping peptides, we identified T-cell epitopes on SjP40 protein of Schistosoma japonicum, which can induce Th1 cytokine and inhibit the production of Th2 cytokines and airway inflammation in a mouse model of allergic asthma. These results reveal a novel form of immune protective mechanism, which may play an important role in the modulating effect of helminth infection on allergic asthmatic reactions. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Low Transmission to Elimination: Rural Development as a Key Determinant of the End-Game Dynamics of Schistosoma japonicum in China

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    Robert Spear

    2017-08-01

    Full Text Available Rural development has been a critical component of China’s economic miracle since the start of economic reform in the early 1980s, both benefiting from and contributing to the nation’s rapid economic growth. This development has yielded substantial improvements of public health relevance, including contributing to major reductions in schistosomiasis prevalence. The history of schistosomiasis elimination in Japan suggests that development played a dominant causal role in that nation. We argue that it is highly probable that a similar story is playing out in at least some large regions of China. In particular, we summarize evidence from Sichuan Province which supports the case that economic development has led to improvements in rural irrigation and water supply which, together with changes in crop selection and agricultural mechanization, have all contributed to sustainable reductions in the prevalence of Schistosoma japonicum. The two major factors that have experienced major reductions are the area of snail habitat and the degree of human exposure, both through a variety of mechanisms which differ by region and economic circumstance. However, hotspots of transmission remain. Overall, however, economic development in traditionally endemic areas has provided the resources to carry out projects that have had major beneficial impacts on disease transmission that are likely to be sustainable.

  3. Impact of changing water levels and weather on Oncomelania hupensis hupensis populations, the snail host of Schistosoma japonicum, downstream of the Three Gorges Dam.

    Science.gov (United States)

    Seto, Edmund Y W; Wu, Weiping; Liu, Hong-Yun; Chen, Hong-Gen; Hubbard, Alan; Holt, Ashley; Davis, George M

    2008-06-01

    Increasing evidence indicates that dams impact riverine ecosystems and human diseases. Poyang Lake, one of the largest schistosomiasis endemic environments in China, will change due to the construction of the Yangtze River Three Gorges Dam. We assess changes in Oncomelania hupensis hupensis, the snail host for Schistosoma japonicum, in response to changing water levels and weather from 1998 to 2002. In the 5 years following the major flooding of Poyang Lake in 1998, seasonal water levels have gradually decreased, concomitant with decreases in mean and variance of fall snail densities. Nonlinear relationships suggest that the highest spring density is associated with current, 2-, and 3-month prior temperatures of 18 degrees, 9.1 degrees, and 5.8 degrees C, while the highest fall density is associated with 2- and 3-month prior water levels of 17 and 18 m, respectively. This suggests that lower, more stable water levels downstream of the dam may result in a reduction in mean fall densities and their variance. However, additional data are needed to determine whether snail populations that are typically destroyed by seasonal floods may live longer in more stable environments created by the dam.

  4. [Effects of soluble adult worm antigen and soluble egg antigen of Schistosoma japonicum on differentiation of CD4+ T cells of mice].

    Science.gov (United States)

    Yang, Xiao-Wei; Zhang, Cui; Dong, Xiao-Xiao; Li, Yong; Xu, Zhi-Peng; Zhang, Wei-Wei; Kong, Wen-Jun; Xue, Xue; Chen, Xiao-Jun; Zhu, Ji-Feng; Zhou, Sha; He, Lei; Liu, Feng; Su, Chuan

    2013-04-01

    To investigate and compare the different effects of soluble adult worm antigen (SWA) and soluble egg antigen (SEA) of Schistosoma japonicum on the differentiation of the splenocytes and CD4+ T cells of mice. The splenocytes and CD4+ T cells were prepared from the spleens of mice immunized with SWA or SEA, or the splenocytes of normal mice were harvested and stimulated with SWA or SEA in vitro. Then, the proportions of IFN-gamma and IL-4-producing cells in splenocytes, and the proportions of Th1 and Th2 cells in CD4+ T cells were determined by FACS, respectively. Compared to the SWA stimulation group, the higher proportions of IL-4-producing cells in splenocytes and of Th1 cells in CD4+ T cells were observed under the SEA stimulation group (P < 0.05). Whereas SWA induced the significantly higher proportions of IFN-gamma producing cells in splenocytes and of Th2 cells in CD4+ T cells than those in the SEA stimulation group (P < 0.05). The significantly higher levels of Th1 cells are only observed under SWA induction, however, the differentiation of Th2 cells in response to SEA stimulation is significantly more than that in response to SWA stimulation.

  5. Schistosoma japonicum risk in Jiangsu province, People’s Republic of China: identification of a spatio-temporal risk pattern along the Yangtze River

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    Kun Yang

    2013-11-01

    Full Text Available The risk for Schistosoma japonicum infection in Jiangsu province, People’s Republic of China, was investigated by a mouse bioassay. Various investigations were conducted in the period 2009-2011 with the presentation here representing the summary of the results from 45-50 sites in the marshlands along the Yangtze River’s course through the province. Indices representing three aspects of the infection were collected to assess risk: (i the proportion of sentinel points where at least one mouse infection was recorded; (ii the proportion of infected mice at each of these sites; and (iii the average worm burdens. Directional distribution analysis and scan statistics were used to explore the spatio-temporal risk pattern. The spatial distribution was oriented along the Yangtze River and the directional distributions for the proportion of infected mice and mean worm burdens were similar for the positive sentinel sites. Four statistically significant clusters were detected in 2009, but only one in 2010 and 2011, respectively. Temporal windows for infection risk were seen in June and September. The study illustrates the utility of spatio-temporal analysis in assessing the risk for schistosomiasis. This approach should be useful with respect to surveillance and response that can be expected to be increasingly applied when moving from morbidity control to transmission control.

  6. Eficiência e competitividade de variantes espontâneos isolados de estirpes de Bradyrhizobium spp recomendadas para a cultura da soja (Glycine max Effectiveness and competitiveness of spontaneous mutants isolated from Bradyrhizobium spp strains recommended for soybean crop (Glycine max

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    Fabíola Gomes de Carvalho

    2005-12-01

    Full Text Available O cultivo sucessivo de soja inoculada numa mesma área proporcionou a adaptação de uma população de rizóbios, que podem não ser tão eficientes quanto à capacidade de fixação de N2, mas apresentam alta competitividade, dificultando a introdução de novas estirpes mais eficientes. Com a finalidade de avaliar o desempenho simbiótico (eficiência e competitividade de variantes espontâneos isolados de estirpes de B. japonicum (SEMIA 5079 e SEMIA 5080 e B. elkanii (SEMIA 587 e SEMIA 5019, realizou-se um experimento em casa de vegetação onde os variantes foram inoculados isoladamente e em diferentes combinações entre os variantes e uma estirpe comprovadamente mais competitiva (SEMIA 587 ou SEMIA 5019 a partir da adição de inóculos mistos (1/1; v/v no cultivar de soja BR-16. Por meio da avaliação das variáveis analisadas (nodulação, produção de matéria de seca da parte aérea, N total acumulado na parte aérea e ocupação nodular, foi possível constatar que o determinante da maior eficiência em tratamentos co-inoculados não foi a ocupação nodular de determinada estirpe ou variante presente no inóculo, mas, sim, o tipo de interação (sinérgica ou antagônica predominante no tratamento co-inoculado e que é possível selecionar variantes eficientes e competitivos para a cultura da soja a partir de estirpes parentais que já apresentam características desejáveis para utilização em inoculantes comerciais.The continuous cultivation of inoculated soybean in the same area can determine the soil colonization with a rhizobia population presenting low nitrogen fixation effectiveness. This fact can be a problem for the establishment of a more effective population. A greenhouse experiment was carried out to evaluate the symbiotic effectiveness and competitiveness of spontaneous mutants isolated from B. japonicum (SEMIA 5079 and SEMIA 5080 and B. elkanii (SEMIA 587 and SEMIA 5019 strains. The soybean biovar BR 16 was

  7. Characterization of Schistosoma japonicum CP1412 protein as a novel member of the ribonuclease T2 molecule family with immune regulatory function.

    Science.gov (United States)

    Ke, Xue-Dan; Shen, Shuang; Song, Li-Jun; Yu, Chuan-Xin; Kikuchi, Mihoko; Hirayama, Kenji; Gao, Hong; Wang, Jie; Yin, Xuren; Yao, Yuan; Liu, Qian; Zhou, Wei

    2017-02-17

    Schistosome infection typically induces a polarized Th2 type host immune response. As egg antigen molecules play key roles in this immunoregulatory process, clarifying their functions in schistosomiasis would facilitate the development of vaccine and immunotherapeutic methods. Schistosoma japonicum (Sj) CP1412 (GenBank: AY57074.1) has been identified as a new member of the RNase T2 family with immune regulatory functions. The expression plasmid Sj CP1412-pET28a was constructed and transformed into bacteria for production of recombinant Sj CP1412 protein (rSj CP1412) via IPTG induction. The RNase activity of Sj CP1412 was predicted by bioinformatic analysis and confirmed by digesting the yeast tRNA with rSj CP1412.C57BL/6j mice were immunized with rSj CP1412, and its immune regulatory effects in vivo and in vitro were investigated. Meanwhile, the relationship between the RNase activity of Sj CP1412 and its immune regulation was observed. Sj CP1412 was confirmed as a novel RNase T2 family protein with RNase activity. Immunoblotting and RT-PCR analyses demonstrated Sj CP1412 as a protein exclusively secreted/excreted from eggs, but not cercariae and adult worms. Stimulating RAW264.7 macrophages with rSj CP1412 raised the expression of CD206, Arg-1 and IL-10, which are related to M2 type macrophage differentiation. Stimulating dendritic cells (DCs) with rSjCP1412 failed to induce their maturation, and the recombinant protein also inhibited LPS-stimulated DC maturation. Depletion of Sj CP1412 from soluble egg antigen (SEA) impaired the ability of SEA to induce M2 type polarization of RAW264.7 macrophages. Immunizing mice with rSj CP1412 induced high antibody titers, increased serum IL-4 and TGF-β levels and splenic CD4 + CD25 + Foxp3 + T cells, downregulated serum IFN-γ levels and alleviated the egg granuloma pathology of schistosome infection. In vitro stimulation by rSj CP1412 significantly increased CD4 + CD25 + Foxp3 + T cell numbers in

  8. Three-dimensional structure of Schistosoma japonicum glutathione S-transferase fused with a six-amino acid conserved neutralizing epitope of gp41 from HIV

    Science.gov (United States)

    Lim, K.; Ho, J. X.; Keeling, K.; Gilliland, G. L.; Ji, X.; Ruker, F.; Carter, D. C.

    1994-01-01

    The 3-dimensional crystal structure of glutathione S-transferase (GST) of Schistosoma japonicum (Sj) fused with a conserved neutralizing epitope on gp41 (glycoprotein, 41 kDa) of human immunodeficiency virus type 1 (HIV-1) (Muster T et al., 1993, J Virol 67:6642-6647) was determined at 2.5 A resolution. The structure of the 3-3 isozyme rat GST of the mu gene class (Ji X, Zhang P, Armstrong RN, Gilliland GL, 1992, Biochemistry 31:10169-10184) was used as a molecular replacement model. The structure consists of a 4-stranded beta-sheet and 3 alpha-helices in domain 1 and 5 alpha-helices in domain 2. The space group of the Sj GST crystal is P4(3)2(1)2, with unit cell dimensions of a = b = 94.7 A, and c = 58.1 A. The crystal has 1 GST monomer per asymmetric unit, and 2 monomers that form an active dimer are related by crystallographic 2-fold symmetry. In the binding site, the ordered structure of reduced glutathione is observed. The gp41 peptide (Glu-Leu-Asp-Lys-Trp-Ala) fused to the C-terminus of Sj GST forms a loop stabilized by symmetry-related GSTs. The Sj GST structure is compared with previously determined GST structures of mammalian gene classes mu, alpha, and pi. Conserved amino acid residues among the 4 GSTs that are important for hydrophobic and hydrophilic interactions for dimer association and glutathione binding are discussed.

  9. Xiphinema japonicum n. sp. (Nematoda: Longidorinae) from the Rhizosphere of Japanese Podocarpus macrophyllus (Thunb.), a Cryptic Species Related to Xiphinema bakeri Williams, 1961.

    Science.gov (United States)

    Zhao, Lirong; Ye, Weimin; Maria, Munawar; Pedram, Majid; Gu, Jianfeng

    2017-12-01

    Xiphinema japonicum n. sp., isolated in Ningbo, China, from the rhizosphere of Podocarpus macrophyllus (Thunb.) imported from Japan is described. The new species belongs to Xiphinema non-americanum group 7 and is characterized by medium body length (3.0-3.7 mm), total stylet length 190-201 μm, vulva located anteriorly (V = 30.5%-35.3%), two equally developed female genital branches without uterine differentiation (no Z or pseudo-Z organ and/or spines in the uteri), short tail, convex-conoid with subdigitate peg in terminus, and absence of males. The species has four juvenile developmental stages (J1 was not found). The polytomous identification codes of the new species are (codes in parentheses are exceptions) A4-B4-C4-D5(4)-E2(3)-F3(4)-G2(3)-H2-I3-J4-K?-L1. Morphologically, the new species is mainly characterized by combination of the codes C4 and E2(3), making the species unique and different from other species in the genus. It is most similar to the North American species Xiphinema bakeri, herein considered as its cryptic species by the nature of high morphological similarity, but with significant differences in DNA sequences in nearly full length 18S, ITS1, 28S D2/D3, and cytochrome c oxidase subunit 1 sequences. The second morphologically close species is Xiphinema setariae from which the new species could be separated based on morphological and molecular characters. Morphological comparisons with phylogenetically related species are also discussed.

  10. Isojacareubin from the Chinese Herb Hypericum japonicum: Potent Antibacterial and Synergistic Effects on Clinical Methicillin-Resistant Staphylococcus aureus (MRSA

    Directory of Open Access Journals (Sweden)

    Gen-Chun Wang

    2012-07-01

    Full Text Available Through bioassay-guided fractionation of the extracts from the aerial parts of the Chinese herb Hypericum japonicum Thunb. Murray, Isojacareubin (ISJ was characterized as a potent antibacterial compound against the clinical methicillin-resistant Staphylococcus aureus (MRSA. The broth microdilution assay was used to determine the minimal inhibitory concentrations (MICs and minimal bactericidal concentrations (MBCs of ISJ alone. The results showed that its MICs/MBCs ranged from 4/16 to 16/64 μg/mL, with the concentrations required to inhibit or kill 50% of the strains (MIC50/MBC50 at 8/16 μg/mL. Synergistic evaluations of this compound with four conventional antibacterial agents representing different types were performed by the chequerboard and time-kill tests. The chequerboard method showed significant synergy effects when ISJ was combined with Ceftazidime (CAZ, Levofloxacin (LEV and Ampicillin (AMP, with the values of 50% of the fractional inhibitory concentration indices (FICI50 at 0.25, 0.37 and 0.37, respectively. Combined bactericidal activities were also observed in the time-kill dynamic assay. The results showed the ability of ISJ to reduce MRSA viable counts by log10CFU/mL at 24 h of incubation at a concentration of 1 × MIC were 1.5 (LEV, additivity, 0.92 (CAZ, indifference and 0.82 (AMP, indifference, respectively. These in vitro anti-MRSA activities of ISJ alone and its synergy with conventional antibacterial agents demonstrated that ISJ enhanced their efficacy, which is of potential use for single and combinatory therapy of patients infected with MRSA.

  11. SjCRT, a recombinant Schistosoma japonicum calreticulin, induces maturation of dendritic cells and a Th1-polarized immune response in mice

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    Lizhen Ma

    2017-11-01

    Full Text Available Abstract Background It is well known that immunization of radiation-attenuated (RA schistosoma cercariae or schistosomula can induce high levels of protective immunity against schistosoma cercariae reinfection in many animals. Many studies have shown that the Th1 cellular immune response is crucial for the protective effect elicited by RA schistosomula. However, the molecular mechanism of this strong protective immunity remains unclear. Methods The expression profiles of Schistosoma japonicum calreticulin (SjCRT in RA and normal schistosoma-derived cells were investigated by flow cytometry. The effect of recombinant SjCRT (rSjCRT on mouse dendritic cells (DCs was determined by FACS, ELISA and RT-PCR analysis. We also analyzed the effects of SjCRT on the activation of spleen cells from mice immunized with rSjCRT by detecting lymphocyte proliferation and the cytokine profiles of splenocytes. Results We found that the expression level of SjCRT in the cells from RA larvae was significantly higher than that in cells from normal schistosomula at early stages of development (day 4. The results of effect of rSjCRT on mouse DCs showed that rSjCRT could induce phenotypic and functional maturation of DCs, and SjCRT bound to the surface of DCs through the CD91 receptor and could be engulfed by DCs. The results of activation of splenocytes from mice immunized with rSjCRT also demonstrate that rSjCRT can effectively stimulate the proliferative response of splenic lymphocytes, elicit splenocytes from immunized mice to secrete high levels of IFN-γ, TNF-α and IL-4, and activate CD4+ T cells to produce high levels of IFN-γ. Conclusion SjCRT is one of the immunostimulatory molecules released from RA schistosomula cells, might play a crucial role in conferring a Th1-polarized immune response induced by RA cercariae/schistosomula in mice, and is a candidate molecule responsible for the high levels of protective immunity induced by RA schistosomula.

  12. Genetic Evidence of Contemporary Dispersal of the Intermediate Snail Host of Schistosoma japonicum: Movement of an NTD Host Is Facilitated by Land Use and Landscape Connectivity.

    Directory of Open Access Journals (Sweden)

    Jennifer R Head

    2016-12-01

    Full Text Available While the dispersal of hosts and vectors-through active or passive movement-is known to facilitate the spread and re-emergence of certain infectious diseases, little is known about the movement ecology of Oncomelania spp., intermediate snail host of the parasite Schistosoma japonicum, and its consequences for the spread of schistosomiasis in East and Southeast Asia. In China, despite intense control programs aimed at preventing schistosomiasis transmission, there is evidence in recent years of re-emergence and persistence of infection in some areas, as well as an increase in the spatial extent of the snail host. A quantitative understanding of the dispersal characteristics of the intermediate host can provide new insights into the spatial dynamics of transmission, and can assist public health officials in limiting the geographic spread of infection.Oncomelania hupensis robertsoni snails (n = 833 were sampled from 29 sites in Sichuan, China, genotyped, and analyzed using Bayesian assignment to estimate the rate of recent snail migration across sites. Landscape connectivity between each site pair was estimated using the geographic distance distributions derived from nine environmental models: Euclidean, topography, incline, wetness, land use, watershed, stream use, streams and channels, and stream velocity. Among sites, 14.4% to 32.8% of sampled snails were identified as recent migrants, with 20 sites comprising >20% migrants. Migration rates were generally low between sites, but at 8 sites, over 10% of the overall host population originated from one proximal site. Greater landscape connectivity was significantly associated with increased odds of migration, with the minimum path distance (as opposed to median or first quartile emerging as the strongest predictor across all environmental models. Models accounting for land use explained the largest proportion of the variance in migration rates between sites. A greater number of irrigation channels

  13. In vitro screening of soil bacteria for inhibiting phytopathogenic fungi ...

    African Journals Online (AJOL)

    Bradyrhizobium japonicum could highly inhibit the mycelial growth of five moulds (Botrytis cinerea, Phoma medicaginis, Fusarium verticilloides, Rhizoctonia solani and Phytophtora infestans) with a growth inhibition varying between 12.38 and 37.61%. 12 Bacillus strains and five Pseudomonas strains were antagonistic to ...

  14. African Journal of Biotechnology - Vol 12, No 20 (2013)

    African Journals Online (AJOL)

    Co-inoculation effects of Bradyrhizobium japonicum and Azospirillum sp. on competitive nodulation and rhizosphere eubacterial community structures of ... Induced gene expression in wheat seedlings treated with a crude extract of Agapanthus africanus L. prior to leaf rust infection · EMAIL FREE FULL TEXT EMAIL FREE ...

  15. Dicty_cDB: AFO507 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ne) Bradyrhizobium japonicum USDA 1... 35 7.1 AF438165_8( AF438165 |pid:none) Camelpox virus M-96 from Kaz...akhsta... 34 9.3 AY698061_1( AY698061 |pid:none) Schistosoma mansoni nuclear recept

  16. Growth, nodulation and yield of black gram [ Vigna mungo (L.) Hepper

    African Journals Online (AJOL)

    EM (effective microorganisms) is a commercial biofertilizer mainly consists of photosynthetic and lactic acid bacteria, yeast and actinomycetes. The present study was undertaken to investigate the effect of EM application and two strains of nitrogen fixing Bradyrhizobium japonicum (TAL- 102 and MN-S) on plant growth, ...

  17. Vigna mungo (L.) Hepper

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-02

    Nov 2, 2009 ... EM (effective microorganisms) is a commercial biofertilizer mainly consists of photosynthetic and lactic acid bacteria, yeast and actinomycetes. The present study was undertaken to investigate the effect of. EM application and two strains of nitrogen fixing Bradyrhizobium japonicum (TAL- 102 and MN-S) on.

  18. 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.

  19. 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.

  20. Gclust Server: 19847 [Gclust Server

    Lifescience Database Archive (English)

    Full Text Available 19847 Npun1_NpF5504 Cluster Sequences Related Sequences(157) 395 blr6297 [Bradyrhiz...obium japonicum] 5 1.00e-10 0.0 0.0 8.0 0.0 9.68 0.0 Show 19847 Cluster ID 19847 Sequence ID Npun1_NpF5504 L

  1. RHIZOBIUM CROSS-INOCULATION

    African Journals Online (AJOL)

    1993-01-13

    Jan 13, 1993 ... Six introduced and one native nitrogen fixing tree species were grown in a potted non-sterilized Inceptisol inoculated with: species-specific rhizobia strains; Rhizobium strain TAL 1145, a Leucaena isolate;. Bradyrhizobium japonicum strain TAL 169, and without inoculation. Twelve weeks later, significant.

  2. Effects of biofertilizers on grain yield and protein content of two ...

    African Journals Online (AJOL)

    Biofertilizers can affect the quality and quantity of crop. In order to study the effects of biofertilizers on grain yield and protein content of ... It therefore seems that biofertilizers can be considered as a replacement for part of chemical fertilizers in soybean production. Key words: Bacillus, Bradyrhizobium japonicum, phosphate ...

  3. Diversity and efficiency of bradyrhizobium strains isolated from soil samples collected from around sesbania virgata roots using cowpea as trap species Diversidade e eficiência simbiótica de estirpes de bradyrhizobium capturadas próximo ao sistema radicular de sesbania virgata usando caupi como planta-isca

    Directory of Open Access Journals (Sweden)

    Ligiane Aparecida Florentino

    2010-08-01

    Full Text Available The genetic diversity of ten Bradyrhizobium strains was evaluated for tolerance to high temperatures, to different salinity levels and for the efficiency of symbiosis with cowpea plants (Vigna unguiculata (L. Walp.. Eight of these strains were isolated from nodules that appeared on cowpea after inoculation with suspensions of soil sampled from around the root system of Sesbania virgata (wand riverhemp in ecosystems of South Minas Gerais. The other two strains used in our analyses as references, were from the Amazon and are currently recommended as cowpea inoculants. Genetic diversity was analyzed by amplifying repetitive DNA elements with the BOX primer, revealing high genetic diversity with each strain presenting a unique band profile. Leonard jar assays showed that the strains UFLA 03-30 and UFLA 03-38 had the highest N2-fixing potentials in symbiosis with cowpea. These strains had more shoot and nodule dry matter, more shoot N accumulation, and a higher relative efficiency than the strains recommended as inoculants. All strains grew in media of pH levels ranging from 4.0 to 9.0. The strains with the highest N2-fixing efficiencies in symbiosis with cowpea were also tolerant to the greatest number of antibiotics. However, these strains also had the lowest tolerance to high salt concentrations. All strains, with the exceptions of UFLA 03-84 and UFLA 03-37, tolerated temperatures of up to 40 ºC. The genetic and phenotypic characteristics of the eight strains isolated from soils of the same region were highly variable, as well as their symbiotic efficiencies, despite their common origin. This variability highlights the importance of including these tests in the selection of cowpea inoculant strains.Dez estirpes de rizóbios, sendo oito isoladas de amostras de solos coletadas próximo ao sistema radicular de Sesbania virgata, no Sul de Minas Gerais, e duas recomendadas como inoculante para o caupi (Vigna unguiculata (L. Walp. usadas como refer

  4. Generation of a rabbit single-chain fragment variable (scFv) antibody for specific detection of Bradyrhizobium sp. DOA9 in both free-living and bacteroid forms.

    Science.gov (United States)

    Vu, Nguyen Xuan; Pruksametanan, Natcha; Srila, Witsanu; Yuttavanichakul, Watcharin; Teamtisong, Kamonluck; Teaumroong, Neung; Boonkerd, Nantakorn; Tittabutr, Panlada; Yamabhai, Montarop

    2017-01-01

    A simple and reliable method for the detection of specific nitrogen-fixing bacteria in both free-living and bacteroid forms is essential for the development and application of biofertilizer. Traditionally, a polyclonal antibody generated from an immunized rabbit was used for detection. However, the disadvantages of using a polyclonal antibody include limited supply and cross-reactivity to related bacterial strains. This is the first report on the application of phage display technology for the generation of a rabbit recombinant monoclonal antibody for specific detection and monitoring of nitrogen-fixing bacteria in both free-living form and in plant nodules. Bradyrhizobium sp. DOA9, a broad host range soil bacteria, originally isolated from the root nodules of Aeschynomene americana in Thailand was used as a model in this study. A recombinant single-chain fragment variable (scFv) antibody library was constructed from the spleen of a rabbit immunized with DOA9. After three rounds of biopanning, one specific phage-displayed scFv antibody, designated bDOA9rb8, was identified. Specific binding of this antibody was confirmed by phage enzyme-linked immunosorbent assay (phage ELISA). The phage antibody could bind specifically to DOA9 in both free-living cells (pure culture) and bacteroids inside plant nodules. In addition to phage ELISA, specific and robust immunofluorescence staining of both free-living and bacteroid forms could also be observed by confocal-immunofluorescence imaging, without cross-reactivity with other tested bradyrhizobial strains. Moreover, specific binding of free scFv to DOA9 was also demonstrated by ELISA. This recombinant antibody can also be used for the study of the molecular mechanism of plant-microbe interactions in the future.

  5. Inibição da colonização por Bradyrhizobium elkanii mas não por Glomus intraradices em soja pelo ativador de defesa vegetal BTH

    Directory of Open Access Journals (Sweden)

    Hoffmann Lúcia Vieira

    2001-01-01

    Full Text Available O produto BTH controla doenças através da ativação dos mecanismos de resistência de plantas, cujo modo de ação tem sido caracterizado através da identificação de genes ou enzimas ativados durante esta reação. Os mesmos genes e enzimas podem ser ativados como parte da reação das plantas à colonização por microssimbiontes mutualistas. Então, a aplicação de BTH pode inibir também essas simbioses. Um experimento foi conduzido para verificar se BTH tem efeito sobre a colonização de raízes de soja pelos microssimbiontes Glomus intraradices e Bradyrhizobium elkanii. Plantas controle ou inoculadas com o fungo micorrízico ou rizóbio foram tratadas com 20 mL de BTH 25 mg L-1 por meio da aplicação em sementes, via radicular ou via foliar. As plantas que receberam aplicação radicular de BTH tiveram desenvolvimento vegetativo reduzido. A colonização de raízes por B. elkanii foi menor nas plantas que receberam aplicação de BTH via radicular do que naquelas não tratadas ou tratadas por via foliar apenas. A colonização de raízes de soja por G. intraradices não foi afetada por BTH em quaisquer dos modos de aplicação. A aplicação radicular de BTH deve ser evitada na cultura da soja. Não há indício de restrição de aplicação foliar do produto.

  6. Molecular characterization of thyroid hormone receptor beta from Schistosoma japonicum and assessment of its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice

    Directory of Open Access Journals (Sweden)

    Qiu Chunhui

    2012-08-01

    Full Text Available Abstract Background Thyroid hormones (TH modulate growth, development and differentiation and metabolic processes by interacting with thyroid hormone receptors (THRs. The purpose of this study was to identify a novel thyroid hormone receptor beta encoding gene of Schistosoma japonicum (SjTHRβ and to investigate its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice. Methods The full-length cDNA sequence of SjTHRβ, its gene organization, and its transcript levels were characterized, and the phylogenetic relationship between THR, RAR and RXR from other organisms were analysis, the ability of this protein binding to a conserved DNA core motif, and its potential as a vaccine candidate antigen against schistosomiasis in BALB/c mice were evaluated. Results The SjTHRβ cDNA was cloned, verified by 5’ and 3’ Rapid Amplification of cDNA Ends and shown to be polyadenylated at the 3’end, suggesting the transcript is full-length. SjTHRβ is homologous to THRs from other species and has a predicted conservative DNA binding domain and ligand binding domain that normally characterizes these receptors. A comparative quantitative PCR analysis showed that SjTHRβ was the highest expressed in 21d worms and the lowest in 7 d and 13 d schistosomula. The cDNA corresponding to DNA binding domain (SjTHRβ-DBD and ligand binding domain (SjTHRβ-LBD were cloned and subsequently expressed in E coli. The expressed proteins were used to immunize mice and generate specific serum against recombinant SjTHRβ (rSjTHRβ. Western blotting revealed that anti-rSjTHRβ-LBD serum recognized two protein bands in extracts from 21 d worm with molecular sizes of approximately 95 kDa and 72 kDa. Electrophoretic mobility shift assay (EMSA analysis showed that rSjTHRβ-DBD could bind to a conserved DNA core motif. Immunization of BALB/c mice with rSjTHRβ-LBD could induce partial protective efficacy(27.52% worm reduction and 29.50% liver eggs

  7. Characterization of a gene family encoding SEA (sea-urchin sperm protein, enterokinase and agrin-domain proteins with lectin-like and heme-binding properties from Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Evaristus Chibunna Mbanefo

    Full Text Available BACKGROUND: We previously identified a novel gene family dispersed in the genome of Schistosoma japonicum by retrotransposon-mediated gene duplication mechanism. Although many transcripts were identified, no homolog was readily identifiable from sequence information. METHODOLOGY/PRINCIPAL FINDINGS: Here, we utilized structural homology modeling and biochemical methods to identify remote homologs, and characterized the gene products as SEA (sea-urchin sperm protein, enterokinase and agrin-domain containing proteins. A common extracellular domain in this family was structurally similar to SEA-domain. SEA-domain is primarily a structural domain, known to assist or regulate binding to glycans. Recombinant proteins from three members of this gene family specifically interacted with glycosaminoglycans with high affinity, with potential implication in ligand acquisition and immune evasion. Similar approach was used to identify a heme-binding site on the SEA-domain. The heme-binding mode showed heme molecule inserted into a hydrophobic pocket, with heme iron putatively coordinated to two histidine axial ligands. Heme-binding properties were confirmed using biochemical assays and UV-visible absorption spectroscopy, which showed high affinity heme-binding (K D = 1.605×10(-6 M and cognate spectroscopic attributes of hexa-coordinated heme iron. The native proteins were oligomers, antigenic, and are localized on adult worm teguments and gastrodermis; major host-parasite interfaces and site for heme detoxification and acquisition. CONCLUSIONS: The results suggest potential role, at least in the nucleation step of heme crystallization (hemozoin formation, and as receptors for heme uptake. Survival strategies exploited by parasites, including heme homeostasis mechanism in hemoparasites, are paramount for successful parasitism. Thus, assessing prospects for application in disease intervention is warranted.

  8. Sj-FABPc fatty-acid-binding protein of the human blood fluke Schistosoma japonicum: structural and functional characterization and unusual solvent exposure of a portal-proximal tryptophan residue.

    Science.gov (United States)

    Kennedy, M W; Scott, J C; Lo, S; Beauchamp, J; McManus, D P

    2000-07-01

    Sj-FABPc of the blood fluke of humans, Schistosoma japonicum, is a member of the FABP/P2/CRBP/CRABP family of beta-barrel cytosolic fatty-acid-binding and retinoid-binding proteins. Sj-FABPc has at least eight different variants encoded by a single-copy polymorphic gene. In fluorescence-based assays, recombinant Sj-FABPc was found to bind 11-(dansylamino)undecanoic acid (DAUDA), inducing a shift in peak fluorescence emission from 543 to 493 nm. A similar spectral change was observed in dansyl-amino-octanoic acid (in which the dansyl fluorophore is attached at the alpha-carbon rather than the omega-carbon of DAUDA), indicating that the ligand enters entirely into the binding site. Sj-FABPc also bound the naturally fluorescent cis-parinaric acid, as well as oleic acid and arachidonic acid, by competition, but not all-trans-retinol. Dissociation constants were, for cis-parinaric acid, K(d)=2.5+/-0.1 microM (mean+/-S.E.M.) and an apparent stoichiometry consistent with one binding site per molecule of Sj-FABPc and, for oleic acid, K(i) approximately 80 nM. A deletion mutant from which alpha-II was absent failed to bind ligand. Sj-FABPc modelled well to known structures of the protein family; an unusually solvent-exposed Trp side chain was evident adjacent to the presumptive portal through which ligand is thought to enter and leave. Intrinsic fluorescence analyses of Sj-FABPc and of the deletion mutant (from which Trp-27 is absent) confirmed the unusual disposition of this side chain. Virtually all members of the FABP/P2/CRBP/CRABP protein family have prominent hydrophobic side chains in this position, with the exception of liver FABP and ileal FABP, which instead have charged side chains. Liver FABP is known to be distinct from other members of the protein family in that it does not seem to contact membranes to collect and deposit its ligand. It is therefore postulated that the unusually positioned apolar side chains in Sj-FABPc and others in the family are important in

  9. Genome Sequence of the Chemolithoautotrophic Nitrite-Oxidizing Bacterium Nitrobacter winogradskyi Nb-255

    OpenAIRE

    Starkenburg, Shawn R.; Chain, Patrick S. G.; Sayavedra-Soto, Luis A.; Hauser, Loren; Land, Miriam L.; Larimer, Frank W.; Stephanie A Malfatti; Klotz, Martin G.; Bottomley, Peter J.; Arp, Daniel J.; Hickey, William J.

    2006-01-01

    The alphaproteobacterium Nitrobacter winogradskyi (ATCC 25391) is a gram-negative facultative chemolithoautotroph capable of extracting energy from the oxidation of nitrite to nitrate. Sequencing and analysis of its genome revealed a single circular chromosome of 3,402,093 bp encoding 3,143 predicted proteins. There were extensive similarities to genes in two alphaproteobacteria, Bradyrhizobium japonicum USDA110 (1,300 genes) and Rhodopseudomonas palustris CGA009 CG (815 genes). Genes encodin...

  10. H:\\PMKER 25(3)\\BEUGRE F..xps

    African Journals Online (AJOL)

    AISA

    plateau, afin d'évaluer l'incidence d'un herbicide et de l'inoculation des semences avant le semis sur la croissance du soja. La variété de soja (Glycine max), Doko, ainsi qu'un inoculum fait à partir de la souche. IRAT-FA3 de Bradyrhizobium japonicum ont été utilisés. Le Glyphosate, un herbicide total et systémique.

  11. Isoliquiritigenin, a strong nod gene- and glyceollin resistance-inducing flavonoid from soybean root exudate.

    Science.gov (United States)

    Kape, R; Parniske, M; Brandt, S; Werner, D

    1992-01-01

    Isoflavonoid signal molecules from soybean (Glycine max (L.) Merr.) seed and root exudate induce the transcription of nodulation (nod) genes in Bradyrhizobium japonicum. In this study, a new compound with symbiotic activity was isolated from soybean root exudate. The isolated 2',4',4-trihydroxychalcone (isoliquiritigenin) is characterized by its strong inducing activity for the nod genes of B. japonicum. These genes are already induced at concentrations 1 order of magnitude below those required of the previously described isoflavonoid inducers genistein and daidzein. Isoliquiritigenin is also a potent inducer of glyceollin resistance in B. japonicum, which renders this bacterium insensitive to potentially bactericidal concentrations of glyceollin, the phytoalexin of G. max. No chemotactic effect of isoliquiritigenin was observed. The highly efficient induction of nod genes and glyceollin resistance by isoliquiritigenin suggests the ecological significance of this compound, although it is not a major flavonoid constituent of the soybean root exudate in quantitative terms. PMID:1622242

  12. Ribotyping of rhizobia nodulating Acacia mangium and Paraserianthes falcataria from different geographical areas in Indonesia using PCR-RFLP-SSCP (PRS) and sequencing.

    Science.gov (United States)

    Clapp, J P; Mansur, I; Dodd, J C; Jeffries, P

    2001-04-01

    Acacia mangium and Paraserianthes falcataria are leguminous tree species widely grown for timber in Indonesia and other tropical countries, yet little is known about the identity of their rhizobial symbionts. Polymerase chain reaction-restriction fragment length polymorphism-single-strand conformational polymorphism (PRS) analysis of the 16S rRNA gene was used along with sequencing to assess the diversity of 57 rhizobia isolated from nodules of A. mangium and P. falctaria in Indonesia. In total, 26 rhizobia isolated from A. mangium were analysed by PRS and sequencing. The PRS patterns indicated that 12 (46%) clustered with Bradyrhizobium elkanii, 13 (50%) with B. lianoningense/japonicum and one (4%) with Mesorhizobium loti. Thirty-one isolates were analysed from P. falcataria: five (16%) clustered with B. elkanii and 26 (84%) with B. lianoningense/japonicum. These results were confirmed by phylogenetic analysis of sequences. Intraspecific diversity of the 16S rRNA genes from rhizobia nodulating A. mangium and P. falcataria revealed by PRS was low, only one genotype was found within the isolates that clustered with B. elkanii and two within the B. liaoningense/japonicum group. These Bradyrhizobium species are apparently ubiquitous throughout the Indonesian archipelago and it is clear why the two tree species are able to successfully establish outside their native range without the need for inoculation with indigenous rhizobia.

  13. Genetic diversity of nodulating and non-nodulating rhizobia associated with wild soybean (Glycine soja Sieb. & Zucc.) in different ecoregions of China.

    Science.gov (United States)

    Wu, Li Juan; Wang, Hai Qing; Wang, En Tao; Chen, Wen Xin; Tian, Chang Fu

    2011-06-01

    A total of 99 bacterial isolates that originated from root nodules of Glycine soja were characterized with restriction analyses of amplified 16S ribosomal DNA and 16S-23S rDNA intergenic spacers (ITS), and sequence analyses of 16S rRNA, rpoB, atpD, recA and nodC genes. When tested for nodulation of G. soja, 72 of the isolates were effective symbionts, and these belonged to five species: Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium yuanmingense, Bradyrhizobium liaoningense and Sinorhizobium fredii. All of these, except some B. yuanmingense strains, also formed effective nodules on the domesticated soybean Glycine max. The remaining 27 isolates did not nodulate either host, but were identified as Rhizobium. Phylogeny nodC in the G. soja symbionts suggested that this symbiosis gene was mainly maintained by vertical gene transfer. Different nodC sublineages and rrs-ITS clusters reflected the geographic origins of isolates in this study. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  14. Symbiotic effectiveness of acid-tolerant Bradyrhizobium strains with ...

    African Journals Online (AJOL)

    Further, symbiotic effectiveness of these strains was determined under the polyhouse conditions in sterilized soil (pH 4.5). Highest and lowest symbiotic characters, dry matter production and nitrogen improvement per plant were observed in PSR001 and NSR008 inoculated plants, respectively. All the examined isolates ...

  15. Interrelationships between Bacillus sp. CHEP5 and Bradyrhizobium ...

    Indian Academy of Sciences (India)

    2014-10-20

    Oct 20, 2014 ... rolfsii and symbiosis on peanut plants. MARÍA SOLEDAD FIGUEREDO, MARÍA LAURA TONELLI, TANIA TAURIAN, JORGE ANGELINI,. FERNANDO IBAÑEZ, LUCIO VALETTI, VANINA MUÑOZ, MARÍA SOLEDAD ANZUAY, LILIANA LUDUEÑA and ADRIANA FABRA*. Departamento de Ciencias Naturales, ...

  16. Effect of Inoculating Bradyrhizobium on Phosphorus Use Efficiency ...

    African Journals Online (AJOL)

    of P solubilizing bacteria increased P content in grain and straw by 10.72 and. 31.94%, respectively, over the ..... 1996. Root colonization of maize and lettuce by bioluminescent Rhizobium leguminosaurm biovar phaseoli. .... Isolation and characterization of soybean-associated bacteria and their potential for plant growth ...

  17. Effect of Inoculating Bradyrhizobium on Phosphorus Use Efficiency ...

    African Journals Online (AJOL)

    P use would be very useful. A two site experiment conducted at Metahara Sugar. Estate showed that P application (0 to 60 kg P ha-1) had no significant effect on sugarcane yield (Agricultural Services, 1974). This might be ascribed to the high P precipitation caused by presence of high calcium carbonate (BAI, 2009).

  18. Isolation and selection of Bradyrhizobium from the root nodules of ...

    African Journals Online (AJOL)

    Makmal Sains Rumpai

    2012-07-19

    Jul 19, 2012 ... biofertilizer for providing greater stability in production .... Nodulation test by Leonard Bottle Jar Assembly (LBJA) was conducted in the Soil ... seedlings were placed in a tray filled with water and the roots were examined for ...

  19. Interrelationships between Bacillus sp. CHEP5 and Bradyrhizobium ...

    Indian Academy of Sciences (India)

    1 Fernando Ibañez1 Lucio Valetti1 Vanina Muñoz1 María Soledad Anzuay1 Liliana Ludueña1 Adriana Fabra1. Departamento de Ciencias Naturales, Facultad de Ciencias Exactas, Físico-quimicas y Naturales, Universidad Nacional de Río ...

  20. The Rhizobium etli rpoN Locus: DNA Sequence Analysis and Phenotypical Characterization of rpoN, ptsN, and ptsA Mutants

    OpenAIRE

    Michiels, Jan; Van Soom, Tom; D’hooghe, Inge; Dombrecht, Bruno; Benhassine, Traki; de Wilde, Petra; Vanderleyden, Jos

    1998-01-01

    The rpoN region of Rhizobium etli was isolated by using the Bradyrhizobium japonicum rpoN1 gene as a probe. Nucleotide sequence analysis of a 5,600-bp DNA fragment of this region revealed the presence of four complete open reading frames (ORFs), ORF258, rpoN, ORF191, and ptsN, coding for proteins of 258, 520, 191, and 154 amino acids, respectively. The gene product of ORF258 is homologous to members of the ATP-binding cassette-type permeases. ORF191 and ptsN are homologous to conserved ORFs f...

  1. Rhizobium strains differ considerably in outer membrane permeability and polymyxin B resistance.

    Science.gov (United States)

    Komaniecka, Iwona; Zamłyńska, Katarzyna; Zan, Radosław; Staszczak, Magdalena; Pawelec, Jarosław; Seta, Irena; Choma, Adam

    2016-01-01

    Six rhizobium (Rhizobium leguminosarum bv. Trifolii TA1, Sinorhizobium meliloti 1021, Mesorhizobium huakuii IFO 15243(T), Ochrobactrum lupini LUP 21(T), Bradyrhizobium japonicum USDA110 and B. elkanii USDA 76) and two Escherichia coli strains (E. coli ATCC 25922 and E. coli HB 101) were compared in respect to polymyxin B and EDTA resistance, as well as bacterial outer membrane (OM) permeability to a fluorescent hydrophobic agent (N-phenyl-1-naphthylamine - NPN). TEM (Transmission Electron Microscopy) and a microbial test demonstrated that all the rhizobia were much more resistant to polymyxin B in comparison with E. coli strains. EDTA and polymyxin B enhance permeability of B. japonicum and O. lupini OM. Other rhizobia incorporated NPN independently of the presence of membrane-deteriorating agents; however, the level of fluorescence (measured as NPN absorption) was strain dependent.

  2. Nitrogen-fixing rhizobial strains isolated from Desmodium incanum DC in Argentina: Phylogeny, biodiversity and symbiotic ability.

    Science.gov (United States)

    Toniutti, María Antonieta; Fornasero, Laura Viviana; Albicoro, Francisco Javier; Martini, María Carla; Draghi, Walter; Alvarez, Florencia; Lagares, Antonio; Pensiero, José Francisco; Del Papa, María Florencia

    2017-07-01

    Desmodium spp. are leguminous plants belonging to the tribe Desmodieae of the subfamily Papilionoideae. They are widely distributed in temperated and subtropical regions and are used as forage plants, for biological control, and in traditional folk medicine. The genus includes pioneer species that resist the xerothermic environment and grow in arid, barren sites. Desmodium species that form nitrogen-fixing symbiosis with rhizobia play an important role in sustainable agriculture. In Argentina, 23 native species of this genus have been found, including Desmodium incanum. In this study, a total of 64 D. incanum-nodulating rhizobia were obtained from root nodules of four Argentinean plant populations. Rhizobia showed different abiotic-stress tolerances and a remarkable genetic diversity using PCR fingerprinting, with more than 30 different amplification profiles. None of the isolates were found at more than one site, thus indicating a high level of rhizobial diversity associated with D. incanum in Argentinean soils. In selected isolates, 16S rDNA sequencing and whole-cell extract MALDI TOF analysis revealed the presence of isolates related to Bradyrhizobium elkanii, Bradyrhizobium japonicum, Bradyrhizobium yuanmingense, Bradyrhizobium liaoningense, Bradyrhizobium denitrificans and Rhizobium tropici species. In addition, the nodC gene studied in the selected isolates showed different allelic variants. Isolates were phenotypically characterized by assaying their growth under different abiotic stresses. Some of the local isolates were remarkably tolerant to high temperatures, extreme pH and salinity, which are all stressors commonly found in Argentinean soils. One of the isolates showed high tolerance to temperature and extreme pH, and produced higher aerial plant dry weights compared to other inoculated treatments. These results indicated that local isolates could be efficiently used for D. incanum inoculation. Copyright © 2017 Elsevier GmbH. All rights reserved.

  3. Sur quelques aspects de la production du soja (Glycine max L. au Congo : essais préliminaires

    Directory of Open Access Journals (Sweden)

    Mandimba, GR.

    1991-01-01

    Full Text Available About some cropping systems of soybean (Glycine max. L. in Congo : first results. Field experiments were conducted to assess the response of soybean Glycine max cv. FN3 to N fertilization and inoculation respectively. In the first experiment, the effects of different levels of N fertilizer (0 ; 20 ; 40 and 80 kg N/ha with or without liming were studied. Soybean podyield were related to N fertilization only when liming was added to the soil In the second one, the effects of four Bradyrhizobium japonicum strains F A3 ; 3-40 ; SA 1 and G3S on nodulation and yields were also studied. Inoculation has significant effect on nodulation and plant top dry weight at full bloom, and seed yield at harvest when compared to the control. However, the Bradyrhizobium japonicum strains tested had various symbiotic effectiveness on Glycine max cv. FN3. In addition, soybean plants inoculated with G3S strain and those fertilized with 100 kg N/ha produced similar seed yield. Our study illustrated that G3S strain had the better adaptability in environmental conditions of Congo soil.

  4. In vitro propagation of katsura tree (Cercidiphyllum japonicum Sieb ...

    African Journals Online (AJOL)

    ajl5

    compared the effects of phytohormone for seed germination and plant regeneration of katsura tree. Shoot .... produce new shoots (percentage of shoot initiation was. 0.66%), while all media supplemented with PGRs .... This probably indicated that bacterial in the buds are dormant. This is in agreement with the previous ...

  5. Biodiversity and biogeography of rhizobia associated with soybean plants grown in the North China Plain.

    Science.gov (United States)

    Zhang, Yan Ming; Li, Ying; Chen, Wen Feng; Wang, En Tao; Tian, Chang Fu; Li, Qin Qin; Zhang, Yun Zeng; Sui, Xin Hua; Chen, Wen Xin

    2011-09-01

    As the putative center of origin for soybean and the second largest region of soybean production in China, the North China Plain covers temperate and subtropical regions with diverse soil characteristics. However, the soybean rhizobia in this plain have not been sufficiently studied. To investigate the biodiversity and biogeography of soybean rhizobia in this plain, a total of 309 isolates of symbiotic bacteria from the soybean nodules collected from 16 sampling sites were studied by molecular characterization. These isolates were classified into 10 genospecies belonging to the genera Sinorhizobium and Bradyrhizobium, including four novel groups, with S. fredii (68.28%) as the dominant group. The phylogeny of symbiotic genes nodC and nifH defined four lineages among the isolates associated with Sinorhizobium fredii, Bradyrhizobium elkanii, B. japonicum, and B. yuanmingense, demonstrating the different origins of symbiotic genes and their coevolution with the chromosome. The possible lateral transfer of symbiotic genes was detected in several cases. The association between soil factors (available N, P, and K and pH) and the distribution of genospecies suggest clear biogeographic patterns: Sinorhizobium spp. were superdominant in sampling sites with alkaline-saline soils, while Bradyrhizobium spp. were more abundant in neutral soils. This study clarified the biodiversity and biogeography of soybean rhizobia in the North China Plain.

  6. Effect of bacterial root symbiosis and urea as source of nitrogen on performance of soybean plants grown hydroponically for bioregenerative life support systems (BLSSs

    Directory of Open Access Journals (Sweden)

    Roberta eParadiso

    2015-10-01

    Full Text Available Soybean is traditionally grown in soil, where root symbiosis with Bradyrhizobium japonicum can supply nitrogen (N, by means of bacterial fixation of atmospheric N2.Nitrogen fertilizers inhibit N-fixing bacteria. However urea is profitably used in soybean cultivation in soil, where urease enzymes of telluric microbes catalyze the hydrolysis to ammonium, which has a lighter inhibitory effect compared to nitrate.Previous researches demonstrated that soybean can be grown hydroponically with recirculating complete nitrate-based nutrient solutions. In Space, urea derived from crew urine could be used as N source, with positive effects in resource procurement and waste recycling. However, whether the plants are able to use urea as the sole source of N and its effect on root symbiosis with B. japonicum is still unclear in hydroponics.We compared the effect of two N sources, nitrate and urea, on plant growth and physiology, and seed yield and quality of soybean grown in closed-loop Nutrient Film Technique (NFT in growth chamber, with or without inoculation with B. japonicum.Urea limited plant growth and seed yield compared to nitrate by determining nutrient deficiency, due to its low utilization efficiency in the early developmental stages, and reduced nutrients uptake (K, Ca and Mg throughout the whole growing cycle. Root inoculation with B. japonicum did not improve plant performance, regardless of the N source. Specifically, nodulation increased under fertigation with urea compared to nitrate, but this effect did not result in higher leaf N content and better biomass and seed production.Urea was not suitable as sole N source for soybean in closed-loop NFT. However the ability to use urea increased from young to adult plants, suggesting the possibility to apply it during reproductive phase or in combination with nitrate in earlier developmental stages. Root symbiosis did not contribute significantly to N nutrition and did not enhance the plant ability

  7. Effect of bacterial root symbiosis and urea as source of nitrogen on performance of soybean plants grown hydroponically for Bioregenerative Life Support Systems (BLSSs).

    Science.gov (United States)

    Paradiso, Roberta; Buonomo, Roberta; Dixon, Mike A; Barbieri, Giancarlo; De Pascale, Stefania

    2015-01-01

    Soybean is traditionally grown in soil, where root symbiosis with Bradyrhizobium japonicum can supply nitrogen (N), by means of bacterial fixation of atmospheric N2. Nitrogen fertilizers inhibit N-fixing bacteria. However, urea is profitably used in soybean cultivation in soil, where urease enzymes of telluric microbes catalyze the hydrolysis to ammonium, which has a lighter inhibitory effect compared to nitrate. Previous researches demonstrated that soybean can be grown hydroponically with recirculating complete nitrate-based nutrient solutions. In Space, urea derived from crew urine could be used as N source, with positive effects in resource procurement and waste recycling. However, whether the plants are able to use urea as the sole source of N and its effect on root symbiosis with B. japonicum is still unclear in hydroponics. We compared the effect of two N sources, nitrate and urea, on plant growth and physiology, and seed yield and quality of soybean grown in closed-loop Nutrient Film Technique (NFT) in growth chamber, with or without inoculation with B. japonicum. Urea limited plant growth and seed yield compared to nitrate by determining nutrient deficiency, due to its low utilization efficiency in the early developmental stages, and reduced nutrients uptake (K, Ca, and Mg) throughout the whole growing cycle. Root inoculation with B. japonicum did not improve plant performance, regardless of the N source. Specifically, nodulation increased under fertigation with urea compared to nitrate, but this effect did not result in higher leaf N content and better biomass and seed production. Urea was not suitable as sole N source for soybean in closed-loop NFT. However, the ability to use urea increased from young to adult plants, suggesting the possibility to apply it during reproductive phase or in combination with nitrate in earlier developmental stages. Root symbiosis did not contribute significantly to N nutrition and did not enhance the plant ability to use

  8. Dicty_cDB: Contig-U13487-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available o rerio solute carrier family ... 63 5e-09 AE016879_1614( AE016879 |pid:none) Bacillus anthracis str. Ames, ... s... 46 0.44 4 ( DQ927303 ) Tetrahymena malaccensis strain MP75 mitochondrion... 40 0.44 4 ( AC174278 ) Medicago tr...-10 CU468135_2286( CU468135 |pid:none) Erwinia tasmaniensis strain ET1... 68 1e-10 CP001147_952( CP001147 |pid:non...pertussis strain 1... 49 9e-05 AF065159_25( AF065159 |pid:none) Bradyrhizobium japonicum ...e R3-3066P1, WO... 48 0.31 1 ( AE017355 ) Bacillus thuringiensis serovar konkukian str. 97-... 48 0.31 1

  9. Rhizosphere effect of colonizer plant species on the development of soil microbial community during primary succession on postmining sites

    Energy Technology Data Exchange (ETDEWEB)

    Elhottova, D.; Kristufek, V.; Maly, S.; Frouz, J. [Academy of Sciences of the Czech Republic, Ceske Budejovice (Czech Republic). Inst. for Soil Biology

    2009-07-01

    The impact of pioneer plant species Tussilago farfara on structural, functional, and growth characterization of microbial community colonizing the spoil colliery substrate was studied in a laboratory microcosm experiment. Microcosms consisting of spoil substrate (0.7 dm{sup 3} of tertiary alkaline clay sediment from Sokolov brown-coal mine area) from a pioneer site (without vegetation, 5 years after heaping) were cultivated in a greenhouse with one plant of this species. Plant roots substantially increased microbial diversity and biomass after one season (7 months) of cultivation. Roots influenced the microbial community and had nearly twice the size, higher growth, and metabolic potential in comparison to the control. The development of microbial specialists improves the plant nutrient status. Bacterial nitrogen (N{sub 2}) fixators (Bradyrhizobium japonicum, Rhizobium radiobacter) and arbuscular mycorrhizal fungi were confirmed in the rhizosphere of Tussilago farfara.

  10. Biotransformation of β-keto nitriles to chiral (S)-β-amino acids using nitrilase and ω-transaminase.

    Science.gov (United States)

    Mathew, Sam; Nadarajan, Saravanan Prabhu; Sundaramoorthy, Uthayasuriya; Jeon, Hyunwoo; Chung, Taeowan; Yun, Hyungdon

    2017-04-01

    To enzymatically synthesize enantiomerically pure β-amino acids from β-keto nitriles using nitrilase and ω-transaminase. An enzyme cascade system was designed where in β-keto nitriles are initially hydrolyzed to β-keto acids using nitrilase from Bradyrhizobium japonicum and subsequently β-keto acids were converted to β-amino acids using ω-transaminases. Five different ω-transaminases were tested for this cascade reaction, To enhance the yields of β-amino acids, the concentrations of nitrilase and amino donor were optimized. Using this enzymatic reaction, enantiomerically pure (S)-β-amino acids (ee > 99%) were generated. As nitrilase is the bottleneck in this reaction, molecular docking analysis was carried out to depict the poor affinity of nitrilase towards β-keto acids. A novel enzymatic route to generate enantiomerically pure aromatic (S)-β-amino acids from β-keto nitriles is demonstrated for the first time.

  11. Emisiones de óxido nitroso en un cultivo de soja [Glycine max (L. Merrill]: efecto de la inoculación y de la fertilización nitrógenada Nitrous oxide emission during a soybean [Glycine max (L. Merril] culture: inoculation and nitrogen fertilization effects

    Directory of Open Access Journals (Sweden)

    Ignacio A Ciampitti

    2005-12-01

    Full Text Available El óxido nitroso absorbe radiación infrarroja contribuyendo al efecto invernadero; este gas es producido principalmente en el suelo, mediante los procesos de nitrificación y denitrificación. En un estudio a campo, sobre un suelo Argiudol típico, se evaluó el efecto de la fertilización y la inoculación con Bradyrhizobium japonicum en un cultivo de soja [Glycine max (L. Merrill], sobre las emisiones de óxido nitroso. Los gases se extrajeron de cilindros de PVC y la lectura se realizó con cromatografía gaseosa. Las emisiones presentaron valores crecientes desde la siembra hacia madurez fisiológica del cultivo, para todos los tratamientos; este comportamiento fue concomitante con la evolución presentada por la humedad edáfica. La fertilización nitrogenada aumentó significativamente (PNitrous oxide gas absorbs infrared radiation contributing to the greenhouse effect; this gas is produced mainly in soil, by means of the processes of nitrification and denitrification. In a field study at the FAUBA on a typic Argiudol, we evaluated the effect of fertilization and inoculation with Bradyrhizobium japonicum during a soybean culture [Glycine max (L. Merrill], on nitrous oxide emisión. Gases were sampled with PVC cylinders and were read with gaseous chromatography. Emissions presented increasing values from seeding towards physiological maturity for all treatments; this behavior was similar to the evolution presented by nitrate level and soil moisture. Nitrogen fertilization significantly increased (P<0.05 nitrous oxide emissions and inoculation only had a significant effect with the highest level of fertilization (P=0.09. Plots fertilized at highest dose and inoculated gave the greatest nitrous oxide emissions. The variable that better explains the emissions is the nitrate level (r² = 0.1899; P=0.0231.

  12. Nodulação e fixação do dinitrogênio em soja tratada com sulfentrazone Nodulation and dinitrogen fixation in soybean treated with sulfentrazone

    Directory of Open Access Journals (Sweden)

    João Silveira Arruda

    2001-02-01

    Full Text Available Em condições de casa de vegetação, avaliou-se o efeito de doses (0, 36, 72, 108 e 144 µg m-2 de i.a. do herbicida sulfentrazone na nodulação da soja (Glycine max (L. Merrill cv. BR16 infectada com Bradyrhizobium japonicum estirpe SEMIA 5079, e na fixação de dinitrogênio, estimada pelos teores de ureídeos no exsudato do xilema. Foram quantificados, também, os teores de aminoácidos e nitrato no exsudato do xilema. O número e a matéria seca dos nódulos, as concentrações de aminoácidos, nitrato e ureídeos decresceram com o incremento na dose de sulfentrazone, tanto no estádio R3 quanto no R5 de desenvolvimento da soja.Under greenhouse conditions, the effect of sulfentrazone rates (0, 36, 72, 108 and 144 µg m-2 of a.i. was evaluated on growth of nodules in soybean (Glycine max (L. Merrill cv. BR16, inoculated with Bradyrhizobium japonicum strain SEMIA 5079, on dinitrogen fixation estimated by the content of ureides in xylem exsudate. The amino acids, nitrate and ureides contents in xylem exsudate were also determined. The number and dry matter of nodules, the content of amino acids, nitrate and ureides decreased as a function of increase of sulfentrazone rate at R3 and R5 development stages of soybean stages.

  13. Relationship between nitrogen sources, germination and vigor of soybean seeds Relações entre fontes de nitrogênio, germinação e vigor de sementes de soja

    Directory of Open Access Journals (Sweden)

    J. Marcos-Filho

    1994-08-01

    Full Text Available This research was conducted in two experimental areas; the first had been under cultivation with Bradyrhizobium japonicum anually inoculated for several years and the second was cultivated with sugarcane for many years receiving only mineral fertilization. Mineral nitrogen (urea and ammonium effect was compared with the effects of inoculant treatments (liquid and peat carriers on plant development, yield and seed quality. After harvesting, seeds were stored under normal environmental conditions and tested periodically. Germination, accelerated aging, electrical conductivity and seedling emergence tests showed that seeds coming from the area where inoculation was practiced have better quality. Best plant performance regarding plant height at different growth stages and yield was also found in this area. Among the nitrogen sources studied, best results were obtained when peat was the inoculant carrier.A pesquisa foi conduzida em duas áreas experimentais do Departamento de Agricultura, ESALQ/USP. A primeira, cultivada com soja anualmente inoculada com Bradyrhizobium japonicum, durante vários anos e, a segunda, anteriormente cultivada com cana-de-açúcar, recebendo apenas adubação mineral. Os efeitos da aplicação de uréia e de sulfato de amonio foram comparados com os da inoculação de sementes, utilizando-se, produtos veiculados em turfa ou em meio líquido; avaliaram-se o desenvolvimento das plantas, a produção final e a qualidade das sementes (testes de germinação e de vigor logo após a colheita e durante o armazenamento. Os resultados permitiram constatar o melhor desempenho fisiológico das sementes produzidas em área anteriormente cultivada com soja, o mesmo ocorrendo com o desenvolvimento inicial das plantas. Dentre as fontes de nitrogênio avaliadas, a utilização de inoculante veiculado em turfa se mostrou mais favorável.

  14. Bradyrhizobium populations occur in deep soil under the leguminous tree Acacia albida

    OpenAIRE

    Dupuy, Nicolas C.; Dreyfus, Bernard L.

    1992-01-01

    Soil cores were drilled under the leguminous tree Acacia albida growing in two different ecoclimatic zones of West Africa: the Sahelian area (100 to 500 mm of annual rainfall) and the Sudano-Guinean area (1,000 to 1,500 mm of annual rainfall). Soil samples were collected at different depths from the surface down to the water table level and analyzed for the presence of rhizobia able to nodulate A. albida. In both areas, population densities of rhizobia were substantially greater near the wate...

  15. Phenotypic and genotypic diversity of rhizobia nodulating Pterocarpus erinaceus and P. lucens in Senegal.

    Science.gov (United States)

    Sylla, Samba Ndao; Samba, Ramatoulaye Thiaba; Neyra, Marc; Ndoye, Ibrahima; Giraud, Eric; Willems, Anne; de Lajudie, Philippe; Dreyfus, Bernard

    2002-12-01

    A total of fifty root nodules isolates of fast-growing and slow growing rhizobia from Pterocarpus ennaceus and Pterocarpus lucens respectively native of sudanean and sahelian regions of Senegal were characterized. These isolates were compared to representative strains of known rhizobial species. Twenty-two new isolates were slow growers and twenty-eight were fast growers. A polyphasic approach was performed including comparative total protein sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE) profile analysis; 16S rDNA and 16S-23S rDNA intergenic spacer (IGS) sequence analysis. By SDS-PAGE the slow growing isolates grouped in one major cluster containing reference strains of Bradyrhizobium sp. including strains isolated in Africa, in Brazil and in New Zealand. Most of the fast-growing rhizobia grouped in four different clusters or were separate strains related to Rhizobium and Mesorhizobium strains. The 16S rDNA and 16S-23S rDNA IGS sequences analysis showed accurately the differentiation of fast growing rhizobia among the Rhizobium and Mesorbizobium genospecies. The representative strains of slow growing rhizobia were identified as closely related to Bradyrbizobium elkanii and Bradyrhizobium japonicum. Based on 16S rDNA sequence analysis, one slow growing strain (ORS199) was phylogenetically related to Bradyrbizobium sp. (Lupinus) and Blastobacter denitrificans. This position of ORS 199 was not confirmed by IGS sequence divergence. We found no clear relation between the diversity of strains, the host plants and the ecogeographical origins.

  16. Bradyrhizobia nodulating the Acacia mangium x A. auriculiformis interspecific hybrid are specific and differ from those associated with both parental species.

    Science.gov (United States)

    Le Roux, Christine; Tentchev, Diana; Prin, Yves; Goh, Doreen; Japarudin, Yani; Perrineau, Marie-Mathilde; Duponnois, Robin; Domergue, Odile; de Lajudie, Philippe; Galiana, Antoine

    2009-12-01

    In the context of an increasing utilization of the interspecific hybrid Acacia mangium x A. auriculiformis as a plantation tree in the tropical humid zone, its symbiotic characterization was carried out in comparison with that of its two parental species. Rhizobium strains of diverse geographical origins were isolated from root nodules of the hybrid and its parents. Almost all Acacia hybrid isolates were fast growing on yeast extract-mannitol medium, in contrast to those isolated from both parental species, which were mostly slow growing. The rhizobium strains were characterized through partial sequencing of the rRNA operon. In the phylogenetic tree, almost all strains isolated from the hybrid were grouped together in a clade close to Bradyrhizobium japonicum, while all strains isolated from both parental species were close to Bradyrhizobium elkanii. Inoculation experiments performed under in vitro or greenhouse conditions showed that all strains were infective with their original hosts but exhibited very variable degrees of effectivity according to the host plant tested. Thus, homologous strain-host associations were more effective than heterologous ones. This shows that there is still a high potential for isolating and testing new strains from hybrids to be used as inoculants in the context of large-scale afforestation programs.

  17. Bradyrhizobia Nodulating the Acacia mangium × A. auriculiformis Interspecific Hybrid Are Specific and Differ from Those Associated with Both Parental Species ▿ †

    Science.gov (United States)

    Le Roux, Christine; Tentchev, Diana; Prin, Yves; Goh, Doreen; Japarudin, Yani; Perrineau, Marie-Mathilde; Duponnois, Robin; Domergue, Odile; de Lajudie, Philippe; Galiana, Antoine

    2009-01-01

    In the context of an increasing utilization of the interspecific hybrid Acacia mangium × A. auriculiformis as a plantation tree in the tropical humid zone, its symbiotic characterization was carried out in comparison with that of its two parental species. Rhizobium strains of diverse geographical origins were isolated from root nodules of the hybrid and its parents. Almost all Acacia hybrid isolates were fast growing on yeast extract-mannitol medium, in contrast to those isolated from both parental species, which were mostly slow growing. The rhizobium strains were characterized through partial sequencing of the rRNA operon. In the phylogenetic tree, almost all strains isolated from the hybrid were grouped together in a clade close to Bradyrhizobium japonicum, while all strains isolated from both parental species were close to Bradyrhizobium elkanii. Inoculation experiments performed under in vitro or greenhouse conditions showed that all strains were infective with their original hosts but exhibited very variable degrees of effectivity according to the host plant tested. Thus, homologous strain-host associations were more effective than heterologous ones. This shows that there is still a high potential for isolating and testing new strains from hybrids to be used as inoculants in the context of large-scale afforestation programs. PMID:19854923

  18. Fe-phyllosilicate redox cycling organisms from a redox transition zone in Hanford 300 Area sediments

    Directory of Open Access Journals (Sweden)

    Jason eBenzine

    2013-12-01

    Full Text Available Microorganisms capable of reducing or oxidizing structural iron (Fe in Fe-bearing phyllosilicate minerals were enriched and isolated from a subsurface redox transition zone at the Hanford 300 Area site in eastern Washington, USA. Both conventional and in situ i-chip enrichment strategies were employed. One Fe(III-reducing Geobacter (G. bremensis strain R1, Deltaproteobacteria and six Fe(II phyllosilicate-oxidizing isolates from the Alphaproteobacteria (Bradyrhizobium japonicum strains 22, is5, and in8p8, Betaproteobacteria (Cupriavidus necator strain A5-1, Dechloromonas agitata strain is5, and Actinobacteria (Nocardioides sp. strain in31 were recovered. The G. bremensis isolate grew by oxidizing acetate with the oxidized form of NAu-2 smectite as the electron acceptor. The Fe(II-oxidizers grew by oxidation of chemically reduced smectite as the energy source with nitrate as the electron acceptor. The Bradyrhizobium isolates could also carry out aerobic oxidation of biotite. This is the first report of the recovery of a Fe(II-oxidizing Nocardioides, and to date only one other Fe(II-oxidizing Bradyrhizobium is known. The 16S rRNA gene sequences of the isolates were similar to ones found in clone libraries from Hanford 300 sediments and groundwater, suggesting that such organisms may be present and active in situ. Whole genome sequencing of the isolates is underway, the results of which will enable comparative genomic analysis of mechanisms of extracellular phyllosilicate Fe redox metabolism, and facilitate development of techniques to detect the presence and expression of genes associated with microbial phyllosilicate Fe redox cycling in sediments.

  19. The Soybean Rfg1 Gene Restricts Nodulation by Sinorhizobium fredii USDA193

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    Yinglun Fan

    2017-09-01

    Full Text Available Sinorhizobium fredii is a fast-growing rhizobial species that can establish a nitrogen-fixing symbiosis with a wide range of legume species including soybeans (Glycine max. In soybeans, this interaction shows a high level of specificity such that particular S. fredii strains nodulate only a limited set of plant genotypes. Here we report the identification of a dominant gene in soybeans that restricts nodulation with S. fredii USDA193. Genetic mapping in an F2 population revealed co-segregation of the underlying locus with the previously cloned Rfg1 gene. The Rfg1 allele encodes a member of the Toll-interleukin receptor/nucleotide-binding site/leucine-rich repeat class of plant resistance proteins that restricts nodulation by S. fredii strains USDA257 and USDA205, and an allelic variant of this gene also restricts nodulation by Bradyrhizobium japonicum USDA122. By means of complementation tests and CRISPR/Cas9-mediated gene knockouts, we demonstrate that the Rfg1 allele also is responsible for resistance to nodulation by S. fredii USDA193. Therefore, the Rfg1 allele likely provides broad-spectrum resistance to nodulation by many S. fredii and B. japonicum strains in soybeans.

  20. Distinct changes in soybean xylem sap proteome in response to pathogenic and symbiotic microbe interactions

    Directory of Open Access Journals (Sweden)

    Cho Un-Haing

    2009-09-01

    Full Text Available Abstract Background Plant systemic signaling characterized by the long distance transport of molecules across plant organs involves the xylem and phloem conduits. Root-microbe interactions generate systemic signals that are transported to aerial organs via the xylem sap. We analyzed the xylem sap proteome of soybean seedlings in response to pathogenic and symbiotic interactions to identify systemic signaling proteins and other differentially expressed proteins. Results We observed the increase of a serine protease and peroxidase in the xylem sap in response to Phytophthora sojae elicitor treatment. The high molecular weight fraction of soybean xylem sap was found to promote the growth of Neurospora crassa in vitro at lower concentrations and inhibit growth at higher concentrations. Sap from soybean plants treated with a P. sojae elicitor had a significantly higher inhibitory effect than sap from control soybean plants. When soybean seedlings were inoculated with the symbiont Bradyrhizobium japonicum, the abundance of a xyloglucan transendoglycosyl transferase protein increased in the xylem sap. However, RNAi-mediated silencing of the corresponding gene did not significantly affect nodulation in soybean hairy root composite plants. Conclusion Our study identified a number of sap proteins from soybean that are differentially induced in response to B. japonicum and P. sojae elicitor treatments and a majority of them were secreted proteins.

  1. Rizobacterias que promueven el desarrollo e incremento en productividad de Glycine max L.

    Directory of Open Access Journals (Sweden)

    Damián Antonio Cedeño Saavedra

    2017-06-01

    Full Text Available Los suelos agrícolas destinados al cultivo de especies vegetales de ciclo corto, carecen de materia orgánica por efecto del excesivo empleo de agroquímicos, estos fertilizantes edáficos de origen químico ocasionan cambios en la estructura y la biodiversidad del suelo. La incorporación de rizobacterias en la agricultura permite mejorar la productividad de cultivos de leguminosas por la formación de un complejo de simbiosis rhizobio-planta que permite la fijación de nitrógeno. El objetivo del presente trabajo fue evaluar el efecto de Bradyrhizobium japonicum, Pseudomonas veronii R4 y Pseudomonas fluorescens CHA0 para promover el desarrollo e incremento de productividad en tres variedades de soja: ICA-P34; INIAP-308; INIAP-307. A los 7 y 30 días se evaluó el desarrollo radicular y formación de nódulos, por co-inoculación de rizobacterias y sin inoculantes, también se determinó el rendimiento en kgha-1. En condiciones in vitro se incrementó el tejido radicular con 5, 3 y 9 cm de largo, por B. japonicum en ICA-P34, INIAP-308 y INIAP-307. En plántulas se observó raíces de 21 y 20 cm en INIAP-307 con P. veronii R4 y P. fluorescens CHA0. Inoculando las tres variedades con B. japonicum, se obtuvo 10, 14 y 10 nódulos por planta, cuando se aplicó Pseudomonas spp no existió formación de nódulos. El rendimiento de granos por hectárea fue 3700, 2890 y 3929 kgha-1, con B. japonicum, sin inoculo bacteriano los rendimientos disminuyen a 2006, 1611 y 1842 kgha-1. B. japonicum coloniza el sistema radicular influyendo con cambios morfológicos y fisiológicos, que positivamente incrementa el rendimiento en semillas de soja.

  2. Origin of a novel protein-coding gene family with similar signal sequence in Schistosoma japonicum

    National Research Council Canada - National Science Library

    Mbanefo, Evaristus Chibunna; Chuanxin, Yu; Kikuchi, Mihoko; Shuaibu, Mohammed Nasir; Boamah, Daniel; Kirinoki, Masashi; Hayashi, Naoko; Chigusa, Yuichi; Osada, Yoshio; Hamano, Shinjiro; Hirayama, Kenji

    2012-01-01

    ...). To find the mechanism underlying the origination of these genes with similar core promoter regions and signal sequence, we adopted an integrated approach utilizing whole genome, transcriptome...

  3. Efeitos do glifosato sobre microrganismos simbiotróficos de soja, em meio de cultura e casa de vegetação Effects of glyphosate on soybean symbiotic microorganisms, in culture media and in greenhouse

    Directory of Open Access Journals (Sweden)

    Juliano dos Santos Malty

    2006-02-01

    Full Text Available Os efeitos do herbicida Roundup, formulado à base de glifosato, foram avaliados sobre três estirpes de Bradyrhizobium elkanii (BR 29, INPA 80A e INPA 553A, e uma de B. japonicum (BR 86, e sobre três espécies de fungos micorrízicos arbusculares (FMA (Gigaspora margarita, Glomus etunicatum e Scutellospora heterogama, em meios de cultivo com concentrações crescentes do herbicida (0 a 454 µmol L-1; foram também avaliados os efeitos sobre a nodulação e micorrização da soja, em casa de vegetação, em solo que recebeu, antes da semeadura, doses do herbicida equivalentes a 1,25 até 10 L ha-1. O Roundup mostrou-se inibitório ao crescimento de Bradyrhizobium spp. e aos fungos em meio de cultura, e esse efeito foi crescente com o aumento das concentrações aplicadas, tendo variado em razão das espécies ou estirpes avaliadas. No entanto, a inibição in vitro só ocorreu em concentrações muito superiores à dose recomendada para aplicações no campo. As estirpes BR 29, INPA 553A e INPA 80A mostraram-se mais tolerantes ao glifosato, em relação à estirpe BR 86. O efeito do herbicida sobre a germinação e o crescimento dos tubos germinativos dos esporos dos FMA foi diferenciado, tendo sido observada inibição decrescente de G. etunicatum para S. heterogama e G. margarita. A aplicação do herbicida ao solo, antes da semeadura, até a dose equivalente a 10 L ha-1 não influenciou na nodulação e na colonização micorrízica da soja.The effects of the Roundup herbicide on three strains of Bradyrhizobium elkanii (BR 29, INPA 80A and INPA 553A, one of B. japonicum (BR 86, and on three species of arbuscular mycorrhizal fungi (AMF (Gigaspora margarita,Glomus etunicatum and Scutellospora heterogama, were evaluated in culture media containing increasing concentrations of the herbicide (0_454 µM; evaluations were also made on the effects on nodulation and mycorrhiza colonization of soybean grown in a soil treated with Roundup doses

  4. Conservation of Endangered Lupinus mariae-josephae in Its Natural Habitat by Inoculation with Selected, Native Bradyrhizobium Strains

    Science.gov (United States)

    Navarro, Albert; Fos, Simón; Laguna, Emilio; Durán, David; Rey, Luis; Rubio-Sanz, Laura; Imperial, Juan; Ruiz-Argüeso, Tomás

    2014-01-01

    Lupinus mariae-josephae is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain). In these soils, L. mariae-josephae grows in just a few defined patches, and previous conservation efforts directed towards controlled plant reproduction have been unsuccessful. We have previously shown that L. mariae-josephae plants establish a specific root nodule symbiosis with bradyrhizobia present in those soils, and we reasoned that the paucity of these bacteria in soils might contribute to the lack of success in reproducing plants for conservation purposes. Greenhouse experiments using L. mariae-josephae trap-plants showed the absence or near absence of L. mariae-josephae-nodulating bacteria in “terra rossa” soils of Valencia outside of L. mariae-josephae plant patches, and in other “terra rossa” or alkaline red soils of the Iberian Peninsula and Balearic Islands outside of the Valencia L. mariae-josephae endemism region. Among the bradyrhizobia able to establish an efficient symbiosis with L. mariae-josephae plants, two strains, LmjC and LmjM3 were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural L. mariae-josephae populations, and successful reproduction of the plant was achieved. Interestingly, the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia, and optimal performance was observed in plants inoculated with LmjC, a strain that had previously shown the most efficient behavior under controlled conditions. Our results define conditions for L. mariae-josephae conservation and for extension to alkaline-limed soil habitats, where no other known lupine can thrive. PMID:25019379

  5. Conservation of endangered Lupinus mariae-josephae in its natural habitat by inoculation with selected, native Bradyrhizobium strains.

    Science.gov (United States)

    Navarro, Albert; Fos, Simón; Laguna, Emilio; Durán, David; Rey, Luis; Rubio-Sanz, Laura; Imperial, Juan; Ruiz-Argüeso, Tomás

    2014-01-01

    Lupinus mariae-josephae is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain). In these soils, L. mariae-josephae grows in just a few defined patches, and previous conservation efforts directed towards controlled plant reproduction have been unsuccessful. We have previously shown that L. mariae-josephae plants establish a specific root nodule symbiosis with bradyrhizobia present in those soils, and we reasoned that the paucity of these bacteria in soils might contribute to the lack of success in reproducing plants for conservation purposes. Greenhouse experiments using L. mariae-josephae trap-plants showed the absence or near absence of L. mariae-josephae-nodulating bacteria in "terra rossa" soils of Valencia outside of L. mariae-josephae plant patches, and in other "terra rossa" or alkaline red soils of the Iberian Peninsula and Balearic Islands outside of the Valencia L. mariae-josephae endemism region. Among the bradyrhizobia able to establish an efficient symbiosis with L. mariae-josephae plants, two strains, LmjC and LmjM3 were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural L. mariae-josephae populations, and successful reproduction of the plant was achieved. Interestingly, the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia, and optimal performance was observed in plants inoculated with LmjC, a strain that had previously shown the most efficient behavior under controlled conditions. Our results define conditions for L. mariae-josephae conservation and for extension to alkaline-limed soil habitats, where no other known lupine can thrive.

  6. Conservation of endangered Lupinus mariae-josephae in its natural habitat by inoculation with selected, native Bradyrhizobium strains.

    Directory of Open Access Journals (Sweden)

    Albert Navarro

    Full Text Available Lupinus mariae-josephae is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain. In these soils, L. mariae-josephae grows in just a few defined patches, and previous conservation efforts directed towards controlled plant reproduction have been unsuccessful. We have previously shown that L. mariae-josephae plants establish a specific root nodule symbiosis with bradyrhizobia present in those soils, and we reasoned that the paucity of these bacteria in soils might contribute to the lack of success in reproducing plants for conservation purposes. Greenhouse experiments using L. mariae-josephae trap-plants showed the absence or near absence of L. mariae-josephae-nodulating bacteria in "terra rossa" soils of Valencia outside of L. mariae-josephae plant patches, and in other "terra rossa" or alkaline red soils of the Iberian Peninsula and Balearic Islands outside of the Valencia L. mariae-josephae endemism region. Among the bradyrhizobia able to establish an efficient symbiosis with L. mariae-josephae plants, two strains, LmjC and LmjM3 were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural L. mariae-josephae populations, and successful reproduction of the plant was achieved. Interestingly, the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia, and optimal performance was observed in plants inoculated with LmjC, a strain that had previously shown the most efficient behavior under controlled conditions. Our results define conditions for L. mariae-josephae conservation and for extension to alkaline-limed soil habitats, where no other known lupine can thrive.

  7. Conservation of Endangered Lupinus mariae-josephae in Its Natural Habitat by Inoculation with Selected, Native Bradyrhizobium Strains: e102205

    National Research Council Canada - National Science Library

    Albert Navarro; Simón Fos; Emilio Laguna; David Durán; Luis Rey; Laura Rubio-Sanz; Juan Imperial; Tomás Ruiz-Argüeso

    2014-01-01

      Lupinus mariae-josephae is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain). In these soils, L...

  8. Inoculating Acacia koa with Bradyrhizobium and applying fertilizer in the nursery: effects on nodule formation and seedling growth

    Science.gov (United States)

    R. Kasten Dumroese; Douglass F. Jacobs; Anthony S. Davis

    2009-01-01

    Restoration of Acacia koa A. Gray (koa) forests on degraded sites in Hawaii is important for conservation of rare, endemic plants and animals and is often accomplished by planting nursery-grown seedlings. To be successful after outplanting, koa seedlings must access sufficient nutrients from the soil and outcompete other vegetation. Forming symbiotic...

  9. Evaluation of the reaction oof interspecific hybrids of common bean and tepary bean to Bradyrhizobium y Rhizobium

    Science.gov (United States)

    Interspecific hybrids between common bean, Phaseolus vulgaris L., and tepary bean, Phaseolus acutifolius A. Gray, have the potential to increase bean production in regions where rainfall is limited. In 2014, an experiment was initiated using a split-plot design. The treatments included inoculation, ...

  10. Genetic diversity of indigenous rhizobial symbionts of the Lupinus mariae-josephae endemism from alkaline-limed soils within its area of distribution in Eastern Spain.

    Science.gov (United States)

    Durán, David; Rey, L; Sánchez-Cañizares, C; Navarro, A; Imperial, J; Ruiz-Argueso, T

    2013-03-01

    The genomic diversity of a collection of 103 indigenous rhizobia isolates from Lupinus mariae-josephae (Lmj), a recently described Lupinus species endemic to alkaline-limed soils from a restricted habitat in Eastern Spain, was investigated by molecular methods. Isolates were obtained from soils of four geographic locations in the Valencia province that harbored the known Lmj plant populations. Using an M13 RAPD fingerprinting technique, 19 distinct RAPD profiles were identified. Phylogenetic analysis based on 16S rDNA and the housekeeping genes glnII, recA and atpD showed a high diversity of native Bradyrhizobium strains that were able to establish symbiosis with Lmj. All the strains grouped in a clade unrelated to strains of the B. canariense and B. japonicum lineages that establish symbioses with lupines in acid soils of the Mediterranean area. The phylogenetic tree based on concatenated glnII, recA and atpD gene sequences grouped the Lmj isolates in six different operational taxonomic units (OTUs) at the 93% similarity level. These OTUs were not associated to any specific geographical location, and their observed divergence predicted the existence of different Bradyrhizobium genomic species. In contrast, phylogenetic analysis of symbiotic genes based on nodC and nodA gene sequences, defined only two distinct clusters among the Lmj strains. These two Lmj nod gene types were largely distinct from nod genes of bradyrhizobia nodulating other Old World lupine species. The singularity and large diversity of these strains in such a small geographical area makes this an attractive system for studying the evolution and adaptation of the rhizobial symbiont to the plant host. Copyright © 2012 Elsevier GmbH. All rights reserved.

  11. Transient Nod factor-dependent gene expression in the nodulation-competent zone of soybean (Glycine max [L.] Merr.) roots.

    Science.gov (United States)

    Hayashi, Satomi; Reid, Dugald E; Lorenc, Michał T; Stiller, Jiri; Edwards, David; Gresshoff, Peter M; Ferguson, Brett J

    2012-10-01

    All lateral organ development in plants, such as nodulation in legumes, requires the temporal and spatial regulation of genes and gene networks. A total mRNA profiling approach using RNA-seq to target the specific soybean (Glycine max) root tissues responding to compatible rhizobia [i.e. the Zone Of Nodulation (ZON)] revealed a large number of novel, often transient, mRNA changes occurring during the early stages of nodulation. Focusing on the ZON enabled us to discard the majority of root tissues and their developmentally diverse gene transcripts, thereby highlighting the lowly and transiently expressed nodulation-specific genes. It also enabled us to concentrate on a precise moment in early nodule development at each sampling time. We focused on discovering genes regulated specifically by the Bradyrhizobium-produced Nod factor signal, by inoculating roots with either a competent wild-type or incompetent mutant (nodC(-) ) strain of Bradyrhizobium japonicum. Collectively, 2915 genes were identified as being differentially expressed, including many known soybean nodulation genes. A number of unknown nodulation gene candidates and soybean orthologues of nodulation genes previously reported in other legume species were also identified. The differential expression of several candidates was confirmed and further characterized via inoculation time-course studies and qRT-PCR. The expression of many genes, including an endo-1,4-β-glucanase, a cytochrome P450 and a TIR-LRR-NBS receptor kinase, was transient, peaking quickly during the initiation of nodule ontogeny. Additional genes were found to be down-regulated. Significantly, a set of differentially regulated genes acting in the gibberellic acid (GA) biosynthesis pathway was discovered, suggesting a novel role of GAs in nodulation. © 2012 The Authors. Plant Biotechnology Journal © 2012 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

  12. EFEITOS DA INOCULAÇÃO COM BACTÉRIAS DIAZOTRÓFICAS E DA ADUBAÇÃO NITROGENADA NO CRESCIMENTO E NA QUALIDADE DE MUDAS DE Inga laurina (SW. Willd. (Fabaceae1

    Directory of Open Access Journals (Sweden)

    Gabriel Salles Góes

    2015-12-01

    Full Text Available Inga laurina é uma espécie arbórea com ampla distribuição na América do Sul, útil para sistemas agroflorestais, restauração florestal e arborização urbana. Este estudo teve como objetivo avaliar os efeitos da inoculação com bactérias fixadoras de nitrogênio (N e da adubação nitrogenada no crescimento e qualidade de mudas de I. laurina. O experimento teve duração de 170 dias, e as mudas foram cultivadas em tubetes plásticos com 115 cm3 de capacidade, contendo uma mistura 9: 1 em volume de HS Florestal® e pó de fibra de coco como substrato. Foram analisados seis tratamentos, sendo quatro inoculações (Bradyrhizobium japonicum 1 - BJ1, Rhizobium miluonense - RM, Bradyrhizobium japonicum 2- BJ2 e Burkholderia cepacia - BC, o controle positivo - C+ (sem inoculação e com adubação nitrogenada semanal, 60 mg dm3 de N, na forma de ureia e o controle negativo - C- (sem inoculação e sem adubação nitrogenada. Os tratamentos com inoculação foram pouco efetivos em relação ao crescimento das mudas, visto que as médias das variáveis de crescimento, da massa foliar específica (MFE e do índice de qualidade de Dixon (IQD foram significativamente superiores no C+, em comparação com os demais tratamentos. No entanto, os isolados RM e BJ2 foram efetivos na produção de nódulos, pois apresentaram os maiores valores médios da massa de matéria seca de nódulos (MSN. Além disso, os valores médios do índice de clorofila Falker (ICF foram significativamente superiores nos tratamentos com inoculação em relação ao C-. Os melhores resultados entre os tratamentos com inoculação foram obtidos para RM, seguido de BJ2.

  13. TRATAMENTO DE SEMENTES DE ARROZ, TRIGO, FEIJÃO E SOJA COM UM PRODUTO FORMULADO À BASE DE CÉLULAS E DE METABÓLITOS DE Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    E LAZZARETTI

    1997-01-01

    Full Text Available Um produto constituido de pó-molhável (PBBS formulado à base de células (60g e de metabólitos (60g de Bacillus subtilis, argila (480g, espalhante (7,92g e água (2400ml, moído e seco, mostrou-se semelhante aos tratamentos com fungicidas recomendados para o controle de Rhizoctonia solani, Aspergillus sp. e Sclerotinia sclerotiorum em sementes de feijão; Pyricularia oryzae e Rhinchosporium sativum em sementes de arroz; e Cercospora kikuchii, Phomopsis phaseoli e Fusarium spp. em sementes de soja. Para Dreschlera oryzae em arroz e Bipolaris sorokiniana, P. oryzae e Alternaria tenuis em sementes de trigo, o tratamento com o PBBS, embora não tenha se igualado ao tratamento com o fungicida padrão, diferiu estatisticamente do tratamento testemunha. A nodulação das raízes de feijão e soja por bactérias simbióticas, fixadoras de nitrogênio, não foi influenciada quando o PBBS foi aplicado simultaneamente ao inoculante contendo Rhyzobium sp. e Bradyrhizobium japonicum, respectivamente. O PBBS também não afetou a emergência das plântulas das culturas testadas.A biological fungicide product containing B. subtilis cells (60 g and metabolites (60g was transformed into a wettable powder formulated with, clay (480 g, surfactant (7,92 g and water (2400 ml.The product showed to be similar to the conventional fungicides used in the control of Rhizoctonia solani, Aspergillus sp. and Sclerotinia sclerotiorum in bean seeds. The control of Pyricularia oryzae and Rhinchosporium sativum in rice seeds and Cercospora kikuchii, Phomopsis phaseoli and Fusarium spp. in soybean seeds was also as efficient as the comercial fungicides. For Dreschlera oryzae in rice seeds and Bipolaris sorokiniana, P. oryzae and Alternaria tenuis in wheat seeds, the treatment with the product, althought not as efficient as the chemical treatment, was statistically distinct from the control. The root nodulation of bean and soybean by nitrogen fixing bacteria, was not affected

  14. Effectiveness of beneficial plant-microbe interactions under hypobaric and hypoxic conditions in an advanced life support system

    Science.gov (United States)

    MacIntyre, Olathe; Stasiak, Michael; Cottenie, Karl; Trevors, Jack; Dixon, Mike

    An assembled microbial community in the hydroponics solution of an advanced life support system may improve plant performance and productivity in three ways: (1) exclusion of plant pathogens from the initial community, (2) resistance to infection, and (3) plant-growth promotion. However, the plant production area is likely to have a hypobaric (low pressure) and hypoxic (low oxygen) atmosphere to reduce structural mass and atmosphere leakage, and these conditions may alter plant-microbe interactions. Plant performance and productivity of radish (Raphanus sativus L. cv. Cherry Bomb II) grown under hypobaric and hypoxic conditions were investigated at the University of Guelph's Controlled Environment Systems Research Facility. Changes in the microbial communities that routinely colonized the re-circulated nutrient solution, roots, and leaves of radishes in these experiments were quantified in terms of similarity in community composition, abundance of bacteria, and community diversity before and after exposure to hypobaric and hypoxic conditions relative to communities maintained at ambient growth conditions. The microbial succession was affected by extreme hypoxia (2 kPa oxygen partial pressure) while hypobaria as low as 10 kPa total pressure had little effect on microbial ecology. There were no correlations found between the physiological profile of these unintentional microbial communities and radish growth. The effects of hypobaric and hypoxic conditions on specific plant-microbe interactions need to be determined before beneficial gnotobiotic communities can be developed for use in space. The bacterial strains Tal 629 of Bradyrhizobium japonicum and WCS417 of Pseudomonas fluorescens, and the plant pathogen Fusarium oxysporum f. sp. raphani will be used in future experiments. B. japonicum Tal 629 promotes radish growth in hydroponics systems and P. fluorescens WCS417 induces systemic resistance to fusarium wilt (F. oxysporum f. sp. raphani) in radish under ambient

  15. Biophysical investigation of type A PutAs reveals a conserved core oligomeric structure

    Energy Technology Data Exchange (ETDEWEB)

    Korasick, David A. [Department of Biochemistry, University of Missouri, Columbia MO USA; Singh, Harkewal [Department of Chemistry, University of Missouri, Columbia MO USA; Pemberton, Travis A. [Department of Chemistry, University of Missouri, Columbia MO USA; Luo, Min [Department of Chemistry, University of Missouri, Columbia MO USA; Dhatwalia, Richa [Department of Chemistry, University of Missouri, Columbia MO USA; Tanner, John J. [Department of Biochemistry, University of Missouri, Columbia MO USA; Department of Chemistry, University of Missouri, Columbia MO USA

    2017-08-01

    Many enzymes form homooligomers, yet the functional significance of self-association is seldom obvious. Herein, we examine the connection between oligomerization and catalytic function for proline utilization A (PutA) enzymes. PutAs are bifunctional enzymes that catalyze both reactions of proline catabolism. Type A PutAs are the smallest members of the family, possessing a minimal domain architecture consisting of N-terminal proline dehydrogenase and C-terminal l-glutamate-γ-semialdehyde dehydrogenase modules. Type A PutAs form domain-swapped dimers, and in one case (Bradyrhizobium japonicum PutA), two of the dimers assemble into a ring-shaped tetramer. Whereas the dimer has a clear role in substrate channeling, the functional significance of the tetramer is unknown. To address this question, we performed structural studies of four-type A PutAs from two clades of the PutA tree. The crystal structure of Bdellovibrio bacteriovorus PutA covalently inactivated by N-propargylglycine revealed a fold and substrate-channeling tunnel similar to other PutAs. Small-angle X-ray scattering (SAXS) and analytical ultracentrifugation indicated that Bdellovibrio PutA is dimeric in solution, in contrast to the prediction from crystal packing of a stable tetrameric assembly. SAXS studies of two other type A PutAs from separate clades also suggested that the dimer predominates in solution. To assess whether the tetramer of B. japonicum PutA is necessary for catalytic function, a hot spot disruption mutant that cleanly produces dimeric protein was generated. The dimeric variant exhibited kinetic parameters similar to the wild-type enzyme. These results implicate the domain-swapped dimer as the core structural and functional unit of type A PutAs.

  16. Characterization of the flavonoid-responsive regulator FrrA and its binding sites.

    Science.gov (United States)

    Wenzel, Mandy; Lang, Kathrin; Günther, Tobias; Bhandari, Anita; Weiss, Andy; Lulchev, Pavel; Szentgyörgyi, Erik; Kranzusch, Bianca; Göttfert, Michael

    2012-05-01

    Previous microarray analyses revealed that in Bradyrhizobium japonicum, about 100 genes are induced by genistein, an isoflavonoid secreted by soybean. This includes the three genes freC, freA, and freB (systematic designations bll4319, bll4320, and bll4321), which are likely to form a genistein-, daidzein-, and coumestrol-inducible operon and to encode a multidrug efflux system. Upstream of freCAB and in the opposite orientation, FrrA (systematic designation Blr4322), which has similarity to TetR-type regulators, is encoded. A deletion of frrA leads to increased expression of freB in the absence of an inducer. We identified the correct translational start codon of frrA and showed that the gene is inducible by genistein and daidzein. The protein, which was heterologously expressed and purified from Escherichia coli, binds to two palindrome-like DNA elements (operator A and operator B), which are located in the intergenic region between freC and frrA. The replacement of several nucleotides or the insertion of additional spacer nucleotides prevented binding. Binding of FrrA was also affected by the addition of genistein. By mapping the transcription start sites, we found that operator A covers the transcriptional start site of freC and operator B is probably located between the -35 regions of the two divergently oriented genes. Operator A seems to be conserved in a few similar gene constellations in other proteobacteria. Our data indicate that in B. japonicum, besides NodD1 (the LysR family) and NodVW (a two-component response regulator), a third regulator type (a TetR family member) which responds to the plant signal molecules genistein and daidzein exists.

  17. Relationship between soil type and N₂O reductase genotype (nosZ) of indigenous soybean bradyrhizobia: nosZ-minus populations are dominant in Andosols.

    Science.gov (United States)

    Shiina, Yoko; Itakura, Manabu; Choi, Hyunseok; Saeki, Yuichi; Hayatsu, Masahito; Minamisawa, Kiwamu

    2014-01-01

    Bradyrhizobium japonicum strains that have the nosZ gene, which encodes N2O reductase, are able to mitigate N2O emissions from soils (15). To examine the distribution of nosZ genotypes among Japanese indigenous soybean bradyrhizobia, we isolated bradyrhizobia from the root nodules of soybean plants inoculated with 32 different soils and analyzed their nosZ and nodC genotypes. The 1556 resultant isolates were classified into the nosZ+/nodC+ genotype (855 isolates) and nosZ-/nodC+ genotype (701 isolates). The 11 soil samples in which nosZ- isolates significantly dominated (P volcanic ash soil prevalent in agricultural fields in Japan), whereas the 17 soil samples in which nosZ+ isolates significantly dominated were mainly alluvial soils (non-volcanic ash soils). This result was supported by a principal component analysis of environmental factors: the dominance of the nosZ- genotype was positively correlated with total N, total C, and the phosphate absorption coefficient in the soils, which are soil properties typical of Andosols. Internal transcribed spacer sequencing of representative isolates showed that the nosZ+ and nosZ- isolates of B. japonicum fell mainly into the USDA110 (BJ1) and USDA6 (BJ2) groups, respectively. These results demonstrated that the group lacking nosZ was dominant in Andosols, which can be a target soil type for an N2O mitigation strategy in soybean fields. We herein discussed how the nosZ genotypes of soybean bradyrhizobia depended on soil types in terms of N2O respiration selection and genomic determinants for soil adaptation.

  18. Relationship Between Soil Type and N2O Reductase Genotype (nosZ) of Indigenous Soybean Bradyrhizobia: nosZ-minus Populations are Dominant in Andosols

    Science.gov (United States)

    Shiina, Yoko; Itakura, Manabu; Choi, Hyunseok; Saeki, Yuichi; Hayatsu, Masahito; Minamisawa, Kiwamu

    2014-01-01

    Bradyrhizobium japonicum strains that have the nosZ gene, which encodes N2O reductase, are able to mitigate N2O emissions from soils (15). To examine the distribution of nosZ genotypes among Japanese indigenous soybean bradyrhizobia, we isolated bradyrhizobia from the root nodules of soybean plants inoculated with 32 different soils and analyzed their nosZ and nodC genotypes. The 1556 resultant isolates were classified into the nosZ+/nodC+ genotype (855 isolates) and nosZ−/nodC+ genotype (701 isolates). The 11 soil samples in which nosZ− isolates significantly dominated (P volcanic ash soil prevalent in agricultural fields in Japan), whereas the 17 soil samples in which nosZ+ isolates significantly dominated were mainly alluvial soils (non-volcanic ash soils). This result was supported by a principal component analysis of environmental factors: the dominance of the nosZ− genotype was positively correlated with total N, total C, and the phosphate absorption coefficient in the soils, which are soil properties typical of Andosols. Internal transcribed spacer sequencing of representative isolates showed that the nosZ+ and nosZ− isolates of B. japonicum fell mainly into the USDA110 (BJ1) and USDA6 (BJ2) groups, respectively. These results demonstrated that the group lacking nosZ was dominant in Andosols, which can be a target soil type for an N2O mitigation strategy in soybean fields. We herein discussed how the nosZ genotypes of soybean bradyrhizobia depended on soil types in terms of N2O respiration selection and genomic determinants for soil adaptation. PMID:25476067

  19. Solubilisation of inorganic phosphates by inoculant strains from tropical legumes

    Directory of Open Access Journals (Sweden)

    Leandro Marciano Marra

    2011-10-01

    Full Text Available Microbial solubilisation of low soluble inorganic phosphates is an important process contributing for the phosphorus available to plants in tropical soils. This study evaluates the ability of inoculant strains for tropical legumes to solubilise inorganic phosphates of low solubility that are found in tropical soils. Seven strains of Leguminosae nodulating bacteria (LNB were compared with one another and with a non-nodulating positive control, Burkholderia cepacia (LMG 1222T. Four of the strains are used as inoculants for cowpeas (Vigna unguiculata (Bradyrhizobium sp. UFLA 03-84; Bradyrhizobium elkani INPA 03-11B and Bradyrhizobium japonicum BR3267 or for common beans (Phaseolus vulgaris (Rhizobium tropici CIAT 899T. Rhizobium etli UFLA 02-100 and Rhizobium leguminosarum 316C10a are also efficient nodulators of beans and Cupriavidus taiwanensis LMG 19424T nodulates on Mimosa pudica. Two experiments, with solid and liquid media, were performed to determine whether the strains were able to solubilise CaHPO4, Al(H2PO43 or FePO4.2H2O. On solid GELP medium none of the strains dissolved FePO4.2H2O, but LMG 1222, UFLA 03-84 and CIAT 899 solubilised CaHPO4 particularly well. These strains, along with LMG 19424 and BR 3267, were also able to increase the solubility of Al(H2PO43. In liquid GELP medium, LMG 1222 solubilised all phosphate sources, but no legume nodulating strain could increase the solubility of Al(H2PO43. The strains CIAT 899 and UFLA 02-100 were the only legume nodulating bacteria able to solubilise the other phosphate sources in liquid media, dissolving both CaHPO4 and FePO4.2H2O. There was a negative correlation between the pH of the culture medium and the concentration of soluble phosphate when the phosphorus source was CaHPO4 or FePO4.2H2O. The contribution of these strains to increasing the phosphorus nutrition of legumes and non-legume plant species should be investigated further by in vivo experiments.

  20. Environ: E00579 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00579 Processed aconitum japonicum tuberous root Crude drug ... Aconitum japonicum, ...Aconitum [TAX:49188] ... Ranunculaceae (buttercup family) Aconitum japonicum tuberous root soaked in brine and coat with caustic lime (dried) ...

  1. Dicty_cDB: VFD143 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available |pid:none) Schistosoma japonicum phosphoglyce... 108 7e-23 FN315990_1( FN315990 |pid:none) Schistosoma japonicum...|pid:none) TSA: Schistosoma japonicum SJCHGC0... 108 7e-23 AY223444_1( AY223444 |pid:none) Schistosoma japonicum...|pid:none) Schistosoma japonicum isolate Anhu... 108 7e-23 FN315993_1( FN315993 |pid:none) Schistosoma japonicum

  2. Dicty_cDB: SLD883 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available |pid:none) Schistosoma japonicum phosphoglyce... 78 1e-13 FN315990_1( FN315990 |pid:none) Schistosoma japonicum...|pid:none) TSA: Schistosoma japonicum SJCHGC0... 78 1e-13 AY223444_1( AY223444 |pid:none) Schistosoma japonicum...|pid:none) Schistosoma japonicum isolate Anhu... 78 1e-13 FN315993_1( FN315993 |pid:none) Schistosoma japonicum

  3. Characterization of indigenous rhizobia from Caatinga

    Directory of Open Access Journals (Sweden)

    Fernanda Cíntia Pires e Teixeira

    2010-03-01

    Full Text Available The aim of this study was to characterize rhizobial isolates from Cratylia mollis Mart. ex Benth, Calliandra depauperata Benth. and Mimosa tenuiflora (Willd. Poir. by means of rhizobial colonies morphology and restriction analysis of the 16S ribosomal gene (16S rDNA-ARDRA. Nodules were collected in the field and from plants cultivated in a greenhouse experiment using Caatinga soil samples. Sixty seven isolates were described by morphological analysis. Forty seven representative isolates were used for ARDRA analysis using seven restriction enzymes. We observed high diversity of both slow and fast-growing rhizobia that formed three morpho-physiological clusters. A few fast-growing isolates formed a group of strains of the Bradyrhizobium type; however, most of them diverged from the B. japonicum and B. elkanii species. Cratylia mollis nodule isolates were the most diverse, while all Mimosa tenuiflora isolates displayed fast growth with no pH change and were clustered into groups bearing 100% similarity, according to ARDRA results.

  4. In situ stimulation vs. bioaugmentation: Can microbial inoculation of plant roots enhance biodegradation of organic compounds?

    Energy Technology Data Exchange (ETDEWEB)

    Kingsley, M.T.; Metting, F.B. Jr.; Fredrickson, J.K. [Pacific Northwest Lab., Richland, WA (United States); Seidler, R.J. [Environmental Protection Agency, Corvallis, OR (United States). Environmental Research Lab.

    1993-06-01

    The use of plant roots and their associated rhizosphere bacteria for biocontainment and biorestoration offers several advantages for treating soil-dispersed contaminants and for application to large land areas. Plant roots function as effective delivery systems, since root growth transports bacteria vertically and laterally along the root in the soil column (see [ 1,2]). Movement of microbes along roots and downward in the soil column can be enhanced via irrigation [1-4]. For example, Ciafardini et al. [3] increased the nodulation and the final yield of soybeans during pod filling by including Bradyrhizobium japonicum in the irrigation water. Using rhizosphere microorganisms is advantageous for biodegradation of compounds that are degraded mainly by cometabolic processes, e.g., trichloroethylene (TCE). The energy source for bacterial growth and metabolism is supplied by the plant in the form of root exudates and other sloughed organic material. Plants are inexpensive, and by careful choice of species that possess either tap or fibrous root growth patterns, they can be used to influence mass transport of soil contaminants to the root surface via the transpiration stream [5]. Cropping of plants to remove heavy metals from contaminated soils has been proposed as a viable, low-cost, low-input treatment option [6]. The interest in use of plants as a remediation strategy has even reached the popular press [7], where the use of ragweed for the reclamation of sites contaminated with tetraethyl lead and other heavy metals was discussed.

  5. Tn5-Mob transposon mediated transfer of salt tolerance and symbiotic characteristics between Rhizobia genera.

    Science.gov (United States)

    Yang, S; Wu, Z; Gao, W; Li, J

    1993-01-01

    Rhizobium meliloti 042B is a fast-growing, salt-tolerant and high efficiency nitrogen-fixing symbiont with alfalfa. Bradyrhizobium japonicum USDA110 grows slowly, and cannot grow in YMA medium containing 0.1M NaCl, but nodulates and fixed nitrogen efficiently with soybean. Eighty-six transconjugants, called SR, were obtained by inserting Tn5-Mob randomly into genomes of 042B using pSUP5011 and helper plasmid RP4. Selecting 4 SR strains randomly and introducing DNA fragment of SR into USDA110 with helper plasmid R68.45 by triparental mating, 106 transconjugants, called BSR, were constructed. Most of BSR strains had the fast-growing phenotype and could tolerate 0.3-0.5M NaCl generally. Some of them produced melanine. When soybean and alfalfa were inoculated with these transconjugants BSR, 47 out of 90 BSR were found to nodulate in both of these plants, but no nitrogenase activity was observed with alfalfa; 26 strains could only nodulate and fix nitrogen in soybean; 13 strains could nodulate in alfalfa but did not fix nitrogen; 4 strains failed to nodulate in either soybean or alfalfa. Among them, 4 transconjugants which tolerated and fixed nitrogen efficiently in soybean were constructed.

  6. Growth Response of Two Phaseolus mungo L. Cultivars Induced by Arbuscular Mycorrhizal Fungi and Trichoderma viride

    Directory of Open Access Journals (Sweden)

    Navnita Sharma

    2016-01-01

    Full Text Available The present investigation aimed to quantify the difference in response of two Phaseolus mungo L. cultivars (i.e., UH-1 and IPU-94-1 to Glomus mosseae (G, that is, Funneliformis mosseae, Acaulospora laevis (A, and Trichoderma viride (T, in different combinations or alone. All the treatments were inoculated with Bradyrhizobium japonicum to ensure nodulation as soil used in the experiment was sterilized. After 120 days of inoculation, plants were analyzed for chlorophyll content, nodulation, mycorrhization, leaf area, and protein content. Results indicate variation in growth response of two cultivars with different treatments. Triple inoculation of plants with G + A + T proved to be the best treatment for growth followed by G + T in both cultivars. Our work allowed the selection of P. mungo L. cultivar UH-1 as highly mycorrhizal responsive as compared to IPU-94-1 and G. mosseae to be an efficient bioinoculant as compared to A. laevis for growth enhancement of P. mungo. Further characterization of P. mungo genotypes will enhance our knowledge of physiological and genetic mechanism behind increase in plant growth and yield due to AM symbiosis.

  7. Protein oxidation mediated by heme-induced active site conversion specific for heme-regulated transcription factor, iron response regulator.

    Science.gov (United States)

    Kitatsuji, Chihiro; Izumi, Kozue; Nambu, Shusuke; Kurogochi, Masaki; Uchida, Takeshi; Nishimura, Shin-ichiro; Iwai, Kazuhiro; O'Brian, Mark R; Ikeda-Saito, Masao; Ishimori, Koichiro

    2016-01-05

    The Bradyrhizobium japonicum transcriptional regulator Irr (iron response regulator) is a key regulator of the iron homeostasis, which is degraded in response to heme binding via a mechanism that involves oxidative modification of the protein. Here, we show that heme-bound Irr activates O2 to form highly reactive oxygen species (ROS) with the "active site conversion" from heme iron to non-heme iron to degrade itself. In the presence of heme and reductant, the ROS scavenging experiments show that Irr generates H2O2 from O2 as found for other hemoproteins, but H2O2 is less effective in oxidizing the peptide, and further activation of H2O2 is suggested. Interestingly, we find a time-dependent decrease of the intensity of the Soret band and appearance of the characteristic EPR signal at g = 4.3 during the oxidation, showing the heme degradation and the successive formation of a non-heme iron site. Together with the mutational studies, we here propose a novel "two-step self-oxidative modification" mechanism, during which O2 is activated to form H2O2 at the heme regulatory motif (HRM) site and the generated H2O2 is further converted into more reactive species such as ·OH at the non-heme iron site in the His-cluster region formed by the active site conversion.

  8. Magnesium-dependent processes are targets of bacterial manganese toxicity

    Science.gov (United States)

    Hohle, Thomas H.; O’Brian, Mark R.

    2014-01-01

    A Bradyrhizobium japonicum mutant defective in the gene encoding the high affinity Mn2+ transporter MntH has a severe growth phenotype under manganese limitation. Here, we isolated suppressor mutants of an mntH strain that grew under manganese limitation, and activities of high affinity Mn2+ transport and Mn2+-dependent enzymes were partially rescued. The suppressor strains harbor gain-of-function mutations in the gene encoding the Mg2+ channel MgtE. The MgtE variants likely allow Mn2+ entry via loss of a gating mechanism that normally holds the transporter in the closed state when cellular Mg2+ levels are high. Both MgtE-dependent and -independent suppressor phenotypes were recapitulated by magnesium-limited growth of the mntH strain. Growth studies of wild type cells suggest that manganese is toxic to cells when environmental magnesium is low. Moreover, extracellular manganese and magnesium levels were manipulated to inhibit growth without substantially altering the intracellular content of either metal, implying that manganese toxicity depends on its cellular distribution rather than the absolute concentration. Mg2+-dependent enzyme activities were found to be inhibited or stimulated by Mn2+. We conclude that Mn2+ can occupy Mg2+-binding sites in cells, and suggest that Mg2+-dependent processes are targets of manganese toxicity. PMID:24975873

  9. Structure and Biological Roles of Sinorhizobium fredii HH103 Exopolysaccharide

    Science.gov (United States)

    Acosta-Jurado, Sebastián; Soto, María J.; Margaret, Isabel; Crespo-Rivas, Juan C.; Sanjuan, Juan; Temprano, Francisco; Gil-Serrano, Antonio; Ruiz-Sainz, José E.; Vinardell, José M.

    2014-01-01

    Here we report that the structure of the Sinorhizobium fredii HH103 exopolysaccharide (EPS) is composed of glucose, galactose, glucuronic acid, pyruvic acid, in the ratios 5∶2∶2∶1 and is partially acetylated. A S. fredii HH103 exoA mutant (SVQ530), unable to produce EPS, not only forms nitrogen fixing nodules with soybean but also shows increased competitive capacity for nodule occupancy. Mutant SVQ530 is, however, less competitive to nodulate Vigna unguiculata. Biofilm formation was reduced in mutant SVQ530 but increased in an EPS overproducing mutant. Mutant SVQ530 was impaired in surface motility and showed higher osmosensitivity compared to its wild type strain in media containing 50 mM NaCl or 5% (w/v) sucrose. Neither S. fredii HH103 nor 41 other S. fredii strains were recognized by soybean lectin (SBL). S. fredii HH103 mutants affected in exopolysaccharides (EPS), lipopolysaccharides (LPS), cyclic glucans (CG) or capsular polysaccharides (KPS) were not significantly impaired in their soybean-root attachment capacity, suggesting that these surface polysaccharides might not be relevant in early attachment to soybean roots. These results also indicate that the molecular mechanisms involved in S. fredii attachment to soybean roots might be different to those operating in Bradyrhizobium japonicum. PMID:25521500

  10. (Catalytic mechanism of hydrogenase from aerobic N sub 2 -fixing microorganisms)

    Energy Technology Data Exchange (ETDEWEB)

    Arp, D.J.

    1990-01-01

    Hydrogenases are enzymes which catalyze reactions involving dihydrogen. They serve integral roles in a number of microbial metabolic pathways. Our research is focussed on investigations of the catalytic mechanism of the hydrogenases found in aerobic, N{sub 2}-fixing microorganisms such as Azotobacter vinelandii and the agronomically important Bradyrhizobium japonicum as well as microorganisms with similar hydrogenases. The hydrogenases isolated from these microorganisms are Ni- and Fe-containing heterodimers. Our work has focussed on three areas during the last grant period. In all cases, a central theme has been the role of inhibitors in the characteristics under investigation. In addition, a number of collaborative efforts have yielded interesting results. In metalloenzymes such as hydrogenase, inhibitors often influence the activity of the enzyme through ligand interactions with redox centers, often metals, within the enzyme. Therefore, investigations of the ability of various compounds to inhibit an enzyme's activity, as well as the mechanism of inhibition, can provide insight into the catalytic mechanism of the enzyme as well as the role of various redox centers in catalysis. We have investigated in detail four inhibitors of A. vinelandii and the results are summarized here. The influence of these inhibitors on the spectral properties of the enzyme are summarized. Electron paramagnetic resonance and ultraviolet spectra investigations are discussed. 9 figs.

  11. Structure and biological roles of Sinorhizobium fredii HH103 exopolysaccharide.

    Directory of Open Access Journals (Sweden)

    Dulce N Rodríguez-Navarro

    Full Text Available Here we report that the structure of the Sinorhizobium fredii HH103 exopolysaccharide (EPS is composed of glucose, galactose, glucuronic acid, pyruvic acid, in the ratios 5∶2∶2∶1 and is partially acetylated. A S. fredii HH103 exoA mutant (SVQ530, unable to produce EPS, not only forms nitrogen fixing nodules with soybean but also shows increased competitive capacity for nodule occupancy. Mutant SVQ530 is, however, less competitive to nodulate Vigna unguiculata. Biofilm formation was reduced in mutant SVQ530 but increased in an EPS overproducing mutant. Mutant SVQ530 was impaired in surface motility and showed higher osmosensitivity compared to its wild type strain in media containing 50 mM NaCl or 5% (w/v sucrose. Neither S. fredii HH103 nor 41 other S. fredii strains were recognized by soybean lectin (SBL. S. fredii HH103 mutants affected in exopolysaccharides (EPS, lipopolysaccharides (LPS, cyclic glucans (CG or capsular polysaccharides (KPS were not significantly impaired in their soybean-root attachment capacity, suggesting that these surface polysaccharides might not be relevant in early attachment to soybean roots. These results also indicate that the molecular mechanisms involved in S. fredii attachment to soybean roots might be different to those operating in Bradyrhizobium japonicum.

  12. Evolution of acyl-substrate recognition by a family of acyl-homoserine lactone synthases.

    Directory of Open Access Journals (Sweden)

    Quin H Christensen

    Full Text Available Members of the LuxI protein family catalyze synthesis of acyl-homoserine lactone (acyl-HSL quorum sensing signals from S-adenosyl-L-methionine and an acyl thioester. Some LuxI family members prefer acyl-CoA, and others prefer acyl-acyl carrier protein (ACP as the acyl-thioester substrate. We sought to understand the evolutionary history and mechanisms mediating this substrate preference. Our phylogenetic and motif analysis of the LuxI acyl-HSL synthase family indicates that the acyl-CoA-utilizing enzymes evolved from an acyl-ACP-utilizing ancestor. To further understand how acyl-ACPs and acyl-CoAs are recognized by acyl-HSL synthases we studied BmaI1, an octanoyl-ACP-dependent LuxI family member from Burkholderia mallei, and BjaI, an isovaleryl-CoA-dependent LuxI family member from Bradyrhizobium japonicum. We synthesized thioether analogs of their thioester acyl-substrates to probe recognition of the acyl-phosphopantetheine moiety common to both acyl-ACP and acyl-CoA substrates. The kinetics of catalysis and inhibition of these enzymes indicate that they recognize the acyl-phosphopantetheine moiety and they recognize non-preferred substrates with this moiety. We find that CoA substrate utilization arose through exaptation of acyl-phosphopantetheine recognition in this enzyme family.

  13. Preinoculation of Soybean Seeds Treated with Agrichemicals up to 30 Days before Sowing: Technological Innovation for Large-Scale Agriculture

    Science.gov (United States)

    da Cruz, Sonia Purin; Martin, Thomas Newton; Nakatani, André Shigueyoshi; Nogueira, Marco Antonio; Hungria, Mariangela

    2017-01-01

    The cultivation of soybean in Brazil experienced an expressive growth in the last decades. Soybean is highly demanding on nitrogen (N) that must come from fertilizers or from biological fixation. The N supply to the soybean crop in Brazil relies on the inoculation with elite strains of Bradyrhizobium japonicum, B. elkanii, and B. diazoefficiens, which are able to fulfill the crop's N requirements and enrich the soil for the following crop. The effectiveness of the association between N2-fixing bacteria and soybean plants depends on the efficacy of the inoculation process. Seed treatment with pesticides, especially fungicides or micronutrients, may rapidly kill the inoculated bacteria, affecting the establishment and outcome of the symbiosis. The development of technologies that allow inoculation to become a successful component of industrial seed treatment represents a valuable tool for the seed industry, as well as for the soybean crop worldwide. In this article, we report the results of new technologies, developed by the company Total Biotecnologia Indústria e Comércio S/A of Brazil, for preinoculation of soybean seeds with bradyrhizobia, in the presence of agrichemicals. Our results demonstrate improved bacterial survival for up to 30 days after inoculation, without compromising nodulation, N2-fixation, and yield in the field. PMID:29129977

  14. Elucidation of gibberellin biosynthesis in bacteria reveals convergent evolution.

    Science.gov (United States)

    Nett, Ryan S; Montanares, Mariana; Marcassa, Ariana; Lu, Xuan; Nagel, Raimund; Charles, Trevor C; Hedden, Peter; Rojas, Maria Cecilia; Peters, Reuben J

    2017-01-01

    Gibberellins (GAs) are crucial phytohormones involved in many aspects of plant growth and development, including plant-microbe interactions, which has led to GA production by plant-associated fungi and bacteria as well. While the GA biosynthetic pathways in plants and fungi have been elucidated and found to have arisen independently through convergent evolution, little has been uncovered about GA biosynthesis in bacteria. Some nitrogen-fixing, symbiotic, legume-associated rhizobia, including Bradyrhizobium japonicum-the symbiont of soybean-and Sinorhizobium fredii-a broad-host-nodulating species-contain a putative GA biosynthetic operon, or gene cluster. Through functional characterization of five unknown genes, we demonstrate that this operon encodes the enzymes necessary to produce GA9, thereby elucidating bacterial GA biosynthesis. The distinct nature of these enzymes indicates that bacteria have independently evolved a third biosynthetic pathway for GA production. Furthermore, our results also reveal a central biochemical logic that is followed in all three convergently evolved GA biosynthetic pathways.

  15. Preinoculation of Soybean Seeds Treated with Agrichemicals up to 30 Days before Sowing: Technological Innovation for Large-Scale Agriculture

    Directory of Open Access Journals (Sweden)

    Ricardo Silva Araujo

    2017-01-01

    Full Text Available The cultivation of soybean in Brazil experienced an expressive growth in the last decades. Soybean is highly demanding on nitrogen (N that must come from fertilizers or from biological fixation. The N supply to the soybean crop in Brazil relies on the inoculation with elite strains of Bradyrhizobium japonicum, B. elkanii, and B. diazoefficiens, which are able to fulfill the crop’s N requirements and enrich the soil for the following crop. The effectiveness of the association between N2-fixing bacteria and soybean plants depends on the efficacy of the inoculation process. Seed treatment with pesticides, especially fungicides or micronutrients, may rapidly kill the inoculated bacteria, affecting the establishment and outcome of the symbiosis. The development of technologies that allow inoculation to become a successful component of industrial seed treatment represents a valuable tool for the seed industry, as well as for the soybean crop worldwide. In this article, we report the results of new technologies, developed by the company Total Biotecnologia Indústria e Comércio S/A of Brazil, for preinoculation of soybean seeds with bradyrhizobia, in the presence of agrichemicals. Our results demonstrate improved bacterial survival for up to 30 days after inoculation, without compromising nodulation, N2-fixation, and yield in the field.

  16. X-ray microanalytical studies of mineral elements in the tripartite symbiosis between lima bean, N2-fixing bacteria and mycorrhizal fungi.

    Science.gov (United States)

    Rodak, Bruna Wurr; Freitas, Douglas Siqueira; Bamberg, Soraya Marx; Carneiro, Marco Aurélio Carbone; Guilherme, Luiz Roberto Guimarães

    2017-01-01

    The symbiosis between legumes, arbuscular mycorrhizal (AM) fungi, and N 2 -fixing bacteria (NFB) provides mutual nutritional gains. However, assessing the nutritional status of the microorganisms is a difficult task. A methodology that could assess this status, in situ, could assist managing these organisms in agriculture. This study used X-ray microanalyses to quantify and locate mineral elements in structures formed in a tripartite symbiosis. Lima bean (Phaseolus lunatus L. Walp) was cultivated in pots under greenhouse conditions, to which we have added AM fungal isolates (Glomus macrocarpum and Acaulospora colombiana) and NFB (Bradyrhizobium japonicum) inocula. Uninoculated control plants were also included. Symbionts were evaluated at the onset of flowering. Quantification of the mineral elements in the symbiotic components was performed using energy dispersive X-ray spectroscopy (EDX) and a scanning electron microscopy (SEM) was used to identify structures. EDX analysis detected 13 elements with the most abundant being N, Ca, and Se, occurring in all tissues, Fe in roots, Ni and Al in epidermis and P and Mo in nodules. Elemental quantification in fungal structures was not possible. The distribution of elements was related to their symbiotic function. X-ray microanalysis can be efficiently applied for nutritional diagnosis in tripartite symbiosis. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Preinoculation of Soybean Seeds Treated with Agrichemicals up to 30 Days before Sowing: Technological Innovation for Large-Scale Agriculture.

    Science.gov (United States)

    Araujo, Ricardo Silva; da Cruz, Sonia Purin; Souchie, Edson Luiz; Martin, Thomas Newton; Nakatani, André Shigueyoshi; Nogueira, Marco Antonio; Hungria, Mariangela

    2017-01-01

    The cultivation of soybean in Brazil experienced an expressive growth in the last decades. Soybean is highly demanding on nitrogen (N) that must come from fertilizers or from biological fixation. The N supply to the soybean crop in Brazil relies on the inoculation with elite strains of Bradyrhizobium japonicum, B. elkanii, and B. diazoefficiens, which are able to fulfill the crop's N requirements and enrich the soil for the following crop. The effectiveness of the association between N2-fixing bacteria and soybean plants depends on the efficacy of the inoculation process. Seed treatment with pesticides, especially fungicides or micronutrients, may rapidly kill the inoculated bacteria, affecting the establishment and outcome of the symbiosis. The development of technologies that allow inoculation to become a successful component of industrial seed treatment represents a valuable tool for the seed industry, as well as for the soybean crop worldwide. In this article, we report the results of new technologies, developed by the company Total Biotecnologia Indústria e Comércio S/A of Brazil, for preinoculation of soybean seeds with bradyrhizobia, in the presence of agrichemicals. Our results demonstrate improved bacterial survival for up to 30 days after inoculation, without compromising nodulation, N2-fixation, and yield in the field.

  18. Molluscicidal activities of medicinal plants from eastern China against Oncomelania hupensis, the intermediate host of Schistosoma japonicum Atividades moluscicida de plantas medicinais do leste da China contra Oncomelania hupensis, o hospedeiro intermediário da Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    Bang-xing Han

    2010-11-01

    Full Text Available In a search for natural products that could be used to control schistosomiasis, nineteen extracts of eleven medicinal plants from eastern China have been tested for molluscicidal activity against snail Oncomelania hupensis. The n-butanol fraction of the fresh leaf from Buddleja lindleyana Fortune, Buddlejaceae, showed significant activity against the snails. Statistical analysis revealed that the LC50 and LC90 values for the n-butanol fraction were 39.91 mg L-1 and 59.28 mg L-1 for 48 h, respectively. Otherwise, the LC50 values for the n-butanol fraction to zebrafish was 403.24 mg L-1 for 48 h. Therefore, the n-butanol fraction of the fresh leaf from B. lindleyana may be a potent and safe molluscicides.Na busca por produtos naturais que podem ser utilizados para controle da esquistossomose, dezenove extratos de onze plantas medicinais do leste da China foram testados para atividade moluscicida contra o caramujo Oncomelania hupensis. A fração n-butanol das folhas frescas de Buddleja lindleyana Fortune, Buddlejaceae, mostrou atividade significativa contra os caracóis. A análise estatística revelou que os valores de CL50 e CL90 para a fração n-butanol foram 39,91 mg L-1 e 59,28 mg L-1 por 48 h, respectivamente. Por outro lado, a CL50 para a fração n-butanol para peixe-zebra foi 403,24 mg L-1 por 48 h. Portanto, a fração n-butanol das folhas frescas de B. lindleyana poderá vir a ser um moluscicidas potente e seguro.

  19. Local immune responses of the Chinese water buffalo, Bubalus bubalis, against Schistosoma japonicum larvae: crucial insights for vaccine design.

    Directory of Open Access Journals (Sweden)

    Hamish E G McWilliam

    Full Text Available Asian schistosomiasis is a zoonotic parasitic disease infecting up to a million people and threatening tens of millions more. Control of this disease is hindered by the animal reservoirs of the parasite, in particular the water buffalo (Bubalus bubalis, which is responsible for significant levels of human transmission. A transmission-blocking vaccine administered to buffaloes is a realistic option which would aid in the control of schistosomiasis. This will however require a better understanding of the immunobiology of schistosomiasis in naturally exposed buffaloes, particularly the immune response to migrating schistosome larvae, which are the likely targets of an anti-schistosome vaccine. To address this need we investigated the immune response at the major sites of larval migration, the skin and the lungs, in previously exposed and re-challenged water buffaloes. In the skin, a strong allergic-type inflammatory response occurred, characterised by leukocyte and eosinophil infiltration including the formation of granulocytic abscesses. Additionally at the local skin site, interleukin-5 transcript levels were elevated, while interleukin-10 levels decreased. In the skin-draining lymph node (LN a predominant type-2 profile was seen in stimulated cells, while in contrast a type-1 profile was detected in the lung draining LN, and these responses occurred consecutively, reflecting the timing of parasite migration. The intense type-2 immune response at the site of cercarial penetration is significantly different to that seen in naive and permissive animal models such as mice, and suggests a possible mechanism for immunity. Preliminary data also suggest a reduced and delayed immune response occurred in buffaloes given high cercarial challenge doses compared with moderate infections, particularly in the skin. This study offers a deeper understanding into the immunobiology of schistosomiasis in a natural host, which may aid in the future design of more effective vaccines.

  20. Formation and Controlled Drug Release Using a Three-Component Supramolecular Hydrogel for Anti-Schistosoma Japonicum Cercariae

    Directory of Open Access Journals (Sweden)

    Yibao Li

    2016-03-01

    Full Text Available A novel three-component supramolecular hydrogel based on riboflavin, melamine and amino acid derivatives were constructed for controlled release of pesticides, Niclosamide derivatives. The formation of hydrogel may be attributed to self-assemble via hydrogen bonding and π–π interaction, which have been researched via scanning electron microscopy (SEM and Fourier transform infrared (FT-IR spectra. The rheological experiments showed that the hydrogel materials and drug-loaded hydrogel all demonstrated good mechanical strength and high stability. Further experimental results indicated that the drug-loaded hydrogels show large drug loadings, long-term release time and relatively higher efficiency to anti-cercariae in the water environment.

  1. Molluscicidal activities of medicinal plants from eastern China against Oncomelania hupensis, the intermediate host of Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    Bang-xing Han

    2010-09-01

    Full Text Available In a search for natural products that could be used to control schistosomiasis, nineteen extracts of eleven medicinal plants from eastern China have been tested for molluscicidal activity against snail Oncomelania hupensis. The n-butanol fraction of the fresh leaf from Buddleja lindleyana Fortune, Buddlejaceae, showed significant activity against the snails. Statistical analysis revealed that the LC50 and LC90 values for the n-butanol fraction were 39.91 mg L-1 and 59.28 mg L-1 for 48 h, respectively. Otherwise, the LC50 values for the n-butanol fraction to zebrafish was 403.24 mg L-1 for 48 h. Therefore, the n-butanol fraction of the fresh leaf from B. lindleyana may be a potent and safe molluscicides.

  2. Green synthesis of silver nanoparticles using Ganoderma neo-japonicum Imazeki: a potential cytotoxic agent against breast cancer cells

    OpenAIRE

    Gurunathan, Sangiliyandi; Raman, Jegadeesh; Malek, Sri Nurestri Abd; John, Priscilla A; Vikineswary, Sabaratnam

    2013-01-01

    Sangiliyandi Gurunathan,1 Jegadeesh Raman,2 Sri Nurestri Abd Malek,2 Priscilla A John,2 Sabaratnam Vikineswary2 1Department of Animal Biotechnology, Konkuk University, Seoul, South Korea; 2Mushroom Research Centre, University of Malaya, Kuala Lumpur, Malaysia Background: Silver nanoparticles (AgNPs) are an important class of nanomaterial for a wide range of industrial and biomedical applications. AgNPs have been used as antimicrobial and disinfectant agents due their detrimental effect on ta...

  3. NCBI nr-aa BLAST: CBRC-TTRU-01-1326 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1326 emb|CAX75571.1| Transmembrane protein 118 [Schistosoma japonicum]... emb|CAX75572.1| Transmembrane protein 118 [Schistosoma japonicum] emb|CAX75574.1| Transmembrane protein 118 [Schistosoma japon...icum] emb|CAX75575.1| Transmembrane protein 118 [Schistosoma japonicum] CAX75571.1 0.36 27% ...

  4. NCBI nr-aa BLAST: CBRC-MMUR-01-0199 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-0199 emb|CAX76110.1| putative tubulin, beta, 2 [Schistosoma japonicum]... emb|CAX76111.1| putative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76112.1| putative tubulin, beta, 2 [Schistosoma japon...icum] emb|CAX76113.1| putative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76114.1| p...utative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76115.1| putative tubulin..., beta, 2 [Schistosoma japonicum] emb|CAX76116.1| putative tubulin, beta, 2 [Schistosoma japonicum] CAX76110.1 2e-31 82% ...

  5. Dicty_cDB: Contig-U13988-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SJECDG02 SJE Schistosoma japonicum cDNA, mRNA seq... 36 0.16 2 ( BU769607 ) SJECGH07 SJE Schistosoma japonicum...SJEADF12 SJE Schistosoma japonicum cDNA, mRNA seq... 36 0.16 2 ( BU767943 ) SJEBEE09 SJE Schistosoma japonicum...SJEAPC08 SJE Schistosoma japonicum cDNA, mRNA seq... 36 0.16 2 ( BU772583 ) SJEETF03 SJE Schistosoma japonicum...SJECWA01 SJE Schistosoma japonicum cDNA, mRNA seq... 36 0.16 2 ( BU774006 ) SJEGBG02 SJE Schistosoma japonicum...DQ076155 ) Schistosoma japonicum clone 42E5 unknown mRNA. 36 0.16 2 ( BU768246 ) SJEBHG10 SJE Schistosoma japonicum

  6. NCBI nr-aa BLAST: CBRC-DNOV-01-0421 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0421 ref|NP_066199.2| ATP synthase F0 subunit 6 [Schistosoma japonicum...] gb|AAG13139.2| ATPase subunit 6 [Schistosoma japonicum] gb|AAL12132.1| ATPase subunit 6 [Schistosoma japon...icum] gb|AAL12140.1| ATPase subunit 6 [Schistosoma japonicum] gb|AAL12148.1| ATPase subunit 6 [Schistosoma japonicum] NP_066199.2 2.9 23% ...

  7. High-quality permanent draft genome sequence of Bradyrhizobium sp. Tv2a.2, a microsymbiont of Tachigali versicolor discovered in Barro Colorado Island of Panama.

    Science.gov (United States)

    Tian, Rui; Parker, Matthew; Seshadri, Rekha; Reddy, Tbk; Markowitz, Victor; Ivanova, Natalia; Pati, Amrita; Woyke, Tanja; Baeshen, Mohammed N; Baeshen, Nabih A; Kyrpides, Nikos; Reeve, Wayne

    2015-01-01

    Bradyrhizobiumsp. Tv2a.2 is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from an effective nitrogen-fixing root nodule of Tachigali versicolor collected in Barro Colorado Island of Panama. Here we describe the features of Bradyrhizobiumsp. Tv2a.2, together with high-quality permanent draft genome sequence information and annotation. The 8,496,279 bp high-quality draft genome is arranged in 87 scaffolds of 87 contigs, contains 8,109 protein-coding genes and 72 RNA-only encoding genes. This rhizobial genome was sequenced as part of the DOE Joint Genome Institute 2010 Genomic Encyclopedia for Bacteria and Archaea-Root Nodule Bacteria (GEBA-RNB) project.

  8. Classificação taxonômica das estirpes de rizóbio recomendadas para as culturas da soja e do feijoeiro baseada no seqüenciamento do gene 16S rRNA Taxonomic classification of rhizobial strains recommended for soybean and common bean crops in Brazil based on the sequencing of the 16s rRNA gene

    Directory of Open Access Journals (Sweden)

    L. M. O. Chueire

    2003-10-01

    Full Text Available As culturas da soja [Glycine max (L. Merrill] e do feijoeiro (Phaseolus vulgaris L. são de grande importância econômica e social para o Brasil e ambas podem ter seu requerimento de nitrogênio suprido pela simbiose com bactérias da ordem Rhizobiales. Para garantir a maximização do processo biológico, deve-se proceder à inoculação de estirpes de rizóbio eficientes e competitivas, recomendadas pela pesquisa. No Brasil, foram comercializados, na safra 2001/2002, 14 milhões de doses de inoculantes, dos quais 99 % para as culturas da soja e do feijoeiro. Neste trabalho, determinou-se a posição taxonômica das estirpes utilizadas em inoculantes comerciais para as duas culturas, pelo seqüenciamento da região do DNA que codifica o gene 16S rRNA, que é suficientemente variável, mas carrega as informações necessárias para permitir a análise filogenética de bactérias. O seqüenciamento permitiu definir que duas das estirpes recomendadas para a cultura da soja, SEMIA 587 e SEMIA 5019 (= 29 w, pertencem à espécie Bradyrhizobium elkanii e as duas outras, SEMIA 5079 (=CPAC 15 e SEMIA 5080 (= CPAC 7, à espécie B. japonicum. Determinou-se, ainda, que a estirpe SEMIA 4080 (=PRF 81, recomendada para o cultura do feijoeiro, pertence à espécie Rhizobium tropici. As seqüências obtidas foram depositadas no banco mundial de genes do National Center for Biotechnology Information.Soybean [Glycine max (L. Merrill] and common bean (Phaseolus vulgaris L. crops are of economical and social importance in Brazil; their requirement for nitrogen can be supplied by the symbiosis with bacteria belonging to the order Rhizobiales. However, to guarantee the maximization of the biological nitrogen fixation, seeds must be inoculated with efficient and competitive strains of rhizobia recommended by research. In 2001/2002, 14 million doses of inoculant were sold in Brazil, 99 % of these for soybean and common bean crops. In this study the taxonomic

  9. Influence of fungicide seed treatment on soybean nodulation and grain yield Influência do tratamento de sementes com fungicidas na nodulação e rendimento de grãos da soja

    Directory of Open Access Journals (Sweden)

    Jerri Édson Zilli

    2009-08-01

    Full Text Available Biological N2 fixation is a major factor contributing to the increased competitiveness of Brazilian soybeans on the international market. However, the contribution of this process may be limited by adverse conditions to symbiotic bacteria, such as fungicide seed treatments. This study aimed to evaluate the effects of the fungicides carbendazim + thiram and carboxin + thiram on soybean nodulation, plant growth and grain yield. Two field experiments were carried out in the Cerrado region of the State of Roraima, in a soil with a low organic matter content and no soybean bradyrhizobia. In 2005, seeds were treated with fungicide carbendazim + thiram and commercial inoculants containing the Bradyrhizobium elkanii strains SEMIA 5019 and SEMIA 587 and B. japonicum strains SEMIA 5079 and SEMIA 5080. In 2006, soybean seeds were treated with the fungicides carbendazim + thiram or carboxin + thiram and inoculated separately with each one of the four strains. The plants were evaluated for number of nodules and dry weight, shoot dry weight and total N accumulated in shoots 35 days after plant emergence, while grain yield and N grain content were determined at harvest. Both fungicides reduced soybean nodulation, especially in the presence of B. elkanii strains. The fungicide carbendazim + thiram reduced nodulation by about 50 % and grain yield by more than 20 % (about 700 kg ha-1, in the treatment inoculated with of strain SEMIA 587.A fixação biológica de N2 representa um dos principais fatores que aumentam a competitividade da soja no mercado internacional. Entretanto, a resposta desse processo pode ser limitada por condições adversas à bactéria, como o tratamento de sementes com fungicidas. O objetivo deste estudo foi avaliar o efeito de fungicidas à base de carbendazim + tiram e carboxin + tiram sobre a nodulação, o desenvolvimento das plantas e o rendimento de grãos da cultura da soja. Dois experimentos foram realizados no Cerrado do Estado de

  10. Adubação nitrogenada suplementar tardia em soja cultivada em latossolos do Cerrado Late supplemental nitrogen fertilization on soybean cropped in Cerrado Oxisols

    Directory of Open Access Journals (Sweden)

    Iêda de Carvalho Mendes

    2008-08-01

    Full Text Available O objetivo deste trabalho foi avaliar se a adubação nitrogenada, nos estádios de pré-florescimento (R1 e início do enchimento de grãos (R5, interfere na produtividade e na nodulação da soja. O estudo foi realizado por sete anos, em Latossolo Vermelho-Amarelo argiloso e, por quatro anos, em Latossolo Vermelho argiloso, ambos com populações estabelecidas de Bradyrhizobium. Foram conduzidos 15 ensaios, em plantio direto ou convencional, com os seguintes tratamentos: inoculação-padrão (IP com B. japonicum; IP + 200 kg ha-1 de N (uréia parcelados em 50% na semeadura e 50% em R1; IP + 50 kg ha-1 de N (nitrato de amônio em R1; IP + 50 kg ha-1 de N (sulfato de amônio em R1; IP + 50 kg ha-1 de N (nitrato de amônio em R5; e IP + 50 kg ha-1 de N (sulfato de amônio em R5. A aplicação de 200 kg ha-1 de N prejudicou a nodulação. Em apenas dois ensaios, houve resposta à aplicação de N, no entanto, sem retorno econômico ao produtor. A adubação nitrogenada tardia, no cultivo da soja com inoculação, em latossolos do Cerrado, não se justifica economicamente, em nenhum dos sistemas de cultivo avaliados, independentemente da fonte de N utilizada.The objective of this work was to evaluate the effects of nitrogen fertilization at the pre-flowering (R1 and seed filling (R5 stages, on the nodulation and yield of soybean grown in Cerrado Oxisols. The study was performed for seven years in a clayey Typic Haplustox, and for four years in a clayey Rhodic Haplustox, both with established populations of Bradyrhizobium. Fifteen experiments were carried out under conventional and no tillage management systems and the treatments were: standard inoculation (SI with B. japonicum; SI + 200 kg ha-1 N (urea, split-applied 50% at sowing and 50% at the R1 stage; SI + 50 kg ha-1 N (ammonium nitrate in R1; SI + 50 kg ha-1 N (ammonium sulfate in R1; SI + 50 kg ha-1 N (ammonium nitrate in R5; and SI + 50 kg ha-1 N (ammonium sulfate in R5. The application of

  11. Absence of Ancient DNA in Sub-Fossil Insect Inclusions Preserved in ‘Anthropocene’ Colombian Copal

    Science.gov (United States)

    Penney, David; Wadsworth, Caroline; Fox, Graeme; Kennedy, Sandra L.; Preziosi, Richard F.; Brown, Terence A.

    2013-01-01

    Insects preserved in copal, the sub-fossilized resin precursor of amber, have potential value in molecular ecological studies of recently-extinct species and of extant species that have never been collected as living specimens. The objective of the work reported in this paper was therefore to determine if ancient DNA is present in insects preserved in copal. We prepared DNA libraries from two stingless bees (Apidae: Meliponini: Trigonisca ameliae) preserved in ‘Anthropocene’ Colombian copal, dated to ‘post-Bomb’ and 10,612±62 cal yr BP, respectively, and obtained sequence reads using the GS Junior 454 System. Read numbers were low, but were significantly higher for DNA extracts prepared from crushed insects compared with extracts obtained by a non-destructive method. The younger specimen yielded sequence reads up to 535 nucleotides in length, but searches of these sequences against the nucleotide database revealed very few significant matches. None of these hits was to stingless bees though one read of 97 nucleotides aligned with two non-contiguous segments of the mitochondrial cytochrome oxidase subunit I gene of the East Asia bumblebee Bombus hypocrita. The most significant hit was for 452 nucleotides of a 470-nucleotide read that aligned with part of the genome of the root-nodulating bacterium Bradyrhizobium japonicum. The other significant hits were to proteobacteria and an actinomycete. Searches directed specifically at Apidae nucleotide sequences only gave short and insignificant alignments. All of the reads from the older specimen appeared to be artefacts. We were therefore unable to obtain any convincing evidence for the preservation of ancient DNA in either of the two copal inclusions that we studied, and conclude that DNA is not preserved in this type of material. Our results raise further doubts about claims of DNA extraction from fossil insects in amber, many millions of years older than copal. PMID:24039876

  12. Absence of ancient DNA in sub-fossil insect inclusions preserved in 'Anthropocene' Colombian copal.

    Science.gov (United States)

    Penney, David; Wadsworth, Caroline; Fox, Graeme; Kennedy, Sandra L; Preziosi, Richard F; Brown, Terence A

    2013-01-01

    Insects preserved in copal, the sub-fossilized resin precursor of amber, have potential value in molecular ecological studies of recently-extinct species and of extant species that have never been collected as living specimens. The objective of the work reported in this paper was therefore to determine if ancient DNA is present in insects preserved in copal. We prepared DNA libraries from two stingless bees (Apidae: Meliponini: Trigonisca ameliae) preserved in 'Anthropocene' Colombian copal, dated to 'post-Bomb' and 10,612±62 cal yr BP, respectively, and obtained sequence reads using the GS Junior 454 System. Read numbers were low, but were significantly higher for DNA extracts prepared from crushed insects compared with extracts obtained by a non-destructive method. The younger specimen yielded sequence reads up to 535 nucleotides in length, but searches of these sequences against the nucleotide database revealed very few significant matches. None of these hits was to stingless bees though one read of 97 nucleotides aligned with two non-contiguous segments of the mitochondrial cytochrome oxidase subunit I gene of the East Asia bumblebee Bombus hypocrita. The most significant hit was for 452 nucleotides of a 470-nucleotide read that aligned with part of the genome of the root-nodulating bacterium Bradyrhizobium japonicum. The other significant hits were to proteobacteria and an actinomycete. Searches directed specifically at Apidae nucleotide sequences only gave short and insignificant alignments. All of the reads from the older specimen appeared to be artefacts. We were therefore unable to obtain any convincing evidence for the preservation of ancient DNA in either of the two copal inclusions that we studied, and conclude that DNA is not preserved in this type of material. Our results raise further doubts about claims of DNA extraction from fossil insects in amber, many millions of years older than copal.

  13. Prediction and overview of the RpoN-regulon in closely related species of the Rhizobiales.

    Science.gov (United States)

    Dombrecht, Bruno; Marchal, Kathleen; Vanderleyden, Jos; Michiels, Jan

    2002-01-01

    In the rhizobia, a group of symbiotic Gram-negative soil bacteria, RpoN (sigma54, sigmaN, NtrA) is best known as the sigma factor enabling transcription of the nitrogen fixation genes. Recent reports, however, demonstrate the involvement of RpoN in other symbiotic functions, although no large-scale effort has yet been undertaken to unravel the RpoN-regulon in rhizobia. We screened two complete rhizobial genomes (Mesorhizobium loti, Sinorhizobium meliloti) and four symbiotic regions (Rhizobium etli, Rhizobium sp. NGR234, Bradyrhizobium japonicum, M. loti) for the presence of the highly conserved RpoN-binding sites. A comparison was also made with two closely related non-symbiotic members of the Rhizobiales (Agrobacterium tumefaciens, Brucella melitensis). A highly specific weight-matrix-based screening method was applied to predict members of the RpoN-regulon, which were stored in a highly annotated and manually curated dataset. Possible enhancer-binding proteins (EBPs) controlling the expression of RpoN-dependent genes were predicted with a profile hidden Markov model. The methodology used to predict RpoN-binding sites proved highly effective as nearly all known RpoN-controlled genes were identified. In addition, many new RpoN-dependent functions were found. The dependency of several of these diverse functions on RpoN seems species-specific. Around 30% of the identified genes are hypothetical. Rhizobia appear to have recruited RpoN for symbiotic processes, whereas the role of RpoN in A. tumefaciens and B. melitensis remains largely to be elucidated. All species screened possess at least one uncharacterized EBP as well as the usual ones. Lastly, RpoN could significantly broaden its working range by direct interfering with the binding of regulatory proteins to the promoter DNA.

  14. Molecular and microscopic analysis of the gut contents of abundant rove beetle species (Coleoptera, Staphylinidae in the boreal balsam fir forest of Quebec, Canada

    Directory of Open Access Journals (Sweden)

    Jan Klimaszewski

    2013-11-01

    Full Text Available Experimental research on beetle responses to removal of logging residues following clearcut harvesting in the boreal balsam fir forest of Quebec revealed several abundant rove beetle (Staphylinidae species potentially important for long-term monitoring. To understand the trophic affiliations of these species in forest ecosystems, it was necessary to analyze their gut contents. We used microscopic and molecular (DNA methods to identify the gut contents of the following rove beetles: Atheta capsularis Klimaszew­ski, Atheta klagesi Bernhauer, Oxypoda grandipennis (Casey, Bryophacis smetanai Campbell, Ischnosoma longicorne (Mäklin, Mycetoporus montanus Luze, Tachinus frigidus Erichson, Tachinus fumipennis (Say, Tachinus quebecensis Robert, and Pseudopsis subulata Herman. We found no apparent arthropod fragments within the guts; however, a number of fungi were identified by DNA sequences, including filamentous fungi and budding yeasts [Ascomycota: Candida derodonti Suh & Blackwell (accession number FJ623605, Candida mesenterica (Geiger Diddens & Lodder (accession number FM178362, Candida railenensis Ramirez and Gonzáles (accession number JX455763, Candida sophie-reginae Ramirez & González (accession number HQ652073, Candida sp. (accession number AY498864, Pichia delftensis Beech (accession number AY923246, Pichia membranifaciens Hansen (accession number JQ26345, Pichia misumaiensis Y. Sasaki and Tak. Yoshida ex Kurtzman 2000 (accession number U73581, Pichia sp. (accession number AM261630, Cladosporium sp. (accession number KF367501, Acremonium psammosporum W. Gams (accession number GU566287, Alternaria sp. (accession number GU584946, Aspergillus versicolor Bubak (accession number AJ937750, and Aspergillus amstelodami (L. Mangin Thom and Church (accession number HQ728257]. In addition, two species of bacteria [Bradyrhizobium japonicum (KirchnerJordan (accession number BA000040 and Serratia marcescens Bizio accession number CP003942] were found in

  15. Phytotoxicity attenuation in Vigna radiata under heavy metal stress at the presence of biochar and N fixing bacteria.

    Science.gov (United States)

    Seneviratne, Mihiri; Weerasundara, Lakshika; Ok, Yong Sik; Rinklebe, Jörg; Vithanage, Meththika

    2017-01-15

    This study assesses the effect of N-fixing bacteria and biochar synergism on plant growth and development of Vigna mungo under heavy metal stress (HM). Heavy metal stress is a worldwide problem, which causes critical effects on plant life due to oxidative stress. Application of biochar is a recent biological remediation technique, which often leads to an immobilization of heavy metals in soil. . Synergism of bacteria and biochar is a novel aspect to enhance plant growth under heavy metal stress. Woody biochar a byproduct of a dendro power industry was added as 1, 2.5 and 5% amounts combination with Bradyrhizobium japonicum, where mung seedlings were planted in serpentine soil rich in Ni, Mn, Cr and Co. Pot experiments were conducted for 12 weeks. The plant height, heavy metal uptake by plants, soil bioavailable heavy metal contents, soil N and P and microbial biomass carbon (MBC) were measured. The plant growth was enhanced with biochar amendment but a retardation was observed with high biochar application (5%). The soil N and P increased with the increase of biochar addition percentage while soil MBC showed reductions at 5% biochar amendment. Both soil bioavailable fractions of HM and up take of HMs by plants were gradually reduced with increase in biochar content. Based on the results, 2.5% biochar synergism with bacteria was the best for plant growth and soil nutrition status. Despite the synergism, available N was negatively correlated with the decrease of bioavailable metal percentage in soil whereas it was conversely for P. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Laser-ablation electrospray ionization mass spectrometry with ion mobility separation reveals metabolites in the symbiotic interactions of soybean roots and rhizobia

    Energy Technology Data Exchange (ETDEWEB)

    Stopka, Sylwia A.; Agtuca, Beverly J.; Koppenaal, David W.; Pasa Tolic, Ljiljana; Stacey, Gary; Vertes, Akos; Anderton, Christopher R.

    2017-05-23

    Technologies enabling in situ metabolic profiling of living plant systems are invaluable for understanding physiological processes and could be used for rapid phenotypic screening (e.g., to produce plants with superior biological nitrogen fixing ability). The symbiotic interaction between legumes and nitrogen-fixing soil bacteria results in a specialized plant organ (i.e., root nodule), where the exchange of nutrients between host and endosymbiont occurs. Laser ablation electrospray ionization mass spectrometry (LAESI-MS) is a method that can be performed under ambient conditions requiring minimal sample preparation. Here, we employed LAESI-MS to explore the well-characterized symbiosis between soybean (Glycine max L. Merr.) and its compatible symbiont, Bradyrhizobium japonicum. The utilization of ion mobility separation (IMS) improved the molecular coverage, selectivity, and identification of the detected biomolecules. Specifically, incorporation of IMS resulted in an increase of 153 detected metabolites in the nodule samples. The data presented demonstrates the advantages of using LAESI-IMS-MS for the rapid analysis of intact root nodules, uninfected root segments, and free-living rhizobia. Untargeted pathway analysis revealed several metabolic processes within the nodule (e.g., zeatin, riboflavin, and purine synthesis). Compounds specific to the uninfected root and bacteria were also detected. Lastly, we performed depth-profiling of intact nodules to reveal the location of metabolites to the cortex and inside the infected region, and lateral profiling of sectioned nodules confirmed these molecular distributions. Our results established the feasibility of LAESI-IMS-MS for the analysis and spatial mapping of plant tissues, with its specific demonstration to improve our understanding of the soybean-rhizobial symbiosis.

  17. Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria.

    Directory of Open Access Journals (Sweden)

    Seiya Watanabe

    Full Text Available Opines, in particular nopaline and octopine, are specific compounds found in crown gall tumor tissues induced by infections with Agrobacterium species, and are synthesized by well-studied NAD(PH-dependent dehydrogenases (synthases, which catalyze the reductive condensation of α-ketoglutarate or pyruvate with L-arginine. The corresponding genes are transferred into plant cells via a tumor-inducing (Ti plasmid. In addition to the reverse oxidative reaction(s, the genes noxB-noxA and ooxB-ooxA are considered to be involved in opine catabolism as (membrane-associated oxidases; however, their properties have not yet been elucidated in detail due to the difficulties associated with purification (and preservation. We herein successfully expressed Nox/Oox-like genes from Pseudomonas putida in P. putida cells. The purified protein consisted of different α-, β-, and γ-subunits encoded by the OdhA, OdhB, and OdhC genes, which were arranged in tandem on the chromosome (OdhB-C-A, and exhibited dehydrogenase (but not oxidase activity toward nopaline in the presence of artificial electron acceptors such as 2,6-dichloroindophenol. The enzyme contained FAD, FMN, and [2Fe-2S]-iron sulfur as prosthetic groups. On the other hand, the gene cluster from Bradyrhizobium japonicum consisted of OdhB1-C-A-B2, from which two proteins, OdhAB1C and OdhAB2C, appeared through the assembly of each β-subunit together with common α- and γ-subunits. A poor phylogenetic relationship was detected between OdhB1 and OdhB2 in spite of them both functioning as octopine dehydrogenases, which provided clear evidence for the acquisition of novel functions by "subunit-exchange". To the best of our knowledge, this is the first study to have examined flavin-containing opine dehydrogenase.

  18. Genome sequence of the chemolithoautotrophic nitrite-oxidizing bacterium Nitrobacter winogradskyi Nb-255

    Energy Technology Data Exchange (ETDEWEB)

    Hauser, Loren John [ORNL; Land, Miriam L [ORNL; Larimer, Frank W [ORNL; Arp, D J [Oregon State University; Hickey, W J [University of Wisconsin, Madison

    2006-03-01

    The alphaproteobacterium Nitrobacter winogradskyi (ATCC 25391) is a gram-negative facultative chemolithoautotroph capable of extracting energy from the oxidation of nitrite to nitrate. Sequencing and analysis of its genome revealed a single circular chromosome of 3,402,093 bp encoding 3,143 predicted proteins. There were extensive similarities to genes in two alphaproteobacteria, Bradyrhizobium japonicum USDA110 (1,300 genes) and Rhodopseudomonas palustris CGA009 CG (815 genes). Genes encoding pathways for known modes of chemolithotrophic and chemoorganotrophic growth were identified. Genes encoding multiple enzymes involved in anapleurotic reactions centered on C2 to C4 metabolism, including a glyoxylate bypass, were annotated. The inability of N. winogradskyi to grow on C6 molecules is consistent with the genome sequence, which lacks genes for complete Embden-Meyerhof and Entner-Doudoroff pathways, and active uptake of sugars. Two gene copies of the nitrite oxidoreductase, type I ribulose-1,5-bisphosphate carboxylase/oxygenase, cytochrome c oxidase, and gene homologs encoding an aerobic-type carbon monoxide dehydrogenase were present. Similarity of nitrite oxidoreductases to respiratory nitrate reductases was confirmed. Approximately 10% of the N. winogradskyi genome codes for genes involved in transport and secretion, including the presence of transporters for various organic-nitrogen molecules. The N. winogradskyi genome provides new insight into the phylogenetic identity and physiological capabilities of nitrite-oxidizing bacteria. The genome will serve as a model to study the cellular and molecular processes that control nitrite oxidation and its interaction with other nitrogen-cycling processes.

  19. Complete Genome Sequence of Nitrobacter hamburgensis X14 and Comparative Genomic Analysis of Species within the Genus Nitrobacter▿ †

    Science.gov (United States)

    Starkenburg, Shawn R.; Larimer, Frank W.; Stein, Lisa Y.; Klotz, Martin G.; Chain, Patrick S. G.; Sayavedra-Soto, Luis A.; Poret-Peterson, Amisha T.; Gentry, Mira E.; Arp, Daniel J.; Ward, Bess; Bottomley, Peter J.

    2008-01-01

    The alphaproteobacterium Nitrobacter hamburgensis X14 is a gram-negative facultative chemolithoautotroph that conserves energy from the oxidation of nitrite to nitrate. Sequencing and analysis of the Nitrobacter hamburgensis X14 genome revealed four replicons comprised of one chromosome (4.4 Mbp) and three plasmids (294, 188, and 121 kbp). Over 20% of the genome is composed of pseudogenes and paralogs. Whole-genome comparisons were conducted between N. hamburgensis and the finished and draft genome sequences of Nitrobacter winogradskyi and Nitrobacter sp. strain Nb-311A, respectively. Most of the plasmid-borne genes were unique to N. hamburgensis and encode a variety of functions (central metabolism, energy conservation, conjugation, and heavy metal resistance), yet ∼21 kb of a ∼28-kb “autotrophic” island on the largest plasmid was conserved in the chromosomes of Nitrobacter winogradskyi Nb-255 and Nitrobacter sp. strain Nb-311A. The N. hamburgensis chromosome also harbors many unique genes, including those for heme-copper oxidases, cytochrome b561, and putative pathways for the catabolism of aromatic, organic, and one-carbon compounds, which help verify and extend its mixotrophic potential. A Nitrobacter “subcore” genome was also constructed by removing homologs found in strains of the closest evolutionary relatives, Bradyrhizobium japonicum and Rhodopseudomonas palustris. Among the Nitrobacter subcore inventory (116 genes), copies of genes or gene clusters for nitrite oxidoreductase (NXR), cytochromes associated with a dissimilatory nitrite reductase (NirK), PII-like regulators, and polysaccharide formation were identified. Many of the subcore genes have diverged significantly from, or have origins outside, the alphaproteobacterial lineage and may indicate some of the unique genetic requirements for nitrite oxidation in Nitrobacter. PMID:18326675

  20. Genome sequence of the chemolithoautotrophic nitrite-oxidizing bacterium Nitrobacter winogradskyi Nb-255.

    Science.gov (United States)

    Starkenburg, Shawn R; Chain, Patrick S G; Sayavedra-Soto, Luis A; Hauser, Loren; Land, Miriam L; Larimer, Frank W; Malfatti, Stephanie A; Klotz, Martin G; Bottomley, Peter J; Arp, Daniel J; Hickey, William J

    2006-03-01

    The alphaproteobacterium Nitrobacter winogradskyi (ATCC 25391) is a gram-negative facultative chemolithoautotroph capable of extracting energy from the oxidation of nitrite to nitrate. Sequencing and analysis of its genome revealed a single circular chromosome of 3,402,093 bp encoding 3,143 predicted proteins. There were extensive similarities to genes in two alphaproteobacteria, Bradyrhizobium japonicum USDA110 (1,300 genes) and Rhodopseudomonas palustris CGA009 CG (815 genes). Genes encoding pathways for known modes of chemolithotrophic and chemoorganotrophic growth were identified. Genes encoding multiple enzymes involved in anapleurotic reactions centered on C2 to C4 metabolism, including a glyoxylate bypass, were annotated. The inability of N. winogradskyi to grow on C6 molecules is consistent with the genome sequence, which lacks genes for complete Embden-Meyerhof and Entner-Doudoroff pathways, and active uptake of sugars. Two gene copies of the nitrite oxidoreductase, type I ribulose-1,5-bisphosphate carboxylase/oxygenase, cytochrome c oxidase, and gene homologs encoding an aerobic-type carbon monoxide dehydrogenase were present. Similarity of nitrite oxidoreductases to respiratory nitrate reductases was confirmed. Approximately 10% of the N. winogradskyi genome codes for genes involved in transport and secretion, including the presence of transporters for various organic-nitrogen molecules. The N. winogradskyi genome provides new insight into the phylogenetic identity and physiological capabilities of nitrite-oxidizing bacteria. The genome will serve as a model to study the cellular and molecular processes that control nitrite oxidation and its interaction with other nitrogen-cycling processes.

  1. Functional role of oligomerization for bacterial and plant SWEET sugar transporter family

    Science.gov (United States)

    Xuan, Yuan Hu; Hu, Yi Bing; Chen, Li-Qing; Sosso, Davide; Ducat, Daniel C.; Hou, Bi-Huei; Frommer, Wolf B.

    2013-01-01

    Eukaryotic sugar transporters of the MFS and SWEET superfamilies consist of 12 and 7 α-helical transmembrane domains (TMs), respectively. Structural analyses indicate that MFS transporters evolved from a series of tandem duplications of an ancestral 3-TM unit. SWEETs are heptahelical proteins carrying a tandem repeat of 3-TM separated by a single TM. Here, we show that prokaryotes have ancestral SWEET homologs with only 3-TM and that the Bradyrhizobium japonicum SemiSWEET1, like Arabidopsis SWEET11, mediates sucrose transport. Eukaryotic SWEETs most likely evolved by internal duplication of the 3-TM, suggesting that SemiSWEETs form oligomers to create a functional pore. However, it remains elusive whether the 7-TM SWEETs are the functional unit or require oligomerization to form a pore sufficiently large to allow for sucrose passage. Split ubiquitin yeast two-hybrid and split GFP assays indicate that Arabidopsis SWEETs homo- and heterooligomerize. We examined mutant SWEET variants for negative dominance to test if oligomerization is necessary for function. Mutation of the conserved Y57 or G58 in SWEET1 led to loss of activity. Coexpression of the defective mutants with functional A. thaliana SWEET1 inhibited glucose transport, indicating that homooligomerization is necessary for function. Collectively, these data imply that the basic unit of SWEETs, similar to MFS sugar transporters, is a 3-TM unit and that a functional transporter contains at least four such domains. We hypothesize that the functional unit of the SWEET family of transporters possesses a structure resembling the 12-TM MFS structure, however, with a parallel orientation of the 3-TM unit. PMID:24027245

  2. Effect of mycorrhizae, Thiobacillus and sulfur nutrition on the chemical composition of soybean [Glycine max (L.)] Merr. seed.

    Science.gov (United States)

    Mostafavian, S R; Pirdashti, H; Ramzanpour, M R; Andarkhor, A A; Shahsavari, A

    2008-03-15

    A field experiment carried out in a calcareous soil with a low available phosphorus to evaluate effectiveness of biofertilizers, mycorrhizae (Glomus intraradices) and Thiobacillus sp. inoculation individually or in combination on seed yield, oil, protein and some elements (P, Fe, Mn, Zn) concentration in two soybean [Glycine max (L.) Merr.] cultivars. The applied treatments were different fertilizers with 6 levels (including: NP (control, 12 kg N ha(-1) as urea, 46 kg P2O5 ha(-1) as triple super phosphate); NPK (NP + 75 kg K2O ha(-1) as potassium sulphate); NPKS [NPK+ S (100 kg S ha(-1))]; NPKST (NPKS + seed inoculation with Thiobacillus bacteria); NPKM (NPK + Seed inoculation with mycorrhizae fungi) and NPKSTM (NPKS + seed inoculation with Thiobacillus and mycorrhizae) and two cultivars (JK and 032). Before planting, soybean seeds were inoculated by Bradyrhizobium japonicum in all treatments. Results showed that combined inoculation of biofertilizers increased yield, however the highest yield was observed in treatment NPKST. Increasing oil content (percentage) was more pronounced in treatments NPKM, while most protein content (percentage) increasing was observed in NPKS and NPKM. Fe and Zn concentrations were unaffected significantly by fertilizer treatments, but NPKSTM showed significantly higher value of seed's Mn concentration compared to treatments NP and NPK. Although no significant difference was observed in terms ofP concentration of 032 line among fertilizer treatments, JK cultivar and NPKSTM caused a significant increasing in P concentration compared to NP, NPKS and NPKM. Present results suggested that applying biofertilizers i.e., mycorrhizae and Thiobacillus increased soybean yield compared to control (NP). Overall, this study demonstrated that soybean seed yield and its chemical composition could be affected by biofertilizer inoculation.

  3. 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

  4. Complete genome sequence of Nitrobacter hamburgensis X14 and comparative genomic analysis of species within the genus Nitrobacter.

    Energy Technology Data Exchange (ETDEWEB)

    Starkenburg, Shawn R [Oregon State University; Larimer, Frank W [ORNL; Stein, Lisa Y [University of California, Riverside; Klotz, Martin G [University of Louisville, Louisville; Chain, Patrick S. G. [Lawrence Livermore National Laboratory (LLNL); Sayavedra-Soto, LA [Oregon State University; Poret-Peterson, Amisha T. [University of Louisville, Louisville; Gentry, ME [University of Louisville, Louisville; Arp, D J [Oregon State University; Ward, Bess B. [Princeton University; Bottomley, Peter J [Oregon State University

    2008-05-01

    The alphaproteobacterium Nitrobacter hamburgensis X14 is a gram-negative facultative chemolithoautotroph that conserves energy from the oxidation of nitrite to nitrate. Sequencing and analysis of the Nitrobacter hamburgensis X14 genome revealed four replicons comprised of one chromosome (4.4 Mbp) and three plasmids (294, 188, and 121 kbp). Over 20% of the genome is composed of pseudogenes and paralogs. Whole-genome comparisons were conducted between N. hamburgensis and the finished and draft genome sequences of Nitrobacter winogradskyi and Nitrobacter sp. strain Nb-311A, respectively. Most of the plasmid-borne genes were unique to N. hamburgensis and encode a variety of functions (central metabolism, energy conservation, conjugation, and heavy metal resistance), yet approximately 21 kb of a approximately 28-kb "autotrophic" island on the largest plasmid was conserved in the chromosomes of Nitrobacter winogradskyi Nb-255 and Nitrobacter sp. strain Nb-311A. The N. hamburgensis chromosome also harbors many unique genes, including those for heme-copper oxidases, cytochrome b(561), and putative pathways for the catabolism of aromatic, organic, and one-carbon compounds, which help verify and extend its mixotrophic potential. A Nitrobacter "subcore" genome was also constructed by removing homologs found in strains of the closest evolutionary relatives, Bradyrhizobium japonicum and Rhodopseudomonas palustris. Among the Nitrobacter subcore inventory (116 genes), copies of genes or gene clusters for nitrite oxidoreductase (NXR), cytochromes associated with a dissimilatory nitrite reductase (NirK), PII-like regulators, and polysaccharide formation were identified. Many of the subcore genes have diverged significantly from, or have origins outside, the alphaproteobacterial lineage and may indicate some of the unique genetic requirements for nitrite oxidation in Nitrobacter.

  5. Impact of cadmium on the bacterial communities in the gut of Metaphire posthuma

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Shih-Hsiung; Chen, Mu-Hsuan; Chen, Chien-Cheng; Chen, Colin S. [Department of Biotechnology, National Kaohsiung Normal University, Kaohsiung, Taiwan (China); Chen, Jiun-Hong [Department of Life Science, National Taiwan University, Taipei, Taiwan (China); Chen, Ssu Ching, E-mail: osycchna@ksts.seed.net.tw [Department of Biotechnology, National Kaohsiung Normal University, Kaohsiung, Taiwan (China)

    2009-12-30

    The effects of cadmium (Cd) contamination in soil onto the bacterial communities of the guts pooled from ten Metaphire posthuma were addressed during 14 days' incubation. We found that about 50% of Cd (5 mg/kg, dry weight soil) in the contaminated soil was bio-accumulated into the earthworms. DNA was extracted from the guts of M. posthuma and their dwelling soil irrespective of Cd treatment for the analysis of the bacterial communities of guts in M. posthuma and in soil by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis). A distinctive cluster of bacterial communities of the guts in the earthworm with and without Cd treatment using the analysis of unweighted pair-group method using arithmetic averages (UPGMA) was observed, indicating that the bacterial community of guts could be changed by Cd. However, no differences in the bacterial communities in soil irrespective of Cd treatment were observed, which could be resulted from the bioremediation of Cd by earthworms leading to insignificant effect of Cd on bacterial communities in soil. For the sequencing of some of the dominant bands in the DGGE profile, Bradyrhizobium japonicum, Stenotrophomonas sp. D2, and Labrys, sp. CC-BB4, whose sequences display an identity of more than 97% using blast program against a known sequence in the GeneBank database and Ribosomal database, were identified. Collectively, our results showed that earthworm treatment can decrease the concentrations of Cd in soil, and Cd cause a shift in the bacterial communities in the guts of M. posthuma. The application of M. posthuma for Cd bioremediation would be desired.

  6. Effect of soil interacting herbicides on soybean nodulation in Balcarce, Argentina

    Directory of Open Access Journals (Sweden)

    Norma Gonzalez

    1999-07-01

    Full Text Available Two trials were performed in Balcarce, Argentina (37° 45' LS; 58° 18' LW during 1993-94, to assess the effect of eight herbicides applied individually or in tank mixtures, on nodule number, nodule dry weight, seed yield and N percent in seed in soybean Asgrow 3205, inoculated with Bradyrhizobium japonicum CB 1809. Individual herbicides and doses in kg ha-1 of a.i. were metribuzin (0.48, acetochlor (0.90, metolachlor (1, flumioxazin (0.075, trifluralin (0.96, imazaquin (0.20, imazethapyr (0.10 and chlorimuron ethyl (0.0125. The mixtures were metribuzin+acetochlor (0.48+0.9, flumioxazin+acetochlor (0.075+0.9, imazaquin+acetochlor (0.2+0.9, metribuzin+metolachlor (0.48+1.92, and flumioxazin+ metolachlor (0.075+1.92. A control treatment without herbicides was included. Both trials were laid out as randomized complete blocks with four replicates, on a loam illitic thermic petrocalcic Paleudoll, 5.7% organic matter (OM, 25% clay, 30.4 cmol kg-1 CEC. Nodules were sampled at V2 (second node, V6 (sixth node and R5 (beginning seed growth stages. Herbicides did not significantly affect the beginning of nodulation or nodule number and mass at R5, not either grain yield or N accumulation. This indicates lack of interference between soil interacting herbicides and N fixation in the high organic matter, loam soils of SE Buenos Aires province, even though a tendency in less number and dry weight of nodules was evident at the two latter growth stages.

  7. A Schistosoma japonicum chimeric protein with a novel adjuvant induced a polarized Th1 immune response and protection against liver egg burdens

    Directory of Open Access Journals (Sweden)

    Xue Xiangyang

    2009-05-01

    Full Text Available Abstract Background Schitosomiasis japonica is still a significant public health problem in China. A protective vaccine for human or animal use represents an important strategy for long-term control of this disease. Due to the complex life cycle of schistosomes, different vaccine design approaches may be necessary, including polyvalent subunit vaccines. In this study, we constructed four chimeric proteins (designated SjGP-1~4 via fusion of Sj26GST and four individual paramyosin fragments. We tested these four proteins as vaccine candidates, and investigated the effect of deviating immune response on protection roles in mice. Methods The immunogencity and protection efficacy of chimeric proteins were evaluated in mice. Next, the chimeric protein SjGP-3 was selected and formulated in various adjuvants, including CFA, ISA 206, IMS 1312 and ISA 70M. The titers of antigen-specific IgG, IgE and IgG subclass were measured. The effect of adjuvant on cytokine production and percentages of CD3+CD8-IFN-γ+ cells and CD3+CD8-IL-4+ cells were analyzed at different time points. Worm burdens and liver egg counts in different adjuvant groups were counted to evaluate the protection efficacy against cercarial challenge. Results Immunization of mice with chimeric proteins provided various levels of protection. Among the four proteins, SjGP-3 induced the highest level of protection, and showed enhanced protective efficacy compared with its individual component Sj26GST. Because of this, SjGP-3 was further formulated in various adjuvants to investigate the effect of adjuvant on immune deviation. The results revealed that SjGP-3 formulated in veterinary adjuvant ISA 70M induced a lasting polarized Th1 immune response, whereas the other adjuvants, including CFA, ISA 206 and IMS 1312, generated a moderate mixed Th1/Th2 response after immunization but all except for IMS 1312 shifted to Th2 response after onset of eggs. More importantly, the SjGP-3/70M formulation induced a significant reduction in liver egg deposition at 47.0–50.3% and the number of liver eggs per female at 34.5–37.2% but less effect on worm burdens at only 17.3–23.1%, whereas no effect of the formulations with other adjuvants on the number of liver eggs per female was observed. Conclusion Construction of polyvalent subunit vaccine was capable to enhance immunogenicity and protection efficacy against schistosomiasis. There was correlation of the polarized Th1 response with reduction of liver egg burdens, supporting the immune deviation strategy for schistosomiasis japonica vaccine development.

  8. The Experimental Pathology of the Lake Lindu Strain of ’Schistosoma Japonicum’ in the Crab-Eating Macaque (Macaca Fascicularis) in Indonesia

    Science.gov (United States)

    1978-10-31

    34~ occordonce with the Principle. of tLaboatory Animal Care ettablish" by the Committee mi the Guide For Laborolry Ani-. maol Ptowurc~o, Notional Acaom ~y of...the monkey. The water bearing the crrloo was allowed to dry on the abomn of the monkey, and he wae allowed to recover In his cap . Animal A weIghd 4.5...klog1rns and was exposed to 2,005 ceroarloe per-cut•nsously on ApIl 30, 1976, while Animal B, weIghing 5.5 kilogram was exposed to 501 cercorli. per

  9. Conformational stability of pGEX-expressed Schistosoma japonicum glutathione S-transferase: a detoxification enzyme and fusion-protein affinity tag

    National Research Council Canada - National Science Library

    Kaplan, W; Hüsler, P; Klump, H; Erhardt, J; Sluis-Cremer, N; Dirr, H

    1997-01-01

    .... Size-exclusion HPLC (SEC-HPLC) and SDS-PAGE studies indicate that purification of the homodimeric protein under nonreducing conditions results in the reversible formation of significant amounts of 160-kDa and larger aggregates...

  10. Estimating the intensity of infection with Schistosoma japonicum in villagers of leyte, Philippines. Part I: a Bayesian cumulative logit model. The schistosomiasis transmission and ecology project (STEP).

    Science.gov (United States)

    Carabin, Hélène; Marshall, Clare M; Joseph, Lawrence; Riley, Steven; Olveda, Remigio; McGarvey, Stephen T

    2005-06-01

    Intensity profiles for helminths are used to describe population infection status, monitor effectiveness of control programs, and provide accurate data to validate transmission models. This