WorldWideScience

Sample records for bradyrhizobium japonicum infection1cwoa

  1. Bradyrhizobium japonicum Survival in and Soybean Inoculation with Fluid Gels.

    Science.gov (United States)

    Jawson, M D; Franzluebbers, A J; Berg, R K

    1989-03-01

    The utilization of gels, which are used for fluid drilling of seeds, as carriers of Bradyrhizobium japonicum for soybean (Glycine max (L.) Merr.) inoculation was studied. Gels of various chemical composition (magnesium silicate, potassium acrylate-acrylamide, grafted starch, and hydroxyethyl cellulose) were used, although the hydroxyethyl cellulose gels were more extensively investigated. Gel inocula were prepared by mixing gel powder with liquid cultures of B. japonicum (2% [wt/vol]). The population of B. japonicum USDA 110 did not change in each gel type during 8 days of incubation at 28 degrees C. These fluid gels were prepared with late-exponential-growth-phase cells that were washed and suspended in physiological saline. Mid-exponential-growth-phase B. japonicum USDA 110, 123, and 138 grew in cellulose gels prepared with yeast extract-mannitol broth as well as or better than in yeast extract-mannitol broth alone for the first 10 days at 28 degrees C. Populations in these cellulose gels after 35 days were as large as when the gels had originally been prepared, and survival occurred for at least 70 days. Soybeans grown in sand in the greenhouse had greater nodule numbers, nodule weights, and top weights with gel inoculants compared with a peat inoculant. In soil containing 10 indigenous B. japonicum per g of soil, inoculation resulted in increased soybean nodule numbers, nodule weights, and top weights, but only nodule numbers were greater with gel than with peat inoculation. The gel-treated seeds carried 10 to 10 more bacteria per seed (10 to 10) than did the peat-treated seeds. PMID:16347870

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

    Directory of Open Access Journals (Sweden)

    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.

  3. The Use of Acid-Aluminium Tolerant Bradyrhizobium japonicum Inoculant for Soybean Grown on Acid Soils

    Directory of Open Access Journals (Sweden)

    ANGELIA REZTY FITRIANI SITUMORANG

    2009-12-01

    Full Text Available Land with low pH soil spread widely in Indonesia can be used for soybean (Glycine max cultivation, although the production is low. The use of acid tolerant soybean and acid-Al tolerant nitrogen-fixing bacteria was an alternative way to increase soybean productivity on acid soils. This research was conducted to study the influence of acid-Al tolerant Bradyrhizobium japonicum on growth of Slamet cultivar soybean planted on acid soils in greenhouse. Three strains of acid-Al tolerant B. japonicum, i.e. BJ 11 (19, BJ 11 (5, and BJ 11 (wt, were used in this experiment. The result showed that inoculation of all acid-Al tolerant B. japonicum strains could increase plant height, shoot and root weight, number of flowers, pods, seeds, seeds dry weight, and shoot and seed nitrogen content.

  4. A novel membrane-bound glucosyltransferase from Bradyrhizobium japonicum.

    OpenAIRE

    Cohen, J. L.; Miller, K. J.

    1991-01-01

    Bacteria within the family Rhizobiaceae are distinguished by their ability to infect higher plants. The cell envelope carbohydrates of these bacteria are believed to be involved in the plant infection process. One class of cell envelope carbohydrate, the cyclic beta-1,2-glucans, is synthesized by species within two genera of this family, Agrobacterium and Rhizobium. In contrast, species of the genus Bradyrhizobium, a third genus within this family, appear to lack the capacity for cyclic beta-...

  5. Osmotic stress tolerance, PGP traits and RAPD analysis of Bradyrhizobium japonicum strains

    Directory of Open Access Journals (Sweden)

    Marinković Jelena

    2013-01-01

    Full Text Available The osmotic stress tolerance of B. japonicum strains assessed according to their persistence in PEG solution. The lowest tolerance to osmotic stress was observed in strain 511 (43.3%, and the highest tolerance was observed for strain D216 (3.3% growth reduction in presence of PEG. PGP traits of B. japonicum strains were tested. None of five B. japonicum strains produced siderophore, strains 511 and 518 had the urease ability, and only B. japonicum 518 strain showed the ability to solubilize insoluble tricalcium phosphate. RAPD analysis, using AP10, BC318, AF14 and SPH1 primers, indicated genetic differences between Bradyrhizobium strains. The first group (strains 3, 6 and 518 showed more than 80% similarity. Strains 511 and D216 formed separate clusters. Difference between strains D216 and the other strains were more than 60%, with maximum value of 72% in comparison with strain 511. Plant-growth promoting (PGP traits, osmotic stress tolerance and RAPD analysis highlighted strain D216 as useful for further investigation of B. japonicum impact on drought reduction in symbiosis with soybean. [Projekat Ministarstva nauke Republike Srbije, br. TR 31022 i br. TR 31072

  6. [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.

  7. 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...... an oxygen-consuming opening of the indole ring analogous to the one catalyzed by tryptophan 2,3-dioxygenase. The pattern of metabolite usage by known tryptophan-auxotrophic mutants and studies of metabolites by high-performance liquid chromatography indicate that anthranilic acid is a terminal degradation...

  8. 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 chromatogr...

  9. Strain selection for improvement of Bradyrhizobium japonicum competitiveness for nodulation of soybean.

    Science.gov (United States)

    Althabegoiti, María Julia; López-García, Silvina L; Piccinetti, Carlos; Mongiardini, Elías J; Pérez-Giménez, Julieta; Quelas, Juan Ignacio; Perticari, Alejandro; Lodeiro, Aníbal R

    2008-05-01

    A Bradyrhizobium japonicum USDA 110-derived strain able to produce wider halos in soft-agar medium than its parental strain was obtained by recurrent selection. It was more chemotactic than the wild type towards mannitol and three amino acids. When cultured in minimal medium with mannitol as a single carbon-source, it had one thick subpolar flagellum as the wild type, plus several other flagella that were thinner and sinusoidal. Root adsorption and infectivity in liquid media were 50-100% higher for the selected strain, but root colonization in water-unsaturated vermiculite was similar to the wild type. A field experiment was then carried out in a soil with a naturalized population of 1.8 x 10(5) soybean-nodulating rhizobia g of soil(-1). Bradyrhizobium japonicum strains were inoculated either on the soybean seeds or in the sowing furrows. Nodule occupation was doubled when the strains were inoculated in the sowing furrows with respect to seed inoculation (significant with P<0.05). On comparing strains, nodule occupation with seed inoculation was 6% or 10% for the wild type or selected strains, respectively, without a statistically significant difference, while when inoculated in the sowing furrows, nodule occupation increased to 12% and 22%, respectively (differences significant with P<0.05). PMID:18336548

  10. Role of Bradyrhizobium japonicum induced by genistein on soybean stressed by water deficit

    Energy Technology Data Exchange (ETDEWEB)

    Napoles, M. C.; Guevara, E.; Montero, F.; Roosi, A.; Ferreira, A.

    2009-07-01

    Abstract The soybean (Glycine max (L.) Merr.) is a crop mainly grown under rain fed conditions although irrigation is increasingly being used. Water deficiency is the main factor limiting seed production. The symbiosis process is also negatively affected by water stress. The isoflavone genistein have been recognized as a powerful inducer of Nod factors production by Bradyrhizobium and its addition to inocula has been shown to increase nodule number and promote soybean nitrogen (N) fixation at low temperatures. This study looks for answers about the possible role of genistein in countering the stress on nodulation produced by water deficit in soybeans. Bradyrhizobium japonicum SEMIA 5079 was grown in culture media induced or not induced with genistein. Inocula were applied to plants growing at different moisture levels. The effect of the treatments on nodulation and N content was evaluated. An improved response to drought stress was observed when the bacteria were grown in presence of genistein as a Nod factors inducer. Nodulation values under moisture stress differed from 8.9 nodules plant{sup -}1 with genistein at 10 {mu}M to 1.8 nodules plant{sup -}1 when no inducer was used. Genistein reduced the negative effect on nodulation caused by water deficiency. (Author) 43 refs.

  11. The hmuQ and hmuD Genes from Bradyrhizobium japonicum Encode Heme-Degrading Enzymes

    OpenAIRE

    Puri, Sumant; O'Brian, Mark R.

    2006-01-01

    Utilization of heme by bacteria as a nutritional iron source involves the transport of exogenous heme, followed by cleavage of the heme macrocycle to release iron. Bradyrhizobium japonicum can use heme as an iron source, but no heme-degrading oxygenase has been described. Here, bioinformatics analyses of the B. japonicum genome identified two paralogous genes renamed hmuQ (bll7075) and hmuD (bll7423) that encode proteins with weak similarity to the heme-degrading monooxygenase IsdG from Staph...

  12. Behavior of Bradyrhizobium japonicum strains under different herbicide concentrations Comportamento de estirpes de Bradyrhizobium japonicum em diferentes concentrações de herbicidas

    Directory of Open Access Journals (Sweden)

    J.S. Arruda

    2001-04-01

    Full Text Available The Bradyrhizobium japonicun strains SEMIA 5073, SEMIA 5074, SEMIA 5079 and SEMIA 5080 were grown in vitro using Vincent medium combined with different rates of the herbicides imazaquin (0, 0.04, 0.12, 0.24, 0.36 mg a.i. g-1, clomazone (0, 0.4, 0.8, 1.6 and 3.2 mg a.i. g-1 and sulfentrazone (0, 0.2, 0.4, 0.8 and 1.6 mg a.i. g-1 to evaluate the strains tolerance to herbicides. The three herbicides drastically inhibited all the rhizobium strains tested, showing a significant decrease of the CFU number as a function of herbicide rates. The rhizobium strains presented a differentiated tolerance to the herbicides. The herbicide rates that reduced 50% (I50 of the growth or survival of the rhizobium strains were below the recommended sprayed rates for weed control in the soybean crop, for all the three herbicides studied; however, sulfentrazone I50 was smaller than imazaquin and clomazone I50.As estirpes de Bradyrhizobium japonicum SEMIA 5073, SEMIA 5074, SEMIA 5079 e SEMIA 5080 foram cultivadas in vitro, utilizando meio de Vincent acrescido de diferentes doses dos herbicidas imazaquin (0; 0,04; 0,12; 0,24; 0,36 mg i.a. g-1, clomazone (0; 0,4; 0,8; 1,6; 3,2 m i.a. g-1 e sulfentrazone (0; 0,2; 0,4; 0,8; 1,6 mg i.a. g-1, com o objetivo de avaliar a tolerância destas estirpes aos herbicidas. Os três herbicidas afetaram drasticamente as estirpes de rizóbio testadas, causando decréscimo significativo no crescimento ou na sobrevivência, em função do incremento nas doses. As estirpes de rizóbio apresentaram tolerância diferenciada aos herbicidas. As doses de herbicidas que reduziram em 50% o crescimento ou a sobrevivência das estirpes (I50 foram inferiores àquelas recomendadas para o controle de plantas daninhas na cultura de soja, para os três herbicidas estudados; no entanto, o I50 do herbicida sulfentrazone foi significativamente menor que o I50 dos herbicidas imazaquin e clomazone.

  13. [Iron regulation of gene expression in the Bradyrhizobium japonicum/soybean symbiosis]. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    Guerinot, M.L.

    1992-06-01

    We wish to address the question of whether iron plays a regulatory role in the Bradyrhizobium japonicum/soybeam symbiosis. Iron may be an important regulatory signal in planta as the bacteria must acquire iron from their plant hosts and iron-containing proteins figure prominently in all nitrogen-fixing symbioses. For example, the bacterial partner is believed to synthesize the heme moiety of leghemoglobin, which may represent as much as 25--30% of the total soluble protein in an infected plant cell. For this reason, we have focused our attention on the regulation by iron of the first step in the bacterial heme biosynthetic pathway. The enzyme which catalyzes this step, 5-aminolevulinic acid synthase, is encoded by the hemA gene which we had previously cloned and sequenced. Specific objectives include: to define the cis-acting sequences which confer iron regulation on the B. japonicum hemA gene; to identify trans-acting factors which regulate the expression of hemA by iron; to identify new loci which are transcriptionally responsive to changes in iron availability; and to examine the effects of mutations in various known regulatory genes for their effect on the expression of hemA.

  14. (Iron regulation of gene expression in the Bradyrhizobium japonicum/soybean symbiosis)

    Energy Technology Data Exchange (ETDEWEB)

    Guerinot, M.L.

    1992-01-01

    We wish to address the question of whether iron plays a regulatory role in the Bradyrhizobium japonicum/soybeam symbiosis. Iron may be an important regulatory signal in planta as the bacteria must acquire iron from their plant hosts and iron-containing proteins figure prominently in all nitrogen-fixing symbioses. For example, the bacterial partner is believed to synthesize the heme moiety of leghemoglobin, which may represent as much as 25--30% of the total soluble protein in an infected plant cell. For this reason, we have focused our attention on the regulation by iron of the first step in the bacterial heme biosynthetic pathway. The enzyme which catalyzes this step, 5-aminolevulinic acid synthase, is encoded by the hemA gene which we had previously cloned and sequenced. Specific objectives include: to define the cis-acting sequences which confer iron regulation on the B. japonicum hemA gene; to identify trans-acting factors which regulate the expression of hemA by iron; to identify new loci which are transcriptionally responsive to changes in iron availability; and to examine the effects of mutations in various known regulatory genes for their effect on the expression of hemA.

  15. Mitigation of nitrous oxide emissions from soils by Bradyrhizobium japonicum inoculation

    Science.gov (United States)

    Itakura, Manabu; Uchida, Yoshitaka; Akiyama, Hiroko; Hoshino, Yuko Takada; Shimomura, Yumi; Morimoto, Sho; Tago, Kanako; Wang, Yong; Hayakawa, Chihiro; Uetake, Yusuke; Sánchez, Cristina; Eda, Shima; Hayatsu, Masahito; Minamisawa, Kiwamu

    2013-03-01

    Nitrous oxide (N2O) is a greenhouse gas that is also capable of destroying the ozone layer. Agricultural soil is the largest source of N2O (ref. ). Soybean is a globally important leguminous crop, and hosts symbiotic nitrogen-fixing soil bacteria (rhizobia) that can also produce N2O (ref. ). In agricultural soil, N2O is emitted from fertilizer and soil nitrogen. In soybean ecosystems, N2O is also emitted from the degradation of the root nodules. Organic nitrogen inside the nodules is mineralized to NH4+, followed by nitrification and denitrification that produce N2O. N2O is then emitted into the atmosphere or is further reduced to N2 by N2O reductase (N2OR), which is encoded by the nosZ gene. Pure culture and vermiculite pot experiments showed lower N2O emission by nosZ+ strains and nosZ++ strains (mutants with increased N2OR activity) of Bradyrhizobium japonicum than by nosZ- strains. A pot experiment using soil confirmed these results. Although enhancing N2OR activity has been suggested as a N2O mitigation option, this has never been tested in the field. Here, we show that post-harvest N2O emission from soybean ecosystems due to degradation of nodules can be mitigated by inoculation of nosZ+ and non-genetically modified organism nosZ++ strains of B. japonicum at a field scale.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Comparación entre coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense e inoculación simple con Bradyrhizobium japonicum en la nodulación, crecimiento y acumulación de N en el cultivo de soja Comparison of coinoculation with Bradyrhizobium japonicum and Azospirillum brasilense with inoculation with Bradyrhizobium japonicum on nodulation, growth, and N accumulation in the soybean crop

    Directory of Open Access Journals (Sweden)

    S. Benintende

    2010-12-01

    Full Text Available El objetivo fue comparar la coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense con relación a la inoculación simple con Bradyrhizobium japonicum en la nodulación, crecimiento y acumulación de N en el cultivo de soja y su rendimiento. Se realizaron dos experimentos en lotes en los que nunca se cultivó soja en años con diferentes condiciones hídricas. Los tratamientos fueron: testigo (sin inoculación, coinoculación con Bradyrhizobium japonicum y Azospirillum brasilense e inoculación simple con Bradyrhizobium japonicum. La nodulación en ambos experimentos fue superior en la coinoculación, aunque hubo diferencias significativas en el experimento con déficit hídrico. Los rendimientos, en ambos experimentos, presentaron valores medios superiores en la coinoculación, aunque sin diferencias significativas. La acumulación de biomasa aérea y N fue mayor en el año húmedo. El porcentaje de N derivado de la fijación biológica de N en el cultivo en el año seco fue de 26% para la inoculación simple y de 30% en la coinoculación, mientras que en el año húmedo este porcentaje fue de 41 y 47% respectivamente. A partir de estos resultados se concluye que existe un efecto de estimulación del crecimiento del cultivo de soja por la coinoculación. Sin embargo, es necesario continuar con estudios sobre este tema.The aim of this study was to compare coinoculation with Bradyrhizobium japonicum and Azospirillum brasilense with inoculation with Bradyrhizobium japonicum on nodulation, growth, and N accumulation in the soybean crop in field conditions. Two field tests were carried out for two years with different hydric conditions in areas not previously cultivated with soybean. The treatments were: control, coinoculation using Bradyrhizobium japonicum + Azospirillum brasilense and inoculation with Bradyrhizobium japonicum. Nodulation in both experiments was increased by dual inoculation, although significant differences were

  18. Lack of glyphosate resistance gene transfer from Roundup Ready soybean to Bradyrhizobium japonicum under field and laboratory conditions.

    Science.gov (United States)

    Isaza, Laura Arango; Opelt, Katja; Wagner, Tobias; Mattes, Elke; Bieber, Evi; Hatley, Elwood O; Roth, Greg; Sanjuán, Juan; Fischer, Hans-Martin; Sandermann, Heinrich; Hartmann, Anton; Ernst, Dieter

    2011-01-01

    A field study was conducted at the Russell E. Larson Agricultural Research Center to determine the effect of transgenic glyphosate-resistant soybean in combination with herbicide (Roundup) application on its endosymbiont Bradyrhizobium japonicum. DNA of bacteroids from isolated nodules was analysed for the presence of the transgenic 5-enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS) DNA sequence using polymerase chain reaction (PCR). To further assess the likelihood that the EPSPS gene may be transferred from the Roundup Ready (RR) soybean to B. japonicum, we have examined the natural transformation efficiency of B. japonicum strain 110spc4. Analyses of nodules showed the presence of the transgenic EPSPS DNA sequence. In bacteroids that were isolated from nodules of transgenic soybean plants and then cultivated in the presence of glyphosate this sequence could not be detected. This indicates that no stable horizontal gene transfer (HGT) of the EPSPS gene had occurred under field conditions. Under laboratory conditions, no natural transformation was detected in B. japonicum strain 110spc4 in the presence of various amounts of recombinant plasmid DNA. Our results indicate that no natural competence state exists in B. japonicum 110spc4. Results from field and laboratory studies indicate the lack of functional transfer of the CP4-EPSPS gene from glyphosate-tolerant soybean treated with glyphosate to root-associated B. japonicum.

  19. 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.

  20. An integrated biochemical system for nitrate assimilation and nitric oxide detoxification in Bradyrhizobium japonicum.

    Science.gov (United States)

    Cabrera, Juan J; Salas, Ana; Torres, María J; Bedmar, Eulogio J; Richardson, David J; Gates, Andrew J; Delgado, María J

    2016-02-01

    Rhizobia are recognized to establish N2-fixing symbiotic interactions with legume plants. Bradyrhizobium japonicum, the symbiont of soybeans, can denitrify and grow under free-living conditions with nitrate (NO3 (-)) or nitrite (NO2 (-)) as sole nitrogen source. Unlike related bacteria that assimilate NO3 (-), genes encoding the assimilatory NO3 (-) reductase (nasC) and NO2 (-) reductase (nirA) in B. japonicum are located at distinct chromosomal loci. The nasC gene is located with genes encoding an ABC-type NO3 (-) transporter, a major facilitator family NO3 (-)/NO2 (-) transporter (NarK), flavoprotein (Flp) and single-domain haemoglobin (termed Bjgb). However, nirA clusters with genes for a NO3 (-)/NO2 (-)-responsive regulator (NasS-NasT). In the present study, we demonstrate NasC and NirA are both key for NO3 (-) assimilation and that growth with NO3 (-), but not NO2 (-) requires flp, implying Flp may function as electron donor to NasC. In addition, bjgb and flp encode a nitric oxide (NO) detoxification system that functions to mitigate cytotoxic NO formed as a by-product of NO3 (-) assimilation. Additional experiments reveal NasT is required for NO3 (-)-responsive expression of the narK-bjgb-flp-nasC transcriptional unit and the nirA gene and that NasS is also involved in the regulatory control of this novel bipartite assimilatory NO3 (-)/NO2 (-) reductase pathway. PMID:26564204

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

    Science.gov (United States)

    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.

  2. DNA Microarray-Based Identification of Genes Regulated by NtrC in Bradyrhizobium japonicum.

    Science.gov (United States)

    Franck, William L; Qiu, Jing; Lee, Hae-In; Chang, Woo-Suk; Stacey, Gary

    2015-08-15

    The Bradyrhizobium japonicum NtrBC two-component system is a critical regulator of cellular nitrogen metabolism, including the acquisition and catabolism of nitrogenous compounds. To better define the roles of this system, genome-wide transcriptional profiling was performed to identify the NtrC regulon during the response to nitrogen limitation. Upon cells perceiving low intracellular nitrogen, they stimulate the phosphorylation of NtrC, which induces genes responsible for alteration of the core glutamine synthetase/glutamate synthase nitrogen assimilation pathway, including the genes for the glutamine synthetases and PII proteins. In addition, genes responsible for the import and utilization of multiple nitrogen sources, specifically nitrate and nitrite, were upregulated by NtrC activation. Mutational analysis of a candidate nitrite reductase revealed a role for NtrC in regulating the assimilation of nitrite, since mutations in both ntrC and the gene encoding the candidate nitrite reductase abolished the ability to grow on nitrite as a sole nitrogen source. PMID:26025905

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

    Directory of Open Access Journals (Sweden)

    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.

  4. 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.

  5. Inoculation with Bradyrhizobium japonicum enhances the organic and fatty acids content of soybean (Glycine max (L.) Merrill) seeds.

    Science.gov (United States)

    Silva, Luís R; Pereira, Maria J; Azevedo, Jessica; Mulas, Rebeca; Velazquez, Encarna; González-Andrés, Fernando; Valentão, Patrícia; Andrade, Paula B

    2013-12-15

    Soybean (Glycine max (L.) Merrill) is one of the most important food crops for human and animal consumption, providing oil and protein at relatively low cost. The least expensive source of nitrogen for soybean is the biological fixation of atmospheric nitrogen by the symbiotic association with soil bacteria, belonging mainly to the genus Bradyrhizobium. This study was conducted to assess the effect of the inoculation of G. max with Bradyrhizobium japonicum on the metabolite profile and antioxidant potential of its seeds. Phenolic compounds, sterols, triterpenes, organic acids, fatty acids and volatiles profiles were characterised by different chromatographic techniques. The antioxidant activity was evaluated against DPPH, superoxide and nitric oxide radicals. Inoculation with B. japonicum induced changes in the profiles of primary and secondary metabolites of G. max seeds, without affecting their antioxidant capacity. The increase of organic and fatty acids and volatiles suggest a positive effect of the inoculation process. These findings indicate that the inoculation with nodulating B. japonicum is a beneficial agricultural practice, increasing the content of bioactive metabolites in G. max seeds owing to the establishment of symbiosis between plant and microorganism, with direct effects on seed quality. PMID:23993531

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

    Science.gov (United States)

    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. PMID:27039118

  7. 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

  8. Comparison of nitrogen fixation in some Bradyrhizobium Japonicum strains using the N-15 isotopic method

    International Nuclear Information System (INIS)

    Several studies have demonstrated the significant contribution of atmospheric N fixation to soybean nutrition and growth. The variability in N fixing is due to the several factors including rhizobial strain. Hence, this study was conducted to compare some commercial Bradyrhizobium japonicum strains for the amount and proportion of N fixation in soybean (Cv. Williams) which is most commonly cultivated in Iran. Initially, strains were tested for purity, infectiveness and symbiotic effectiveness using Leonard jars and three of the more efficient strains Helinitro, Rhizoking and Gold Coat were selected. A pot experiment (four replicated) was carried out with RCBD on soil without indigenous rhizobia having 530-ppm total nitrogen under greenhouse condition. Treatments included three inoculum of selected strains (separately) and a set of non inoculated blank. For estimating BNF by A-value method, two solutions of N-15 enriched ammonium sulfate, containing 9.616 and 2.086% n-15 atom excess, were applied in amount of 20 and 100 kg N/ha at stage V2 in inoculated pots and stages V2, R2 and R5 to reference pots. During the 4 months of growth, plants were irrigated with distilled water to maintain 80% FC. Plants were harvested at stage R6. The results showed that the effects of rhizobial strains were significantly different. Gold Coat and Rhizoking strains had higher significant results compared with Helinitro. The percentages of fixed nitrogen in plants inoculated with these strains were 92.3, 90.7 and 84.7%, respectively. On the basis of these results, it may be necessary to pre-select more compatible rhizobial strain for supplying the N-requirements of soybean. (Author)

  9. Homology modeling and docking studies of BjGL, a novel (+) gamma-lactamase from Bradyrhizobium japonicum.

    Science.gov (United States)

    Song, Dawei; Zhu, Shaozhou; Li, Xingzhou; Zheng, Guojun

    2014-02-01

    (+) Gamma-lactamases are enantioselective hydrolysis enzymes that can be used to produce optically pure (-) gamma-lactam, an important pharmaceutical intermediate for the anti-AIDS drug Abacavir. In this study, homology modeling and molecular dynamic simulation studies of a 3D homology model of BjGL, a novel (+) gamma-lactamase from Bradyrhizobium japonicum, were constructed and refined. The specific substrate (+) gamma-lactam and its enantiomer (-) gamma-lactam which can not be hydrolyzed was docked into the active site respectively, and the catalytic triad and other crucial residues that participate in the formation of the hydrophobic binding pocket, hydrogen bonds, and the oxyanion hole were identified. Furthermore, possible reasons for the high diastereoselectivity of BjGL binding with the substrate are proposed. PMID:24215997

  10. Impact of elevated CO2 on carbohydrate and ureide concentrations in soybean inoculated with different strains of Bradyrhizobium japonicum

    Energy Technology Data Exchange (ETDEWEB)

    Bertrand, Annick; Prevost, Danielle; Juge, Christine; Chalifour, Francois-P.

    2011-07-15

    Over the past years, the CO2 concentration in the atmosphere has constantly increased and it is expected to continue; this climate change will lead to important impacts on the crops' productivity. The authors undertook a study to determine soybean's response to higher CO2 concentration and different strains of Bradyrhizobium japonicum. They observed that, under elevated CO2, additional photosynthates are produced and used for nodular growth and that ureides increased significantly in leaves but not much in nodules. The different strains resulted in different nodular yield and ureides concentration resulting in diverse responses to elevated CO2 concentration: the commercial strain showed a feedback inhibition of nodule activity while the indigenous strain 12NS14 with a better nodule activity had a positive feedback reaction with soybean growth. This study demonstrated that, under higher CO2 concentration, a better soybean production could be achieve with the strain 12NS14.

  11. Cloning, Sequencing and Characterization of 3-Hydroxybuty- rate Dehydrogenase Encoding Gene (bdh A) in Bradyrhizobium japonicum USDA110 Strain

    Institute of Scientific and Technical Information of China (English)

    DAI Mei-xue; WU Bo; BAI Xue-liang; ZHANG Cheng-gang; MA Qing-sheng; Charles Trevor C

    2002-01-01

    The current study describes the molecular characterization of a clone which can restore the ability of bdhA mutant strains NGRPA2 and Rm11107 to utilize 3-hydroxybutyrate as a sole carbon source (Hbu+). This clone was screened out by complementation experiment from Bradyrhizobium japonicum USDA110 genomic library, and the presence of bdhA gene in the clone was verified by Bdh assay and Southern blot analysis. Furthermore, the entire sequence of bdhA gene was sequenced and the sequence was deposited in GenBank database under the accession number AY077581. bdhA gene comprises 789 base pairs and encodes Bdh with 262 amino acid of MW 27.59 kDa. Interposon ΩKm was inserted into the bdh A ORF at EcoR I site and the bdhA mutant was constructed in B .japonicum by homologous recombination. Plant assay result did not show obvious effects of mutation of bdhA gene on nodulation and nitrogen-fixation.

  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. Crescimento aéreo e radicular da soja em solos compactados e inoculados com estirpes de Bradyrhizobium japonicum

    Directory of Open Access Journals (Sweden)

    E. V. S. Borges

    2000-03-01

    Full Text Available Um experimento com soja, utilizando amostras de um Latossolo Vermelho-Escuro álico textura média foi realizado em casa de vegetação da Universidade Federal de Uberlândia (MG, de julho/96 a fevereiro/97, visando avaliar o comportamento de estirpes de Bradyrhizobium japonicum inoculadas e nativas do solo e efeitos de níveis de compactação subsuperficial na produção de matéria seca da parte aérea (PMSPA, de raízes no anel superior da coluna e na absorção e concentração de nitrogênio na parte aérea da soja. Os tratamentos dispostos em fatorial 2 x 3 x 4 corresponderam à brometização e não-brometização do solo, presença de três estirpes de Bradyrhizobium japonicum (nativa do solo, SEMIA 5079 e SEMIA 5080 inoculadas na semente e de quatro níveis de compactação (densidades de: 1,15; 1,30; 1,45 e 1,60 kg dm-3, aplicados com prensa hidráulica no solo do anel central de uma coluna formada pela sobreposição de três anéis de PVC de 15 cm de diâmetro. Quarenta e sete dias após a emergência, as plantas foram cortadas, secas e pesadas, e as raízes do anel superior coletadas e quantificadas. A PMSPA foi crescente com a brometização do solo e inoculação com a estirpe SEMIA 5080, dentro das densidades de até 1,30 kg dm-3. Para valores maiores do que este, as estirpes estudadas não diferiram entre si. A compactação promoveu reduções lineares na produção de matéria seca da parte aérea no solo não-brometizado e parabólico, e maior produção na densidade de 1,35 kg dm-3 no solo brometizado. A concentração de nitrogênio na parte aérea foi significativamen-te menor com a inoculação da estirpe SEMIA 5079 no solo brometizado e não diferiu entre si no solo não brometizado. Maior produção de raízes na camada superficial ocorreu na ausência de brometização do solo e de inoculação com Bradyrhizobium japonicum. Com a brometização, a maior produção ocorreu com a inoculação da estirpe SEMIA 5079. A

  14. 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.

  15. Isovaleryl-homoserine lactone, an unusual branched-chain quorum-sensing signal from the soybean symbiont Bradyrhizobium japonicum.

    Science.gov (United States)

    Lindemann, Andrea; Pessi, Gabriella; Schaefer, Amy L; Mattmann, Margrith E; Christensen, Quin H; Kessler, Aline; Hennecke, Hauke; Blackwell, Helen E; Greenberg, E Peter; Harwood, Caroline S

    2011-10-01

    Many species of Proteobacteria communicate by using LuxI-LuxR-type quorum-sensing systems that produce and detect acyl-homoserine lactone (acyl-HSL) signals. Most of the known signals are straight-chain fatty acyl-HSLs, and evidence indicates that LuxI homologs prefer fatty acid-acyl carrier protein (ACP) over fatty acyl-CoA as the acyl substrate for signal synthesis. Two related LuxI homologs, RpaI and BtaI from Rhodopseudomonas palustris and photosynthetic stem-nodulating bradyrhizobia, direct production of the aryl-HSLs p-coumaroyl-HSL and cinnamoyl-HSL, respectively. Here we report that BjaI from the soybean symbiont Bradyrhizobium japonicum USDA110 is closely related to RpaI and BtaI and catalyzes the synthesis of isovaleryl-HSL (IV-HSL), a branched-chain fatty acyl-HSL. We show that IV-HSL induces expression of bjaI, and in this way IV-HSL functions like many other acyl-HSL quorum-sensing signals. Purified histidine-tagged BjaI was an IV-HSL synthase, which was active with isovaleryl-CoA but not detectably so with isovaleryl-ACP. This suggests that the RpaI-BtaI-BjaI subfamily of acyl-HSL synthases may use CoA- rather than ACP-linked substrates for acyl-HSL synthesis. The bjaI-linked bjaR(1) gene is involved in the response to IV-HSL, and BjaR(1) is sensitive to IV-HSL at concentrations as low as 10 pM. Low but sufficient levels of IV-HSL (about 5 nM) accumulate in B. japonicum culture fluid. The low levels of IV-HSL synthesis have likely contributed to the fact that the quorum-sensing signal from this bacterium has not been described elsewhere.

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

    OpenAIRE

    Nisha Kadian; Kuldeep Yadav; Ashok Aggarwal

    2014-01-01

    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 effec...

  17. An L-glucitol oxidizing dehydrogenase from Bradyrhizobium japonicum USDA 110 for production of D-sorbose with enzymatic or electrochemical cofactor regeneration.

    Science.gov (United States)

    Gauer, Sabrina; Wang, Zhijie; Otten, Harm; Etienne, Mathieu; Bjerrum, Morten Jannik; Lo Leggio, Leila; Walcarius, Alain; Giffhorn, Friedrich; Kohring, Gert-Wieland

    2014-04-01

    A gene in Bradyrhizobium japonicum USDA 110, annotated as a ribitol dehydrogenase (RDH), had 87 % sequence identity (97 % positives) to the N-terminal 31 amino acids of an L-glucitol dehydrogenase from Stenotrophomonas maltophilia DSMZ 14322. The 729-bp long RDH gene coded for a protein consisting of 242 amino acids with a molecular mass of 26.1 kDa. The heterologously expressed protein not only exhibited the main enantio selective activity with D-glucitol oxidation to D-fructose but also converted L-glucitol to D-sorbose with enzymatic cofactor regeneration and a yield of 90 %. The temperature stability and the apparent K m value for L-glucitol oxidation let the enzyme appear as a promising subject for further improvement by enzyme evolution. We propose to rename the enzyme from the annotated RDH gene (locus tag bll6662) from B. japonicum USDA as a D-sorbitol dehydrogenase (EC 1.1.1.14).

  18. EFFECT OF FERULIC ACID ON CHEMOTAXIS AND NODULATION OF Bradyrhizobium japonicum

    Directory of Open Access Journals (Sweden)

    María C. Nápoles

    2001-01-01

    Full Text Available En este trabajo se estudió el efecto de tres concentraciones de ácido ferúlico sobre la quimiotaxis y la nodulación de B. japonicum ICA 8001. También se evaluó el efecto de este ácido hidroxicinámico obtenido a partir de vainillina, así como su capacidad de inducción sobre los genes de nodulación mediante la detección de factores Nod sintetizados. Se obtuvo una actividad quimiotáctica positiva pero no fuerte y solamente 10 mM como componente del medio de cultivo mostró una influencia positiva sobre la nodulación. El ácido ferúlico sintetizado a partir de vainillina incrementó todos los parámetros de la nodulación. La actividad nod inductora de este ácido se evidenció con la producción de cuatro estructuras de lipoquitinoligosacáridos.

  19. Functional Bradyrhizobium japonicum NifA expression under a hybrid nptII-nifH promoter in E. coli and Acetobacter diazotrophicus SRT4.

    Science.gov (United States)

    Menéndez, C; Selman-Housein, G; Arrieta, J G; Coego, A; Hernández, L

    1998-01-01

    A hybrid promoter consisting of the in tandem fusion of the Tn5 nptII and the Klebsiella pneumoniae nifH promoters was constructed to study the functionality of the nif genes transcriptional activator NifA from Bradyrhizobium japonicum in two different host bacteria. beta-galactosidase experiments in E. coli revealed that the hybrid nptII-nifH promoter can behave as a constitutive or a NifA-inducible promoter depending on the aeration conditions. Expression of the B. japonicum NifA from the hybrid nptII-nifH promoter (plasmid pBPF204) induced "in trans" lacZ transcription from the Azotobacter chroococcum nifH promoter in E. coli and A. diazotrophicus cells grown at low pO2. Similarly, the plasmid pBPF204 increased nitrogenase activity in A. diazotrophicus cells grown under microaerobic conditions. Based on these results, we suggest that the B. japonicum NifA could function as an efficient O2-sensitive transcriptional activator of nif genes in genetically distant diazotrophic bacteria. PMID:10932742

  20. 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.

  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. Expression of animal anti-apoptotic gene Ced-9 enhances tolerance during Glycine max L.-Bradyrhizobium japonicum interaction under saline stress but reduces nodule formation.

    Directory of Open Access Journals (Sweden)

    Germán Robert

    Full Text Available 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.

  3. The Bradyrhizobium japonicum Ferrous Iron Transporter FeoAB Is Required for Ferric Iron Utilization in Free Living Aerobic Cells and for Symbiosis.

    Science.gov (United States)

    Sankari, Siva; O'Brian, Mark R

    2016-07-22

    The bacterium Bradyrhizobium japonicum USDA110 does not synthesize siderophores for iron utilization in aerobic environments, and the mechanism of iron uptake within symbiotic soybean root nodules is unknown. An mbfA bfr double mutant defective in iron export and storage activities cannot grow aerobically in very high iron medium. Here, we found that this phenotype was suppressed by loss of function mutations in the feoAB operon encoding ferrous (Fe(2+)) iron uptake proteins. Expression of the feoAB operon genes was elevated under iron limitation, but mutants defective in either gene were unable to grow aerobically over a wide external ferric (Fe(3+)) iron (FeCl3) concentration range. Thus, FeoAB accommodates iron acquisition under iron limited and iron replete conditions. Incorporation of radiolabel from either (55)Fe(2+) or (59)Fe(3+) into cells was severely defective in the feoA and feoB strains, suggesting Fe(3+) reduction to Fe(2+) prior to traversal across the cytoplasmic membrane by FeoAB. The feoA or feoB deletion strains elicited small, ineffective nodules on soybean roots, containing few bacteria and lacking nitrogen fixation activity. A feoA(E40K) mutant contained partial iron uptake activity in culture that supported normal growth and established an effective symbiosis. The feoA(E40K) strain had partial iron uptake activity in situ within nodules and in isolated cells, indicating that FeoAB is the iron transporter in symbiosis. We conclude that FeoAB supports iron acquisition under limited conditions of soil and in the iron-rich environment of a symbiotic nodule. PMID:27288412

  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. Enhanced Survival and Nodule Occupancy of Pigeon pea Nodulating Rhizobium sp. ST1 expressing fegA Gene of Bradyrhizobium japonicum 61A152

    Directory of Open Access Journals (Sweden)

    G. Archana

    2009-01-01

    Full Text Available Problem statement: Rhizobial isolates belonging to genera (Rhizobium sp. and Mesorhizobium sp. in our laboratory produced only catecholate type of siderophores. Although FhuA and FegA (ferrichrome receptors homologs were found to be present in the sequenced genomes of few rhizobia (e.g., 1 in R. etli and 2 in Mesorhizobium sp. BNC1, laboratory isolates of the corresponding genera failed to utilize ferrichrome, a siderophore which is present in nanomolar concentrations in the soil. This inability was considered as a negative fitness factor with respect to rhizospheric colonization by these rhizobia. Approach: The 2.4 kb fegA gene (encoding ferrichrome receptor was amplified along with its native promoter from Bradyrhizobium japonicum 61A152 and cloned in a broad host range plasmid vector pUCPM18. The plasmid construct pFJ was transferred by conjugation into Rhizobium sp. ST1 to give transconjugant ST1pFJ12. The consequence of FegA expression on the transconjugant was tested under lab and soil conditions, using physiological experiments. Results: Ability of the transconjugant ST1pFJ12 to utilize ferrichrome and expression of a 79 kD protein band on the outer membrane of the transconjugant confirmed FegA expression. Transconjugant ST1pFJ12 exhibited increased growth rate as compared to the parent strain ST1, in minimal media containing ferrichrome as the sole iron source, confirming the positive effect of FegA expression. Inoculation of pigeon pea seedlings with transconjugant ST1pFJ12 led to a marked increase in plant growth parameters as compared to plants inoculated with the parent strain ST1, the effect being more pronounced when Ustilago maydis, a ferrichrome producer was co-inoculated in the systems. Nodule occupancy on pigeon pea plant when inoculated with the transconjugant ST1pFJ12 alone was 57% which increased to 66% when co-inoculated with U. maydis as compared with 37 and 30

  6. Relation between soybean nodulation byBradyrhizobium japonicum and intensity of infestation with the spider miteTetranychus cucurbitacearum (Saved) under different sowing dates and increasing amounts of nitrogenous fertilizer.

    Science.gov (United States)

    Yanni, Y G; Hassan, A A; El-Beheiry, M M; Hassan, M E

    1991-01-01

    Inoculation of soybeans withBradyrhizobium japonicum and also the time of sowing of soybeans were studied under N-fertilization with urea at 0, 96 and 192 kg N/ha. Intensity of infestation with the spider miteTetranychus cucurbitacearum (Saved) was followed over six weeks. Inoculation greatly enhanced nodule numbers and weight, dry weight and N content of 65-day-old plant, seed yield, seed N content, 100-seed weight and efficiency of use of N from urea. Counts of the spider mite were increased by inoculation and N fertilization but decreased sharply under late plantation. Fertilizer N proportionally decreased nodulation, N2-fixation and productivity of chemical fertilizer. A late sowing date gave the least seed production but the maximum percentage increase in seed yield if inoculated. This indicated a lower efficiency of chemical fertilizer N utilization and the necessity for inoculation.

  7. CO-INOCULAÇÃO E MODOS DE APLICAÇÃO DE Bradyrhizobium japonicum e Azospirillum brasilense E ADUBAÇÃO NITROGENADA NA NODULAÇÃO DAS PLANTAS E RENDIMENTO DA CULTURA DA SOJA

    Directory of Open Access Journals (Sweden)

    Alessandro Lucca Braccini

    2016-03-01

    Full Text Available As new cultivation techniques arise, crops require the development of new technologies and management practices that aim higher yields with lower production cost. Thus, the objective of this study was to evaluate the efficiency of co-inoculation and different ways of application of Bradyrhizobium japonicum and Azospirillum brasilense and nitrogen fertilization on nodulation and yield of soybean. The experiment was established in November 2013 in an area located in the Iguatemi Experimental Farm, of State University of Maringá (UEM. The design was a randomized block with four replications. The treatments consisted of a control, fertilization with nitrogen (200 kg N ha-1, standard inoculation in seeds (liquid and peat inoculant, co-inoculation on seed and co-inoculation at sowing, using different doses of inoculant B. japonicum and A. brasilense. The following determinations were carried out in the field and in the laboratory: number of nodes at flowering, mass of nodules at flowering, the shoot dry mass, nitrogen content in the shoot, grain nitrogen content, number of pods/plant, thousand grain mass and yield. The results showed that inoculation through seed treatment with liquid inoculant presents an adequate option to farmers, in the conditions of this experiment. Nitrogen fertilization did not increase yield and yield components, being unnecessary.

  8. Análisis de la interacción soya-cepa (bradyrhizobium japonicum) x ambiente, en oxisoles de la orinoquia colombiana

    OpenAIRE

    Valencia, Rubén; Ligarreto M., Gustavo

    2012-01-01

    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 ...

  9. An L-glucitol oxidizing dehydrogenase from Bradyrhizobium japonicum USDA 110 for production of D-sorbose with enzymatic or electrochemical cofactor regeneration

    DEFF Research Database (Denmark)

    Gauer, Sabrina; Wang, Zhijie; Otten, Harm;

    2014-01-01

    stability and the apparent K m value for L-glucitol oxidation let the enzyme appear as a promising subject for further improvement by enzyme evolution. We propose to rename the enzyme from the annotated RDH gene (locus tag bll6662) from B. japonicum USDA as a D-sorbitol dehydrogenase (EC 1.1.1.14)....

  10. EVALUACIÓN AGRONÓMICA DEL ANÁLOGO DE BRASINOESTEROIDES BB-6 EN SOYA, INOCULADA CON Bradyrhizobium japonicum Y HMA, CULTIVADA EN INVIERNO SOBRE UN SUELO FERRALSOL

    Directory of Open Access Journals (Sweden)

    J. Corbera

    2004-01-01

    Full Text Available En las condiciones de un suelo Ferralsol del Instituto Nacional de Ciencias Agrícolas, ubicado en San José de las Lajas, provincia La Habana, se desarrolló un estudio de campo con diferentes cultivares de soya en época de siembra de invierno, con el objetivo de evaluar los efectos agronómicos que sobre el cultivo inoculado con Bradyrhizobium japonicum y hongos formadores de micorriza arbuscular, produce la aplicación de un análogo de brasinoesteroides. Los resultados mostraron diferencias significativas entre tratamientos para todos los cultivares en estudio, corroborándose la respuesta de esta leguminosa a la aplicación conjunta de los productos biofertilizantes, lográndose incrementos del rendimiento entre 17.07 y 33.91 %. Se evidenció el efecto positivo del biorregulador aplicado, en el crecimiento y desarrollo del cultivo, obteniéndose los mayores incrementos de los rendimientos para el tratamiento coinoculado más la aplicación del biorregulador, con valores que oscilaron entre 29.67 y 42.61 % en dependencia del cultivar evaluado. Es de destacar, además, que la aplicación del análogo de brasinoesteroides logró rendimientos superiores a todos sus homólogos biofertilizados, con incrementos del rendimiento entre 4.24 y 10.45 %.

  11. OPTIMIZATION OF CO-CULTIVATION CONDITIONS OF Chlamydomonas reinhardtii AND Bradyrhizobium japonicum FOR HYDROGEN PRODUCTION%莱茵衣藻和根瘤菌共培养提高产氢条件的优化

    Institute of Scientific and Technical Information of China (English)

    许丽丽; 徐潇; 吴双秀; 王全喜; 李德志

    2015-01-01

    将莱茵衣藻(Chlamydomonas reinhardtii)以不同比例与日本慢生大豆根瘤菌(Bradyrhizobium japonicum)混合,在不同光照条件下进行产氢培养,以确定产氢的最优条件和探索产氢提高的机理.结果表明:藻菌共培养的最优产氢条件为25℃、光照200 μE/(m2·s)、生长至饱和期的菌和藻体积比为1:80,产氢量最大,约为272 μmol/(mg Chl),是对照组的17.0倍.藻菌共培养提高产氢量的主要原因是体系中O2浓度的降低使氢化酶活性提高以及衣藻生物量的增加.

  12. Differential control of Bradyrhizobium japonicum iron stimulon genes through variable affinity of the iron response regulator (Irr) for target gene promoters and selective loss of activator function.

    Science.gov (United States)

    Jaggavarapu, Siddharth; O'Brian, Mark R

    2014-05-01

    Bradyrhizobium japonicum Irr is a conditionally stable transcriptional activator and repressor that accumulates in cells under iron-limited, manganese-replete conditions, but degrades in a haem-dependent manner under high iron conditions, manganese limitation or upon exposure to H2 O2 . Here, we identified Irr-regulated genes that were relatively unresponsive to factors that promote Irr degradation. The promoters of those genes bound Irr with at least 200-fold greater affinity than promoters of the responsive genes, resulting in maintenance of promoter occupancy over a wide cellular Irr concentration range. For Irr-repressible genes, promoter occupancy correlated with transcriptional repression, resulting in differential levels of expression based on Irr affinity for target promoters. However, inactivation of positively controlled genes required neither promoter vacancy nor loss of DNA-binding activity by Irr. Thus, activation and repression functions of Irr may be uncoupled from each other under certain conditions. Abrogation of Irr activation function was haem-dependent, thus haem has two functionally separable roles in modulating Irr activity. The findings imply a greater complexity of control by Irr than can be achieved by conditional stability alone. We suggest that these regulatory mechanisms accommodate the differing needs for Irr regulon genes in response to the prevailing metabolic state of the cell.

  13. Respuesta diferencial de variedades de soya a la asociación simbiótica con cepas de Bradyrhizobium japonicum, en Oxisoles de la Orinoquia colombiana / Differental response of soybean varieties to the symbiotic association with Bradyrhizobium japonicum strains, in Oxisols of the Colombian Orinoquia

    OpenAIRE

    Valencia Ramírez, Rubén Alfredo

    2010-01-01

    El cultivo de la soya (Glycine max (L) Merril), en oxisoles de la Orinoquia colombiana presenta restricciones por la alta saturación de aluminio y en general, baja CIC y MO, que afectan la fijación biológica del N y el crecimiento normal de las plantas. La respuesta diferencial de variedades de soya a la inoculación con diferentes cepas de B. japonicum, se determinó mediante la evaluación de seis genotipos inoculados individualmente con las cepas ICA J-01, J-96, J-98, la mezcla de J-01 con J-...

  14. Root Nodule Bradyrhizobium spp. Harbor tfdAα and cadA, Homologous with Genes Encoding 2,4-Dichlorophenoxyacetic Acid-Degrading Proteins

    OpenAIRE

    Itoh, Kazuhito; Tashiro, Yoshiko; Uobe, Kazuko; Kamagata, Yoichi; Suyama, Kousuke; Yamamoto, Hiroki

    2004-01-01

    The distribution of tfdAα and cadA, genes encoding 2,4-dichlorophenoxyacetate (2,4-D)-degrading proteins which are characteristic of the 2,4-D-degrading Bradyrhizobium sp. isolated from pristine environments, was examined by PCR and Southern hybridization in several Bradyrhizobium strains including type strains of Bradyrhizobium japonicum USDA110 and Bradyrhizobium elkanii USDA94, in phylogenetically closely related Agromonas oligotrophica and Rhodopseudomonas palustris, and in 2,4-D-degradin...

  15. Subcloning and sequencing analysis of a 16 kb EcoRⅠ DNA fragment of the chromosome of Bradyrhizobium japonicum%慢生型大豆根瘤菌染色体的16kb EcoRⅠ DNA片段的亚克隆测序分析

    Institute of Scientific and Technical Information of China (English)

    唐咸来; 吕安国; 武波; 柏学亮; 唐东阶; 唐纪良; 马庆生

    2001-01-01

    亚克隆测序分析发现,在Bradyrhizobium japonicum的GX201菌株染色体的16kb EcoRⅠ DNA区段上含有一个与putA基因同源的基因的ORF 3054(Open Reading Frame).该基因ORF长3054bp,在核苷酸水平上与已报道的putA基因有94%一致性.其推断性的编码产物在氨基酸水平上与putA编码产物脯氨酸脱氢酶(ProDH)有95%一致性.利用Tn5gusA5诱变的方法获得了该同源基因的标记置换突变体,该突变体在液体培养基(YMA)中的生长速率比野生菌株慢.%The results of the subcloning and sequencing analysis showed that there is a perfect ORF3054 (Open Reading Frame) homologus with the putA gene in the 16kb EcoRⅠ DNA fragment cloned from the chromosome of Bradyrhizobium japonicum GX201,the length of which was 3054 base pairs.It shared 94% identity with the DNA of the putA gene at the nucleotide level,and its putative product protein showed 95% identity with the proline dehydrogenase (ProDH) encoded by putA gene at amino acid level.When cultured in the liquid medium YMA,the growth ratio of the markerexchanger mutant of this homologous gene was lower than that of the wildtype strain.

  16. Optimization of co-cultivation conditions of transgenic alga hemHc-lbac and Bradyrhizobium japonicum for hydrogen production%转基因莱茵衣藻hemHc-lbac和根瘤菌共培养提高产氢培养条件的优化

    Institute of Scientific and Technical Information of China (English)

    许丽丽; 王全喜; 吴双秀; 李德志

    2014-01-01

    转基因莱茵衣藻hemHc-lbac(transgenic Chlamydomonas reinhardtii hemHc-lbac)以不同比例与日本慢生大豆根瘤菌(Bradyrhizobium japonicum)混合,在不同光照条件下进行产氢培养,以确定产氢的最优条件和探索产氢提高的机理.结果表明藻菌共培养的最优产氢条件为25℃、光照30 μE·m-2·s-1、生长至饱和期的菌和藻体积比为1∶80,产氢量达到最大,约为278 μmol·mg-1Chl,是对照组80 μmol·mg-1Chl的3.5倍.藻菌共培养提高产氢量的主要原因是体系中氧气浓度的降低而使氢化酶活性提高、以及衣藻生物量的增加.该研究为利用藻菌共培养及转基因的方法提高微藻光合生物制氢效率提供了重要实验基础.

  17. Improvement of Nitrogen Fixation Efficiency and Plasmid Stability in Bradyrhizobium japonicum by the Introduction of dctABD and parCBA/DE Genes%导入dctABD和parCBA/DE基因提高大豆慢生根瘤菌固氮效率和稳定性的研究

    Institute of Scientific and Technical Information of China (English)

    李友国; 李杰; 刘墨青; 周俊初

    2000-01-01

    以pLAFR3为载体构建重组质粒pHN207,携带有来自苜蓿根瘤菌(Sinorhizobium meliloti)的四碳二羧酸转移酶基因dctABD、来自pTR102的parCBA/DE基因和标记发光酶基因luxAB。利用2亲本杂交法,将重组质粒pHN207导入大豆慢生根瘤菌(Bradyrhizobium japonicum)TA11和CB1809,分别考察了转移接合子中外源重组质粒在人工培养条件和共生条件下的稳定性,结果表明par基因的引入明显提高pLAFR3在TA11和CB1809中的稳定性。dctABD基因可显著提高TA11和CB1809在大豆黑龙33、宁镇一号和渝豆一号上的共生固氮能力,使结瘤植物的地上部分干重(生物量)和总氮量等指标较对照组有显著提高。%ArecombinantplasmidpHN207containingC4-dicarboxylicacidtransport genes(dctABD) from Sinorhizobium meliloti, parCBA/DE genes from pTR102 and reporter genes luxAB from pDB30 was constructed by using pLAFR3 as the vector. The pHN207 was then introduced into the Bradyrhizobium japonicum TA11 and CB1809 by bi-parental mating. It was confirmed that parCBA/DE genes could increase the stability of pLAFR3 in the transconjugants under both free-living and symbiotic condition. The results of plant pot experiment indicated that the introduction of dctABD genes could significantly improve the symbiotic nitrogen fixation efficiency of TA11 and CB1809 with soybean varieties of Heilong 33, Ningzhen No.1 and Yudou No.1. Compared with the control, the shoot dry weight (biomass) and total nitrogen content of the plants tested were significantly increased.

  18. Arsenic effect on the model crop symbiosis Bradyrhizobium-soybean.

    Science.gov (United States)

    Talano, Melina A; Cejas, Romina B; González, Paola S; Agostini, Elizabeth

    2013-02-01

    Soybean (Glycine max) is often being cultivated in soils with moderate to high arsenic (As) concentrations or under irrigation with As contaminated groundwater. The purpose of this study was to determine the effect of As on soybean germination, development and nodulation in soybean-Bradyrhizobium japonicum E109 symbiosis, as a first-step approach to evaluate the impact of As on soybean production. Semi-hydroponic assays were conducted using soybean seedlings inoculated and non-inoculated with B. japonicum E109 and treated with arsenate or arsenite. Soybean germination and development, at early stage of growth, were significantly reduced from 10 μM arsenate or arsenite. This also was seen for soybean seedlings inoculated with B. japonicum mainly with arsenite where, in addition, the number of effective nodules was reduced, despite that the microorganism tolerated the metalloid. This minor nodulation could be due to a reduced motility (swarming and swimming) of the microorganism in presence of As. Arsenic concentration in roots was about 250-times higher than in shoots. Transference coefficient values indicated that As translocation to aerial parts was low and As accumulated mainly in roots, without significant differences between inoculated and non-inoculated plants. The presence of As restricted soybean-B. japonicum symbiosis and hence, the efficiency of most used commercial inoculants for soybean. Thus, water and/or soils containing As would negatively impact on soybean production, even in plants inoculated with B. japonicum E109.

  19. Multidimensional cluster stability analysis from a Brazilian Bradyrhizobium sp. RFLP/PCR data set

    Science.gov (United States)

    Milagre, S. T.; Maciel, C. D.; Shinoda, A. A.; Hungria, M.; Almeida, J. R. B.

    2009-05-01

    The taxonomy of the N2-fixing bacteria belonging to the genus Bradyrhizobium is still poorly refined, mainly due to conflicting results obtained by the analysis of the phenotypic and genotypic properties. This paper presents an application of a method aiming at the identification of possible new clusters within a Brazilian collection of 119 Bradyrhizobium strains showing phenotypic characteristics of B. japonicum and B. elkanii. The stability was studied as a function of the number of restriction enzymes used in the RFLP-PCR analysis of three ribosomal regions with three restriction enzymes per region. The method proposed here uses clustering algorithms with distances calculated by average-linkage clustering. Introducing perturbations using sub-sampling techniques makes the stability analysis. The method showed efficacy in the grouping of the species B. japonicum and B. elkanii. Furthermore, two new clusters were clearly defined, indicating possible new species, and sub-clusters within each detected cluster.

  20. A role of Bradyrhizobium elkanii and closely related strains in the degradation of methoxychlor in soil and surface water environments.

    Science.gov (United States)

    Satsuma, Koji; Masuda, Minoru; Sato, Kiyoshi

    2013-01-01

    We have reported that a leguminous bacterial strain, Bradyrhizobium sp. strain 17-4, isolated from river sediment, phylogenetically very close to Bradyrhizobium elkanii, degraded methoxychlor through O-demethylation and oxidative dechlorination. In the present investigation, we found that B. elkanii (USDA94), a standard species deposited in the Culture Collection, degraded methoxychlor. Furthermore, Bradyrhizobium sp. strain 4-1, also very close to B. elkanii, isolated from Japanese paddy field soil, degraded methoxychlor. These B. elkanii and closely related strains degraded methoxychlor through almost identical metabolic pathways, and cleaved the phenyl ring and mineralized. In contrast, another representative Bradyrhizobium species, B. japonicum (USDA110), did not degrade methoxychlor at all. Based on these findings, B. elkanii and closely related strains are likely to play an important role not only in providing the readily biodegradable substrates but also in completely degrading (mineralizing) methoxychlor by themselves in the soil and surface water environment.

  1. 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.

  2. Bradyrhizobium strain and the 15N natural abundance quantification of biological N2 fixation in soybean

    International Nuclear Information System (INIS)

    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 15N natural abundance technique to quantify BNF inputs, it is essential to determine, with accuracy, the 15N 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 δ15N natural abundance in a non-labelled soil, and estimate of the contribution of BNF derived from the use of 15N-isotope dilution in soils enriched with 15N. To evaluate N2 fixation by soybean, three non-N2-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 N2 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 N2 fixation. (author)

  3. 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

    2012-12-01

    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

  4. Taxonomy Icon Data: Schistosoma japonicum [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Schistosoma japonicum Schistosoma japonicum Platyhelminthes Schistosoma_japonicum_L.png Schistosoma_japonic...um_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+japonic...um&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japonic...um&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 ...

  5. THE EFFECT OF INOCULATING WITH ARBUSCULAR MYCORRHIZA AND BRADYRHIZOBIUM STRAINS ON SOYBEAN (Glycine max (L Merrill CROP DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    Marlen Hernández

    2003-01-01

    Full Text Available El presente trabajo se realizó durante la primavera del 2001, en áreas de la Estación Experimental del Arroz "Los Palacios", perteneciente al Instituto Nacional de Ciencias Agrícolas. Se evaluó la respuesta de la variedad de soya G7-R-315, ante las inoculaciones simple y combinada de las cepas de Bradyrhizobium japonicum y Glomus fasciculatum. Para ello se realizó un experimento empleando un diseño de bloques al azar con seis tratamientos y cuatro réplicas. Los resultados mostraron influencia positiva de los microorganismos utilizados sobre la altura y el rendimiento de las plantas de soya, obteniéndose los mejores resultados en los tratamientos donde se combinó la inoculación de la semilla con ambas cepas, así como en el tratamiento donde se sustituyó la fertilización nitrogenada por Bradyrhizobium japonicum.

  6. 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

  7. Especificidade hospedeira de variantes Bradyrhizobium spp em soja (cvs peking e clark, caupi e guandu Host specificity of Bradyrhizobium spp mutants in soybean (cultivars peking and clark, cowpea and pigeon pea

    Directory of Open Access Journals (Sweden)

    Fabíola Gomes de Carvalho

    2008-12-01

    Full Text Available A expansão da cultura da soja evidenciou uma alta especificidade hospedeira, requerendo a pesquisa de novas estirpes que apresentassem capacidade de nodular a soja e bom potencial de competição com a população de rizóbios naturalizada nos solos. O objetivo deste trabalho foi avaliar a especificidade hospedeira de variantes isolados de estirpes de Bradyrhizobium spp quanto à nodulação e eficiência relativa de fixação de N2 atmosférico em soja (cvs peking e clark, caupi e guandu. O experimento foi realizado sob condições controladas em câmara de crescimento por meio de testes de variantes de B. japonicum e B. elkanii e suas respectivas estirpes originais quanto à habilidade de nodular soja, caupi e guandu. A colheita foi realizada aos 35 dias, sendo avaliada a nodulação (número, peso dos nódulos secos, produção de matéria seca na parte aérea, eficiência relativa de fixação de N2 atmosférico. Os variantes e estirpes de Bradyrhizobium spp nodularam Glycine max (cultivares BR-16, Clark e Peking, Vigna unguiculata e Cajanus cajan, contudo, apenas para Glycine max a interação rizóbio-leguminosa demonstrou eficiência simbiótica significativa.The expansion of the soybean crop has evidenced a high host specificity, indicating the requirement of new strains with capacity of nodulating soybean and competition capacity with the natural soil rhizobial population. The purpose of this study was to evaluate the host specificity of mutants isolated from Bradyrhizobium spp strains in relation to nodulation and relative N2 fixation effectiveness in soybean (cultivars Peking and Clark, cowpea and pigeon pea. The experiment was carried out under controlled conditions in a growth chamber, where B. japonicum and B. elkanii mutants and the respective original strains were tested for their nodulation ability on soybean, cowpea and pigeon pea. The crop was harvested 35 days after planting and the nodulation (number, dry nodule weight

  8. Small RNAs of the Bradyrhizobium/Rhodopseudomonas lineage and their analysis.

    Science.gov (United States)

    Madhugiri, Ramakanth; Pessi, Gabriella; Voss, Björn; Hahn, Julia; Sharma, Cynthia M; Reinhardt, Richard; Vogel, Jörg; Hess, Wolfgang R; Fischer, Hans-Martin; Evguenieva-Hackenberg, Elena

    2012-01-01

    Small RNAs (sRNAs) play a pivotal role in bacterial gene regulation. However, the sRNAs of the vast majority of bacteria with sequenced genomes still remain unknown since sRNA genes are usually difficult to recognize and thus not annotated. Here, expression of seven sRNAs (BjrC2a, BjrC2b, BjrC2c, BjrC68, BjrC80, BjrC174 and BjrC1505) predicted by genome comparison of Bradyrhizobium and Rhodopseudomonas members, was verified by RNA gel blot hybridization, microarray and deep sequencing analyses of RNA from the soybean symbiont Bradyrhizobium japonicum USDA 110. BjrC2a, BjrC2b and BjrC2c belong to the RNA family RF00519, while the other sRNAs are novel. For some of the sRNAs we observed expression differences between free-living bacteria and bacteroids in root nodules. The amount of BjrC1505 was decreased in nodules. By contrast, the amount of BjrC2a, BjrC68, BjrC80, BjrC174 and the previously described 6S RNA was increased in nodules, and accumulation of truncated forms of these sRNAs was observed. Comparative genomics and deep sequencing suggest that BjrC2a is an antisense RNA regulating the expression of inositol-monophosphatase. The analyzed sRNAs show a different degree of conservation in Rhizobiales, and expression of homologs of BjrC2, BjrC68, BjrC1505, and 6S RNA was confirmed in the free-living purple bacterium Rhodopseudomonas palustris 5D.

  9. Diversifying selection by Desmodiinae legume species on Bradyrhizobium symbionts.

    Science.gov (United States)

    Parker, Matthew A; Jankowiak, Jennifer G; Landrigan, Grace K

    2015-07-01

    Desmodium and Hylodesmum (Papilionoideae Subtribe Desmodiinae) are among the most common herbaceous perennial legumes native to eastern North America. To analyze the population structure of their Bradyrhizobium sp. root-nodule bacteria, 159 isolates were sampled from ten host species across a 1000 km region. Phylogenetic analysis of four housekeeping loci (2164 bp) and two loci in the symbiosis island (SI) chromosomal region (1374 bp) indicated extensive overlap in symbiont utilization, with each common bacterial clade found on 2-7 species of these legume genera. However, host species differed considerably in the relative proportion of symbionts belonging to different Bradyrhizobium clades. High phylogenetic incongruence between trees for housekeeping loci and SI loci suggested that diversification of these Bradyrhizobium lineages involved substantial horizontal gene transfer. Plant inoculation with strains from six Bradyrhizobium clades revealed marked disparity in relative bacterial reproductive success across four Desmodium species. Estimated yield of Bradyrhizobium progeny cells per plant ranged from zero to >10(9), and strains with high fitness on one host sometimes reproduced poorly on other host species. Diversifying selection on bacteria, arising from differential success in habitats with different Desmodium and Hylodesmum taxa, is therefore likely to affect Bradyrhizobium diversity patterns at the landscape level. PMID:26130822

  10. 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. PMID:27236566

  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. 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

  13. 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 n

  14. Crystallization and preliminary crystallographic analysis of a haloalkane dehalogenase, DbjA, from Bradyrhizobium japonicum USDA110

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Yukari [Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577 (Japan); Japan Biological Information Research Center (JBIRC), Japan Biological Informatics Consortium (JBIC), 2-42 Aomi, Koto-ku, Tokyo 135-0064 (Japan); Natsume, Ryo [Japan Biological Information Research Center (JBIRC), Japan Biological Informatics Consortium (JBIC), 2-42 Aomi, Koto-ku, Tokyo 135-0064 (Japan); Tsuda, Masataka [Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577 (Japan); Damborsky, Jiri [Loschmidt Laboratories, Masaryk University, Kamenice 5/A4, 625 00 Brno (Czech Republic); Nagata, Yuji [Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577 (Japan); Senda, Toshiya, E-mail: tsenda@jbirc.aist.go.jp [Biological Information Research Center (BIRC), National Institute of Advanced Industrial Science and Technology (AIST), 2-42 Aomi, Koto-ku, Tokyo 135-0064 (Japan); Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577 (Japan)

    2007-04-01

    A haloalkane dehalogenase, DbjA, was crystallized by the hanging-drop vapour-diffusion method using PEG 4000 as a precipitant. The crystal belongs to the orthorhombic system, space group P2{sub 1}2{sub 1}2 and diffracts to 1.75 Å resolution. Haloalkane dehalogenases are key enzymes for the degradation of halogenated aliphatic pollutants. The haloalkane dehalogenase DbjA constitutes a novel substrate-specificity class with high catalytic activity for β-methylated haloalkanes. In order to reveal the mechanism of its substrate specificity, DbjA has been crystallized using the hanging-drop vapour-diffusion method. The best crystals were obtained using the microseeding technique with a reservoir solution consisting of 17–19.5%(w/v) PEG 4000, 0.2 M calcium acetate and 0.1 M Tris–HCl pH 7.7–8.0. The space group of the DbjA crystal is P2{sub 1}2{sub 1}2, with unit-cell parameters a = 212.9, b = 117.8, c = 55.8 Å. The crystal diffracts to 1.75 Å resolution.

  15. Isovaleryl-homoserine lactone, an unusual branched-chain quorum-sensing signal from the soybean symbiont Bradyrhizobium japonicum

    OpenAIRE

    Lindemann, Andrea; Pessi, Gabriella; Schaefer, Amy L.; Mattmann, Margrith E.; Christensen, Quin H.; Kessler, Aline; Hennecke, Hauke; Blackwell, Helen E.; Greenberg, E. Peter; Harwood, Caroline S.

    2011-01-01

    Many species of Proteobacteria communicate by using LuxI-LuxR–type quorum-sensing systems that produce and detect acyl-homoserine lactone (acyl-HSL) signals. Most of the known signals are straight-chain fatty acyl-HSLs, and evidence indicates that LuxI homologs prefer fatty acid-acyl carrier protein (ACP) over fatty acyl-CoA as the acyl substrate for signal synthesis. Two related LuxI homologs, RpaI and BtaI from Rhodopseudomonas palustris and photosynthetic stem-nodulating bradyrhizobia, dir...

  16. Differences in crystallization of two LinB variants from Sphingobium japonicum UT26

    International Nuclear Information System (INIS)

    To study enzyme functionality, two haloalkane dehalogenase variants LinB32 and LinB70 carrying single-point and double-point mutations were constructed and crystallized in different crystallization conditions. Both LinB variants and their complexes with halogenated substrates diffracted to resolutions ranging from 1.6 to 2.8 Å. Haloalkane dehalogenases are microbial enzymes that convert a broad range of halogenated aliphatic compounds to their corresponding alcohols by the hydrolytic mechanism. These enzymes play an important role in the biodegradation of various environmental pollutants. Haloalkane dehalogenase LinB isolated from a soil bacterium Sphingobium japonicum UT26 has a relatively broad substrate specificity and can be applied in bioremediation and biosensing of environmental pollutants. The LinB variants presented here, LinB32 and LinB70, were constructed with the goal of studying the effect of mutations on enzyme functionality. In the case of LinB32 (L117W), the introduced mutation leads to blocking of the main tunnel connecting the deeply buried active site with the surrounding solvent. The other variant, LinB70 (L44I, H107Q), has the second halide-binding site in a position analogous to that in the related haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94. Both LinB variants were successfully crystallized and full data sets were collected for native enzymes as well as their complexes with the substrates 1,2-dibromoethane (LinB32) and 1-bromobutane (LinB70) to resolutions ranging from 1.6 to 2.8 Å. The two mutants crystallize differently from each other, which suggests that the mutations, although deep inside the molecule, can still affect the protein crystallizability

  17. Bradyrhizobium subterraneum sp. nov., a symbiotic nitrogen-fixing bacterium from root nodules of groundnuts.

    Science.gov (United States)

    Grönemeyer, Jann Lasse; Chimwamurombe, Percy; Reinhold-Hurek, Barbara

    2015-10-01

    Seven strains of symbiotic bacteria from root nodules of local races of Bambara groundnut (Vigna subterranea) and peanuts (Arachis hypogaea) grown on subsistence farmers' fields in the Kavango region, Namibia, were previously characterized and identified as a novel group within the genus Bradyrhizobium. To corroborate their taxonomic status, these strains were further characterized using a polyphasic approach. All strains possessed identical 16S rRNA gene sequences with Bradyrhizobium yuanmingense CCBAU 10071T being the most closely related type strain in the 16S rRNA gene phylogenetic analysis, and Bradyrhizobium daqingense CCBAU 15774T in the ITS sequence analysis. Phylogenetic analysis of concatenated glnII-recA-rpoB-dnaK placed the strains in a highly supported lineage distinct from named species of the genus Bradyrhizobium, most closely related to Bradyrhizobium yuanmingense CCBAU 10071T. The species status was validated by results of DNA–DNA hybridization. Phylogenetic analysis of nifH genes placed the novel strains in a group with nifH of ‘Bradyrhizobium arachidis’ CCBAU 051107 that also nodulates peanuts. The combination of phenotypic characteristics from several tests including carbon source utilization and antibiotic resistance could be used to differentiate representative strains from recognized species of the genus Bradyrhizobium. Novel strain 58 2-1T induced effective nodules on V. subterranea, Vigna unguiculata and A. hypogaea, and some strains on Lablab purpureus. Based on the data presented, we conclude that our strains represent a novel species for which the name Bradyrhizobium subterraneum sp. nov. is proposed, with 58 2-1T [ = DSM 100298T = LMG 28792T = NTCCM0016T (Windhoek)] as the type strain. The DNA G+C content of strain 58 2-1T was 64.7 mol% (T m). PMID:26198108

  18. Endophytic Bradyrhizobium spp. isolates from sugarcane obtained through different culture strategies.

    Science.gov (United States)

    Rouws, Luc Felicianus Marie; Leite, Jakson; de Matos, Gustavo Feitosa; Zilli, Jerri Edson; Coelho, Marcia Reed Rodrigues; Xavier, Gustavo Ribeiro; Fischer, Doreen; Hartmann, Anton; Reis, Verônica Massena; Baldani, José Ivo

    2014-08-01

    Brazilian sugarcane has been shown to obtain part of its nitrogen via biological nitrogen fixation (BNF). Recent reports, based on the culture independent sequencing of bacterial nifH complementary DNA (cDNA) from sugarcane tissues, have suggested that members of the Bradyrhizobium genus could play a role in sugarcane-associated BNF. Here we report on the isolation of Bradyrhizobium spp. isolates and a few other species from roots of sugarcane cultivar RB867515 by two cultivation strategies: direct isolation on culture media and capture of Bradyrhizobium spp. using the promiscuous legume Vigna unguiculata as trap-plant. Both strategies permitted the isolation of genetically diverse Bradyrhizobium spp. isolates, as concluded from enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) fingerprinting and 16S ribosomal RNA, nifH and nodC sequence analyses. Several isolates presented nifH phylotypes highly similar to nifH cDNA phylotypes detected in field-grown sugarcane by a culture-independent approach. Four isolates obtained by direct plate cultivation were unable to nodulate V. unguiculata and, based on PCR analysis, lacked a nodC gene homologue. Acetylene reduction assay showed in vitro nitrogenase activity for some Bradyrhizobium spp. isolates, suggesting that these bacteria do not require a nodule environment for BNF. Therefore, this study brings further evidence that Bradyrhizobium spp. may play a role in sugarcane-associated BNF under field conditions. PMID:24992534

  19. Organic Acid Metabolism by Isolated Rhizobium japonicum Bacteroids

    Science.gov (United States)

    Stovall, Iris; Cole, Michael

    1978-01-01

    Rhizobium japonicum bacteroids isolated from soybean (Glycine max L.) nodules oxidized 14C-labeled succinate, pyruvate, and acetate in a manner consistent with operation of the tricarboxylic acid cycle and a partial glyoxylate cycle. Substrate carbon was incorporated into all major cellular components (cell wall + membrane, nucleic acids, and protein). PMID:16660386

  20. 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.

  1. Mosaic origins of Bradyrhizobium legume symbionts on the Caribbean island of Guadeloupe.

    Science.gov (United States)

    Parker, Matthew A; Rousteau, Alain

    2014-08-01

    To analyze geographic affinities of Bradyrhizobium sp. symbionts associated with the diverse legume flora on the Caribbean island of Guadeloupe, 39 isolates from 18 legume genera were compared to a reference set of 269 Bradyrhizobium strains from North America, Central America, Puerto Rico and the Philippines. A multilocus sequence analysis (4192 bp) showed that nucleotide diversity in Guadeloupe equaled or exceeded that found in all other regional Bradyrhizobium populations examined. Bayesian phylogenetic tree analysis grouped the Guadeloupe Bradyrhizobium strains into clades with at least 20 distinct sets of non-Guadeloupe relatives, implying that the island was colonized numerous times from multiple source regions. However, for 18% of the Guadeloupe isolates, inferred geographic affinities for the nifD locus, in the symbiosis island region of the Bradyrhizobium chromosome, conflicted with the source region deduced from a tree based on six concatenated housekeeping genes. Geographic mosaic ancestry was therefore evident among Guadeloupe bradyrhizobia. Horizontal gene transfer subsequent to island colonization appears to have generated strains that carry combinations of genes from disparate source regions. PMID:24747127

  2. Regulation of Polyhydroxybutyrate Synthesis in the Soil Bacterium Bradyrhizobium diazoefficiens.

    Science.gov (United States)

    Quelas, J I; Mesa, S; Mongiardini, E J; Jendrossek, D; Lodeiro, A R

    2016-07-15

    Polyhydroxybutyrate (PHB) is a carbon and energy reserve polymer in various prokaryotic species. We determined that, when grown with mannitol as the sole carbon source, Bradyrhizobium diazoefficiens produces a homopolymer composed only of 3-hydroxybutyrate units (PHB). Conditions of oxygen limitation (such as microoxia, oxic stationary phase, and bacteroids inside legume nodules) were permissive for the synthesis of PHB, which was observed as cytoplasmic granules. To study the regulation of PHB synthesis, we generated mutations in the regulator gene phaR and the phasin genes phaP1 and phaP4 Under permissive conditions, mutation of phaR impaired PHB accumulation, and a phaP1 phaP4 double mutant produced more PHB than the wild type, which was accumulated in a single, large cytoplasmic granule. Moreover, PhaR negatively regulated the expression of phaP1 and phaP4 as well as the expression of phaA1 and phaA2 (encoding a 3-ketoacyl coenzyme A [CoA] thiolases), phaC1 and phaC2 (encoding PHB synthases), and fixK2 (encoding a cyclic AMP receptor protein [CRP]/fumarate and nitrate reductase regulator [FNR]-type transcription factor of genes for microoxic lifestyle). In addition to the depressed PHB cycling, phaR mutants accumulated more extracellular polysaccharides and promoted higher plant shoot dry weight and competitiveness for nodulation than the wild type, in contrast to the phaC1 mutant strain, which is defective in PHB synthesis. These results suggest that phaR not only regulates PHB granule formation by controlling the expression of phasins and biosynthetic enzymes but also acts as a global regulator of excess carbon allocation and symbiosis by controlling fixK2 IMPORTANCE: In this work, we investigated the regulation of polyhydroxybutyrate synthesis in the soybean-nodulating bacterium Bradyrhizobium diazoefficiens and its influence in bacterial free-living and symbiotic lifestyles. We uncovered a new interplay between the synthesis of this carbon reserve polymer

  3. Hypericum japonicum Thunb. ex Murray: Phytochemistry, Pharmacology, Quality Control and Pharmacokinetics of an Important Herbal Medicine

    Directory of Open Access Journals (Sweden)

    Lin-Sheng Liu

    2014-07-01

    Full Text Available Hypericum japonicum Thunb. ex Murray is mainly distributed throughout Asia, Oceania and North America and is used as an important herbal medicine. H. japonicum contains many valuable secondary metabolites, such as flavonoids, phloroglucinols and xanthones and has hepatoprotective, anti-tumor, antibacterial, antiviral, and antioxidant activities and effects on the cardiovascular system and immunity. Coupled with phytochemical and pharmacological research, a series of analytical methods have been developed to evaluate the quality of H. japonicum based on its bioactive components. A pharmacokinetics study involved the absorption of two main flavonoids of H. japonicum in rats. This review aims to present an up-to-date and comprehensive overview of the phytochemistry, pharmacology, quality control and pharmacokinetics of H. japonicum, which should be useful for the greater development of H. japonicum, especially in the development of new drugs and therapeutics for various diseases.

  4. 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; Bromfield, Eden S P

    2014-09-01

    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 18230(T). 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 OO99(T) 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 OO99(T) ( = LMG 26739(T) = HAMBI 3284(T)) as the type strain. The DNA G+C content is 62.6 mol%. PMID:24969302

  5. 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; Bromfield, Eden S P

    2014-09-01

    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 18230(T). 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 OO99(T) 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 OO99(T) ( = LMG 26739(T) = HAMBI 3284(T)) as the type strain. The DNA G+C content is 62.6 mol%.

  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

    2016-01-01

    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.

  7. Schistosoma japonicum-associated morbidity and its mechanisms among children and young adults in the Philippines

    NARCIS (Netherlands)

    Coutinho, H.M.

    2006-01-01

    This thesis presents results from a longitudinal study in Schistosoma japonicum infected children, adolescents and young adults, carried out in the Philippines. The aim of this thesis was to describe nutritional morbidity and hepatic fibrosis in the context of S. japonicum infection and reinfection

  8. Congenital infection of rabbits with Schistosoma japonicum and protective immunity of offspring

    Institute of Scientific and Technical Information of China (English)

    SHI You-en 石佑恩; Abdel-Moneim M Salim 沙里模; NING Chang-xiu 宁长修; GAN Yan 甘燕; ZHU Xiao-hua 朱晓华; PU Ling-yi 卜玲毅

    2004-01-01

    Background Recently congenital infection with Schistosoma japonicum (S. Japonicum) has been domonstrated in pigs, rabbits, mice and dogs. We explored the rabbit as an animal model for the congenital infection of schistosomiasis japonica and assessed the effect of a congenital S. Japonicum infection on the resistance of rabbit kittens to a postnatal challenge infection.Methods Sixteen pregnant New Zealand white rabbits were infected with a single dose of S. Japonicum cercariae. The exposed animals were divided into three groups according to the gestation age at the time of infection. Diagnosis of prenatally acquired S. Japonicum infection in the rabbit kittens was primarily based on serological tests in combination with parasitological and histopathological findings. Congenitally infected kittens were challenged percutaneously with 100 S. Japonicum cercariae to assess the effect of a congenital S. Japonicum infection on kitten resistance to a postnatal challenge infection.Conclusions This study demonstrates the possibility of congenital infection of S. Japonicum in rabbits and the resistance of congenitally infected kittens to a postnatal challenge infection. These results have important implications not only for epidemiological investigations, but also in designing government control programs for schistosomiasis.

  9. Process optimization and kinetic modelling of cyclic (1→3, 1→6)-β-glucans production from Bradyrhizobium japonicum MTCC120.

    Science.gov (United States)

    Nair, Anju V; Gummadi, Sathyanarayana N; Doble, Mukesh

    2016-05-20

    Cyclic (1→3, 1→6)-β-glucans are water soluble, biocompatible polymers with potential applications in food and pharmaceutical industries but have not yet been exploited due to their poor yield. In the present study statistical experimental design methodology was employed to improve their production. Initial screening indicated arabinose and peptone as best carbon and nitrogen source respectively, for glucan production. Arabinose and osmolyte concentrations as well as pH significantly contributed to the glucan production. Central composite design indicated a significant interaction between osmolyte concentration and pH on glucan production. The maximum amount of cyclic glucan produced was 6.7g/L in a 2.5L reactor in batch conditions. The logistic equation for cell growth and Luedeking-Piret equation for glucan production could satisfactorily simulate the batch kinetics data. Cyclic β-glucans could efficiently encapsulate a hydrophobic molecule, curcumin and increase its solubility in water, thus indicating that these glucans have potential as drug delivery systems.

  10. Process optimization and kinetic modelling of cyclic (1→3, 1→6)-β-glucans production from Bradyrhizobium japonicum MTCC120.

    Science.gov (United States)

    Nair, Anju V; Gummadi, Sathyanarayana N; Doble, Mukesh

    2016-05-20

    Cyclic (1→3, 1→6)-β-glucans are water soluble, biocompatible polymers with potential applications in food and pharmaceutical industries but have not yet been exploited due to their poor yield. In the present study statistical experimental design methodology was employed to improve their production. Initial screening indicated arabinose and peptone as best carbon and nitrogen source respectively, for glucan production. Arabinose and osmolyte concentrations as well as pH significantly contributed to the glucan production. Central composite design indicated a significant interaction between osmolyte concentration and pH on glucan production. The maximum amount of cyclic glucan produced was 6.7g/L in a 2.5L reactor in batch conditions. The logistic equation for cell growth and Luedeking-Piret equation for glucan production could satisfactorily simulate the batch kinetics data. Cyclic β-glucans could efficiently encapsulate a hydrophobic molecule, curcumin and increase its solubility in water, thus indicating that these glucans have potential as drug delivery systems. PMID:27046066

  11. Bradyrhizobium kavangense sp. nov., a symbiotic nitrogen-fixing bacterium from root nodules of traditional Namibian pulses.

    Science.gov (United States)

    Lasse Grönemeyer, Jann; Hurek, T; Reinhold-Hurek, Barbara

    2015-12-01

    Eight strains of symbiotic bacteria from root nodules of local races of cowpea (Vigna unguiculata) and Bambara groundnut (Vigna subterranea) grown on subsistence farmers' fields in the Kavango region, Namibia, were previously characterized and identified as a novel group within the genus Bradyrhizobium. To clarify their taxonomic status, these strains were further characterized using a polyphasic approach. In phylogenetic analysis of the 16S rRNA gene sequence the novel group was most closely related to Bradyrhizobium iriomotense EK05T and Bradyrhizobium ingae BR 10250T, and to 'Bradyrhizobium arachidis' CCBAU 051107 in the ITS sequence analysis. Phylogenetic analysis of concatenated glnII-recA-rpoB-dnaK sequences placed the strains in a lineage distinct from named species of the genus Bradyrhizobium. The species status was validated by results of DNA-DNA hybridization. Phylogenetic analysis of nifH and nodC genes placed the novel strains in a group with 'B. arachidis' CCBAU 051107. The combination of phenotypic characteristics from several tests including carbon source utilization and antibiotic resistance could be used to differentiate representative strains from recognized species of the genus Bradyrhizobium. Novel strain 14-3T induces effective nodules on Vigna subterranea, Vigna unguiculata, Arachis hypogaea and Lablab purpureus. Based on the data presented, it is concluded that the strains represent a novel species of the genus Bradyrhizobium, for which the name Bradyrhizobium kavangense sp. nov. is proposed. The type strain is 14-3T [ = DSM 100299T = LMG 28790T = NTCCM 0012T (Windhoek)]. The DNA G+C content of strain 14-3T is 63.8 mol% (Tm). PMID:26446190

  12. Bradyrhizobium vignae sp. nov., a nitrogen-fixing symbiont isolated from effective nodules of Vigna and Arachis.

    Science.gov (United States)

    Grönemeyer, Jann Lasse; Hurek, Thomas; Bünger, Wiebke; Reinhold-Hurek, Barbara

    2016-01-01

    Twenty one strains of symbiotic bacteria from root nodules of local races of cowpea (Vigna unguiculata), Bambara groundnut (Vigna subterranea) and peanuts (Arachis hypogaea) grown on subsistence farmers' fields in the Kavango region of Namibia, were previously characterized as a novel group within the genus Bradyrhizobium. To verify their taxonomic position, the strains were further analysed using a polyphasic approach. 16S rRNA gene sequences were most similar to Bradyrhizobium manausense BR 3351T, with Bradyrhizobium ganzhouense RITF806T being the most closely related type strain in the phylogenetic analysis, and Bradyrhizobium yuanmingense CCBAU 10071T in the ITS sequence analysis. Phylogenetic analysis of concatenated glnII-recA-rpoB-dnaK placed the strains in a highly supported lineage distinct from species of the genus Bradyrhizobium with validly published names; they were most closely related to Bradyrhizobium subterraneum 58 2-1T. The status of the species was validated by results of DNA-DNA hybridization. The combination of phenotypic characteristics from several tests, including carbon source utilization and antibiotic resistance, could be used to differentiate representative strains of species of the genus Bradyrhizobium with validly published names. Novel strain 7-2T induced effective nodules on Vigna subterranea, Vigna unguiculata, Arachis hypogaea and on Lablab purpureus. The DNA G+C content of strain 7-2T was 65.4 mol% (Tm). Based on the data presented, we conclude that these strains represent a novel species for which the name Bradyrhizobium vignae sp. nov. is proposed, with strain 7-2T [LMG 28791T, DSMZ 100297T, NTCCM0018T (Windhoek)] as the type strain. PMID:26463703

  13. Cloning and characterisation of Schistosoma japonicum insulin receptors.

    Directory of Open Access Journals (Sweden)

    Hong You

    Full Text Available BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS: 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

  14. Biodegradation of 5-Nitroanthranilic Acid by Bradyrhizobium sp. Strain JS329▿

    OpenAIRE

    Qu, Yi; Spain, Jim C.

    2010-01-01

    Biodegradation of synthetic compounds has been studied extensively, but the metabolic diversity required for catabolism of many natural compounds has not been addressed. 5-Nitroanthranilic acid (5NAA), produced in soil by Streptomyces scabies, is also the starting material for synthetic dyes and other nitroaromatic compounds. Bradyrhizobium JS329 was isolated from soil by selective enrichment with 5NAA. When grown on 5NAA, the isolate released stoichiometric amounts of nitrite and half of the...

  15. 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

    2016-01-01

    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 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. PMID:27431195

  16. 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.

  17. Isolation and characterization of Bradyrhizobium sp. 224 capable of degrading sulfanilic acid.

    Science.gov (United States)

    Hayase, Nobuki; Fujikawa, Yui; Nakagawa, Katsuhiko; Ushio, Kazutoshi

    2016-08-01

    A bacterial strain (strain 224), which has the ability to utilize sulfanilic acid as a sole source of carbon, was isolated from soil. 16S rRNA gene sequence obtained from strain 224 exhibited 100% identical to that of species in the genus Bradyrhizobium. Strain 224 degraded 4.7 mM of sulfanilic acid and released almost the same molar concentration of sulfate ion.

  18. Tolerância de Bradyrhizobium sp. de mimosoideae à acidez em meio de cultura Tolerance of mimosoideae Bradyrhizobium sp. strains to acidity in culture media

    Directory of Open Access Journals (Sweden)

    Walter Quadros Ribeiro Júnior

    1988-01-01

    Full Text Available Foram realizados testes em meio de cultivo acidificado para avaliar a tolerância de 59 estirpes de Bradyrhizobium sp. isolados de Mimosoideae. As culturas, por via de regra, apresentaram crescimento rápido e alcalinização do meio. Das estirpes testadas, dez apresentaram crescimento em meio com valor de pH 4,6 (três, crescimento rápido; um, médio e seis, lento. Destas, oito não induziram alteração visual na cor do indicador bromotimol-azul incluído no meio. A estirpe SMS-513, uma entre essas oito, promoveu acidificação no meio com valor de pH 6,2, sendo considerada tolerante à acidez. Algumas estirpes cresceram em meio de cultura acidificado, somente com alta concentração inicial de células.Fifty-nine Bradyrhizobium sp. strains isolated from Mimosoideae subfamily of Leguminosae were tested on acidified agar medium. Most strains were found to be fast growing and alcalinized the medium. Ten strains grew on pH 4.6; out of them, three were fast growing, six were slow growing and one was intermediate. Eight of the tested strains did not induce visual changes in the bromothymol-blue indicator. The strain SMS-513 acidified the medium with pH 6.2, and was considered acid tolerant.

  19. [Grey correlation and cluster analysis on relationship between Cercidiphyllum japonicum community and its environment].

    Science.gov (United States)

    Pan, K; Liu, Z

    2001-04-01

    The relationship between 10 years old C. japonicum plantation forest community and its environmental conditions was studied by grey correlation method and cluster analysis. The results showed that there existed closer relations among soil organic matter, available Ca, Fe, Mg and N. Except for N and Mn, all soil available elements tested had no direct impact on the absorption and accumulation of corresponding elements by the community. The tree species in different habitats was found to absorb soil elements selectively to some extent. The factors affecting the basal diameter, DBH, height and biomass of C. japonicum could be lined as soil chemical characteristics > soil physical properties > climate factors, in which, available P had little impact on the growth of C. japonicum. It is suggested that C. japonicum could be restored in the area with higher air moisture and with fertile porous soil between elevation of 2200-2700 m. PMID:11757353

  20. 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.

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

    OpenAIRE

    Yang Jianmei; Fu Zhiqiang; Feng Xingang; Shi Yaojun; Yuan Chunxiu; Liu Jinming; Hong Yang; Li Hao; Lu Ke; Lin Jiaojiao

    2012-01-01

    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...

  2. Erythronium japonicum attenuates histopathological lung abnormalities in a mouse model of ovalbumin-induced asthma.

    Science.gov (United States)

    Seo, Ji-Hye; Bang, Mi-Ae; Kim, Gyeyeop; Cho, Seung Sik; Park, Dae-Hun

    2016-05-01

    Asthma is a chronic lung condition that can induce mucus hypersecretion and pulmonary obstruction and may even cause death, particularly in children and older individuals. Erythronium japonicum (E. japonicum) is a traditional herb used in Korea and East Asian countries that has been found to exert free radical scavenging activity and anti-proliferative effects in human colorectal carcinoma cells. In the present study, we evaluated the anti-asthmatic effects of an extract of E. japonicum in a mouse model of ovalbumin (OVA)‑induced asthma. Female BALB/c mice were sensitized with an intraperitoneal injection of OVA and aluminum hydroxide hydrate on days 1 and 8 and then received the following treatments on days 21 to 25: i) control (no treatment), ii) sterilized tap water (given orally), iii) 1 mg/kg/day dexamethasone (administered orally), iv) 60 mg/kg/day E. japonicum extract, and v) 600 mg/kg/day E. japonicum extract. On the same days, all the mice except those in the control group were challenged 1 h later with nebulized 5% OVA for 30 min. We found that treatment with E. japonicum extract suppressed the OVA-induced increase in the number of white blood cells and decreased the IgE level in the bronchoalveolar lavage fluid samples obtained from the mice. Histopathological analysis of the lung tissues revealed that E. japonicum attenuated the asthma-related morphological changes in the mouse lung tissue, including the increased secretion of mucus in the bronchioles, eosinophil infiltration around the bronchioles and vessels, and goblet cell and epithelial cell hyperplasia. Immunohistochemical analysis revealed that treatment with E. japonicum extract suppressed the OVA-induced proliferation of T helper cells (CD4+) and B cells (CD19+) in the mouse lung tissue. Furthermore, treatment with E. japonicum extract modulated the expression of both T helper 2 cell-related factors [GATA binding protein 3 (GATA-3), tumor necrosis factor

  3. 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

  4. EFECTO DE DIFERENTES MEDIOS DE CULTIVO EN LA MULTIPLICACIÓN CELULAR DE Bradyrhizobium elkanii

    OpenAIRE

    María C. Nápoles; Martínez, J.; Daimy Costales; Gretel Gómez; Ellen Somers

    2006-01-01

    Para crecer, ya sea en la naturaleza o en condiciones de laboratorio, las células necesitan de una fuente de energía, carbono y otros nutrientes, y condiciones tales como la concentración de oxígeno, temperatura y pH. En este trabajo se estudia el comportamiento de dos cepas de Bradyrhizobium elkanii en distintos medios de cultivo. Se evidenciaron diferentes dinámicas de multiplicación celular para cada cepa en los distintos medios, como resultado de su diferenciada composición nutrimental. S...

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

    Directory of Open Access Journals (Sweden)

    Darren J Gray

    Full Text Available BACKGROUND: 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. METHODS AND FINDINGS: 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. CONCLUSIONS: Along with previous epidemiological data, our findings strongly support

  6. The characteristics of NK cells in Schistosoma japonicum-infected mouse spleens.

    Science.gov (United States)

    Li, Lu; Cha, Hefei; Yu, Xiuxue; Xie, Hongyan; Wu, Changyou; Dong, Nuo; Huang, Jun

    2015-12-01

    Natural killer (NK) cells are classic innate immune cells that play roles in many types of infectious disease. Recently, some new characteristics of NK cells were discovered. In this study, C57BL/6 mice were infected with Schistosoma japonicum for 5-6 weeks and lymphocytes were isolated from the spleen to detect some of the NK cell characteristics by multiparametric flow cytometry. The results revealed that the S. japonicum infection induced a large amount of NK cells, although the percentage of NK cells was not increased significantly. At the same time, the results showed that infected mouse splenic NK cells expressed increased levels of CD25 and CD69 and produced more IL-2, IL-4, and IL-17 and less IFN-γ after stimulation with PMA and ionomycin. This meant that NK cells played a role in S. japonicum infection. Moreover, decreased NKG2A/C/E (CD94) expression levels were detected on the surface of NK cells from infected mouse spleens, which might serve as a NK cell activation mechanism. Additionally, high levels of IL-10, but not PD-1, were expressed on the infected mouse NK cells, which implied that functional exhaustion might exist in the splenic NK cells from S. japonicum-infected mice. Collectively, our results suggest that NK cells play important roles in the course of S. japonicum infection. PMID:26319521

  7. Partial protection induced by phage library-selected peptides mimicking epitopes of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Objective To obtain peptide mimicking epitopes of Schistosoma japonicum (S.japonicum) through screening of a phage peptide library and to test their potential for induction of protection. Methods S.japonicum infected sera from Microtus fortis (IMFS) and normal sera from Microtus fortis (NMFS) were used respectively to screen a 12-mers random peptide library by testing the reactivity of anti-S.japonicum serum with the phagotopes. After three rounds of biopanning, the pooled phages were used to immunize mice, after which challenge infection was performed. Results Of 12 randomly picked clones, 10 clones selected using IMFS and 7 clones selected using NMFS were shown to be antigenic. Significant reduction in adult worms (22.6%) and a high reduction (68.9%) in liver eggs were achieved following immunization with phages screened with IMFS. However, no protection was elicited by those selected with NMFS. Conclusion The results show that the phagotopes are both antigenic and immunogenic, suggesting a potential use of phage displayed peptide as novel vaccines against S. japonicum.

  8. Identification and Characterization of Peptides Mimicking the Epitopes of Metalloprotease of Schistosoma Japonicum

    Institute of Scientific and Technical Information of China (English)

    Lianfei Tang; Yuxiao Chen; Linqian Wang; Shunke Zhang; Xianfang Zeng; Xinyuan Yi

    2005-01-01

    In an attempt to isolate and characterize peptides mimicking epitopes of metalloprotease and explore their immunological protection against Schistosoma japonicum (S. japonicum), polyclonal anti-metalloprotease sera was prepared to screen a 12-mer random peptide library to isolate phages binding specially to antisera IgG. Then,phage ELISA, animal immunization, DNA sequencing, Western blotting and enzymatic activity neutralizing analysis were used to characterize the selected phage clones. All of ten randomly picked clones were shown to be positive. Five peptides of different amino acid sequences deduced from DNA sequences were obtained and two of them (peptides 2 and 3) could induce significant reduction (31.0% and 31.8%, respectively) in worm burden and high reduction (52.6% and 54.9%, respectively) in liver eggs per gram (LEPG), while, unexpectedly, others (peptides 1, 4 and 5) could not elicit enough protection against infection of S. japonicum. Peptides 2 and 3 could be recognized by S. japonicum infected mouse sera (IMS) and could elicit neutralizing Abs. The results show that peptides 2 and 3 are antigenic and immunogenic. They are true mimics of epitopes of metalloprotease and useful as novel vaccine candidates against S. japonicum.

  9. Genome Sequence of Bradyrhizobium stylosanthis Strain BR 446T, a Nitrogen-Fixing Symbiont of the Legume Pasture Stylosanthes guianensis.

    Science.gov (United States)

    Delamuta, Jakeline Renata Marçon; Ribeiro, Renan Augusto; Gomes, Douglas Fabiano; Souza, Renata Carolini; Chueire, Ligia Maria Oliveira; Hungria, Mariangela

    2016-01-01

    Bradyrhizobium stylosanthis BR 446(T) is a nitrogen-fixing symbiont of the tropical legume pasture Stylosanthes guianensis Its draft genome contains 8,801,717 bp and 8,239 coding sequences (CDSs). Several putative genes that might confer high competitiveness and saprophytic capacity under the stressful conditions of tropical soils were identified in the genome. PMID:27365354

  10. Functional characterization of a soybean growth stimulator Bradyrhizobium sp. strain SR-6 showing acylhomoserine lactone production.

    Science.gov (United States)

    Ali, Amanat; Ayesha; Hameed, Sohail; Imran, Asma; Iqbal, Mazhar; Iqbal, Javed; Oresnik, Ivan J

    2016-09-01

    A soybean nodule endophytic bacterium Bradyrhizobium sp. strain SR-6 was characterized for production of acyl homoserine lactones (AHLs) as quorum sensing molecules. Mass spectrometry analysis of AHLs revealed the presence of C6-HSL, 3OH-C6-HSL, C8-HSL, C10-HSL, 3oxoC10-HSL, 3oxo-C12-HSL and 3OH-C12-HSL which are significantly different from those reported earlier in soybean symbionts. Purified AHL extracts significantly improved wheat and soybean seedling growth and root hair development along with increased soybean nodulation under axenic conditions. A positive correlation was observed among in vivo nitrogenase and catalase enzyme activities of the strain SR-6. Transmission electron microscopic analysis showed the cytochemical localization of catalase activity within the bacteroids, specifically attached to the peribacteroidal membrane. Root and nodule colonization proved rhizosphere competence of SR-6. The inoculation of SR-6 resulted in increased shoot length (13%), plant dry matter (50%), grain weight (16%), seed yield (20%) and N-uptake (14%) as compared to non-inoculated soybean plants. The symbiotic bacterium SR-6 has potential to improve soybean growth and yield in sub-humid climate of Azad Jammu and Kashmir region of Pakistan. The production and mass spectrometric profiling of AHLs as well as in vivo cytochemical localization of catalase enzyme activity in soybean Bradyrhizobium sp. have never been reported earlier elsewhere before our these investigations.

  11. Functional characterization of a soybean growth stimulator Bradyrhizobium sp. strain SR-6 showing acylhomoserine lactone production.

    Science.gov (United States)

    Ali, Amanat; Ayesha; Hameed, Sohail; Imran, Asma; Iqbal, Mazhar; Iqbal, Javed; Oresnik, Ivan J

    2016-09-01

    A soybean nodule endophytic bacterium Bradyrhizobium sp. strain SR-6 was characterized for production of acyl homoserine lactones (AHLs) as quorum sensing molecules. Mass spectrometry analysis of AHLs revealed the presence of C6-HSL, 3OH-C6-HSL, C8-HSL, C10-HSL, 3oxoC10-HSL, 3oxo-C12-HSL and 3OH-C12-HSL which are significantly different from those reported earlier in soybean symbionts. Purified AHL extracts significantly improved wheat and soybean seedling growth and root hair development along with increased soybean nodulation under axenic conditions. A positive correlation was observed among in vivo nitrogenase and catalase enzyme activities of the strain SR-6. Transmission electron microscopic analysis showed the cytochemical localization of catalase activity within the bacteroids, specifically attached to the peribacteroidal membrane. Root and nodule colonization proved rhizosphere competence of SR-6. The inoculation of SR-6 resulted in increased shoot length (13%), plant dry matter (50%), grain weight (16%), seed yield (20%) and N-uptake (14%) as compared to non-inoculated soybean plants. The symbiotic bacterium SR-6 has potential to improve soybean growth and yield in sub-humid climate of Azad Jammu and Kashmir region of Pakistan. The production and mass spectrometric profiling of AHLs as well as in vivo cytochemical localization of catalase enzyme activity in soybean Bradyrhizobium sp. have never been reported earlier elsewhere before our these investigations. PMID:27242370

  12. Swimming performance of Bradyrhizobium diazoefficiens is an emergent property of its two flagellar systems

    Science.gov (United States)

    Quelas, J. Ignacio; Althabegoiti, M. Julia; Jimenez-Sanchez, Celia; Melgarejo, Augusto A.; Marconi, Verónica I.; Mongiardini, Elías J.; Trejo, Sebastián A.; Mengucci, Florencia; Ortega-Calvo, José-Julio; Lodeiro, Aníbal R.

    2016-01-01

    Many bacterial species use flagella for self-propulsion in aqueous media. In the soil, which is a complex and structured environment, water is found in microscopic channels where viscosity and water potential depend on the composition of the soil solution and the degree of soil water saturation. Therefore, the motility of soil bacteria might have special requirements. An important soil bacterial genus is Bradyrhizobium, with species that possess one flagellar system and others with two different flagellar systems. Among the latter is B. diazoefficiens, which may express its subpolar and lateral flagella simultaneously in liquid medium, although its swimming behaviour was not described yet. These two flagellar systems were observed here as functionally integrated in a swimming performance that emerged as an epistatic interaction between those appendages. In addition, each flagellum seemed engaged in a particular task that might be required for swimming oriented toward chemoattractants near the soil inner surfaces at viscosities that may occur after the loss of soil gravitational water. Because the possession of two flagellar systems is not general in Bradyrhizobium or in related genera that coexist in the same environment, there may be an adaptive tradeoff between energetic costs and ecological benefits among these different species. PMID:27053439

  13. Chemical composition and antimicrobial activity of essential oil isolated from the cultured mycelia of Ganoderma japonicum

    Institute of Scientific and Technical Information of China (English)

    Dandan Liu; Zheng Hu; Zhigang Liu; Bo Yang; Wenjuan Tu; Liang Li

    2009-01-01

    Objective:To explore a new natural antibiotic. Methods:The chemical composition of the essential oil from Ganoderma japonicum (G. japonicum) mycelia was analyzed by gas chromatography-mass spectrometry(GC-MS). The antimicrobial activity of the oil was evaluated against eighteen microorganisms, including bacteria, mildew and yeast by using a disc diffusion method. Furthermore, the minimum inhibitory concentrations(MIC) and the minimum bactericidal concentrations(MBC) of the essential oil against twelve clinical pathogens were determined. Results:The main components of the oil were nerolidol, decadienal, linaiool and benzyl alcohol. The antimicrobial results indicated that the oil inhibited all the tested bacterium, especially Methicillin-resistant Staphylococcus aureus (MRSA) in which the antibacterial activity exhibited a MBC of 1.03 mg/ml. Conclusion: The essential oil of G.japonicum mycelium has significant inhibitory activity. It is a potential medicinal resource that can be used as a natural antibiotic.

  14. Morphogenesis of Pistillate Flowers of Cercidiphyllum japonicum (Cercidiphyllaceae)

    Institute of Scientific and Technical Information of China (English)

    Xiao-Ling Yan; Yi Ren; Xian-Hua Tian; Xiao-Hui Zhang

    2007-01-01

    Floral morphogenesis and the development of Cercidiphyllumjaponicum Sieb. et Zucc. were observed by scanning electron microscopy (SEM). The results showed that the pistillate inflorescences were congested spikes with the flowers arranged opposite. Great differences between the so-called "bract" and the vegetative leaf were observed both in morphogenesis and morphology. In morphogenesis, the "bract" primordium is crescent-shaped, truncated at the apex and not conduplicate,has no stipule primordium at the base but does have some inconspicuous teeth in the margin that are not glandular. The leaf primordium is triangular, cycloidal at the apex, conduplicate, has two stipule primordia at the base, has one gland-tooth at the apex occurring at first and some gland-teeth in the margin that occur later. In morphology, the "bract" is also different to the vegetative leaf in some characteristics that were also illustrated in the present paper. Based on the hypothesis that the bract is more similar to the vegetative leaf than the tepal, we considered that the so-called "bract" of C. japonicum might be the tepal of the pistillate flower in morphological nature. Therefore, each pistillate flower contains a tepal and a carpel.We did not find any trace of other floral organs in the morphogenesis of the pistillate flower. Therefore we considered that the unicarpellate status of extant Cercidiphyllum might be to highly reduce and advance characteristics that make the extant Cercidiphyllum isolated from both fossil Cercidiphyllum-like plants and its extant affinities.

  15. 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.

  16. 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.

  17. Constructing the recombinant plasmid-pLXSN-SjYF-of Yolk Ferritin gene of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    ZhouJunmei; YuXinbing; WuZhongdao; ZhengYinan; LiYan

    1999-01-01

    TO construct the recotabinant plasmid —— pLXSN-SjYF to prepare for expression and DNA vaccine of Schistosoma japonicum gene. Methods :Mnpliffing DNA fragment coding yolk fereltin from a female adult Sehistosoma japonicum DNAs by PCR. The fragment was inseted into pLXSN retrovirus vector by digesting with restrictive enzymes and linking reactions. The positive clone was screened on LB plates contmaining amplcillin asld identified by restrictive enzymes digestion and PCR amplification, Resttlts The specific DNA fragmem SjYF was amplified from the female adult SjDNAs. IaLXSN-SjYF was constructed successfully and the further research will be carried out.

  18. Intercropping Sorghum (Sorghum bicolor L.) and Cowpea (Vigna unguiculata L.): Effect of Bradyrhizobium Inoculation and Fertilization on Minerals Composition of Sorghum Seeds

    OpenAIRE

    Ekhlas M. Musa; Elsiddig A. E. Elsheikh; Isam A. Mohamed Ahmed; Babiker, Elfadil E.

    2012-01-01

    This investigation was carried out in order to gain a better understanding of the effect of Bradyrhizobium inoculation and fertilization on the nutritional values of macro- and micronutrients of sorghum intercropped with cowpea in the field under rainfed conditions. The results of the present study demonstrated that intercropping, Bradyrhizobium inoculation, and N and P fertilization had significant effect on the mineral status of sorghum seeds. Intercropping significantly ( ≤ 0 . 0 5 ) inc...

  19. 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.

  20. EFECTO DE DIFERENTES MEDIOS DE CULTIVO EN LA MULTIPLICACIÓN CELULAR DE Bradyrhizobium elkanii

    Directory of Open Access Journals (Sweden)

    María C. Nápoles

    2006-01-01

    Full Text Available Para crecer, ya sea en la naturaleza o en condiciones de laboratorio, las células necesitan de una fuente de energía, carbono y otros nutrientes, y condiciones tales como la concentración de oxígeno, temperatura y pH. En este trabajo se estudia el comportamiento de dos cepas de Bradyrhizobium elkanii en distintos medios de cultivo. Se evidenciaron diferentes dinámicas de multiplicación celular para cada cepa en los distintos medios, como resultado de su diferenciada composición nutrimental. Se destacó para las dos cepas estudiadas el medio Propagación modificado.

  1. Effect of Bradyrhizobium sp.Inoculation on Biomass and Nodulation of Cowpea

    Institute of Scientific and Technical Information of China (English)

    ZHAOYUHUA; C.F.FONJI; 等

    1997-01-01

    Vigna unguiculata(L.)Walp (cowpea)is a tropical legume of very high nutrive and economic values.A laboratory experiment.was conducted to evaluate the BNF (biological nitrogen fixation) effciency of 4 strains of Bradyrhizobium sp.(VUXYI,VUIE1,VUMDS1 and VUID1).From the results obtained these strains were classified into efficiency groups,with VUID1 being the most efficient and VUXY1 the least efficient.the effect of inoculation was tested on 2 cowpea varieties in a field experiment.Remarkable increases in nodulation biomass and crop yield were observed.An increase from 58% ti 81% in dry seed was obtained for the two varieties.A significance test revealed a signifecantly positive correlation between nodulation and biomass.

  2. Screening and identification of DNA aptamers toward Schistosoma japonicum eggs via SELEX

    Science.gov (United States)

    Long, Yuqian; Qin, Zhiqiang; Duan, Minlan; Li, Shizhu; Wu, Xiaoqiu; Lin, Wei; Li, Jianglin; Zhao, Zilong; Liu, Jing; Xiong, Dehui; Huang, Yi; Hu, Xiaoxiao; Yang, Chao; Ye, Mao; Tan, Weihong

    2016-01-01

    Schistosomiasis is a major parasitic disease caused by blood flukes of the genus Schistosoma. Several million people all over the world are estimated to suffer from severe morbidity as a consequence of schistosomiasis. The worm’s eggs, which cause the symptoms of schistosomiasis, are generally used to diagnose the disease. In this study, we employed egg-based systematic evolution of ligands by exponential enrichment (egg-SELEX) and identified a panel of ssDNA aptamers specifically binding to eggs derived from S. japonicum. Among these, two aptamers LC6 and LC15 exhibited strong binding to and specific recognition of S. japonicum eggs, but not eggs from Fasciolopsis buski, Enterobius, Ascaris or Clonorchis sinensis. Furthermore, tissue imaging results revealed that LC15 could recognize S. japonicum eggs laid in liver tissues with a detection ratio of 80.5%. Collectively, therefore, we obtained useful aptamers specifically recognizing S. japonicum eggs, which will facilitate the development of an effective tool for both schistosomiasis diagnosis and drug delivery. PMID:27121794

  3. Isolation of genes (nif/hup cosmids) involved in hydrogenase and nitrogenase activities in Rhizobium japonicum.

    Science.gov (United States)

    Hom, S S; Graham, L A; Maier, R J

    1985-03-01

    Recombinant cosmids containing a Rhizobium japonicum gene involved in both hydrogenase (Hup) and nitrogenase (Nif) activities were isolated. An R. japonicum gene bank utilizing broad-host-range cosmid pLAFR1 was conjugated into Hup- Nif- R. japonicum strain SR139. Transconjugants containing the nif/hup cosmid were identified by their resistance to tetracycline (Tcr) and ability to grow chemoautotrophically (Aut+) with hydrogen. All Tcr Aut+ transconjugants possessed high levels of H2 uptake activity, as determined amperometrically. Moreover, all Hup+ transconjugants tested possessed the ability to reduce acetylene (Nif+) in soybean nodules. Cosmid DNAs from 19 Hup+ transconjugants were transferred to Escherichia coli by transformation. When the cosmids were restricted with EcoRI, 15 of the 19 cosmids had a restriction pattern with 13.2-, 4.0-, 3.0-, and 2.5-kilobase DNA fragments. Six E. coli transformants containing the nif/hup cosmids were conjugated with strain SR139. All strain SR139 transconjugants were Hup+ Nif+. Moreover, one nif/hup cosmid was transferred to 15 other R. japonicum Hup- mutants. Hup+ transconjugants of six of the Hup- mutants appeared at a frequency of 1.0, whereas the transconjugants of the other nine mutants remained Hup-. These results indicate that the nif/hup gene cosmids contain a gene involved in both nitrogenase and hydrogenase activities and at least one and perhaps other hup genes which are exclusively involved in H2 uptake activity.

  4. 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...

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

    Directory of Open Access Journals (Sweden)

    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.

  6. De novo transcriptome assembly of a fern, Lygodium japonicum, and a web resource database, Ljtrans DB.

    Science.gov (United States)

    Aya, Koichiro; Kobayashi, Masaaki; Tanaka, Junmu; Ohyanagi, Hajime; Suzuki, Takayuki; Yano, Kenji; Takano, Tomoyuki; Yano, Kentaro; Matsuoka, Makoto

    2015-01-01

    During plant evolution, ferns originally evolved as a major vascular plant with a distinctive life cycle in which the haploid and diploid generations are completely separated. However, the low level of genetic resources has limited studies of their physiological events, as well as hindering research on the evolutionary history of land plants. In this study, to identify a comprehensive catalog of transcripts and characterize their expression traits in the fern Lygodium japonicum, nine different RNA samples isolated from prothalli, trophophylls, rhizomes and sporophylls were sequenced using Roche 454 GS-FLX and Illumina HiSeq sequencers. The hybrid assembly of the high-quality 454 GS-FLX and Illumina HiSeq reads generated a set of 37,830 isoforms with an average length of 1,444 bp. Using four open reading frame (ORF) predictors, 38,142 representative ORFs were identified from a total of 37,830 transcript isoforms and 95 contigs, which were annotated by searching against several public databases. Furthermore, an orthoMCL analysis using the protein sequences of L. japonicum and five model plants revealed various sets of lineage-specific genes, including those detected among land plant lineages and those detected in only L. japonicum. We have also examined the expression patterns of all contigs/isoforms, along with the life cycle of L. japonicum, and identified the tissue-specific transcripts using statistical expression analyses. Finally, we developed a public web resource, the L. japonicum transcriptome database at http://bioinf.mind.meiji.ac.jp/kanikusa/, which provides important opportunities to accelerate molecular research in ferns.

  7. Immunization of mice with cells from juvenile worms of Schistosoma japonicum provides immunoprotection against schistosomiasis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    To validate the protective efficacy against schistosomiasis by immunization with cells from juvenile Schistosoma japonicum in a murine model and to analyze possible factors related to protection, in this study, two independent repeated vaccination trials were performed. After three subcutaneous vaccinations, in trial one, in the absence of adjuvant, primary juvenile worm cells (pJCs) from S. japonicum induced remarkable average reductions in worm burden (54.3%), liver eggs per gram (LEPG) load (59.8%) as well as egg granulomas size (66.5%) compared to PBS control group (P<0.01), which were significantly higher than those elicited by fractions of juvenile worm cells (JCFs) or fractions of juvenile worms (JWFs) (P<0.05). Non-cell components of worms (WNCs) showed no significant protection. In trial two, compared to PBS control group, significant protective effect was also observed for cultured juvenile worm cells (cJCs) from S. japonicum with 58.4% worm reduction and 68.1% LEPG reduction (P<0.01). However, cultured adult worms cells (cACs) showed significantly higher worm burden (P<0.05) and egg burden (P<0.01) when compared to cJCs. Immunological analysis of trial two revealed that cJCs engendered a Th1-biased mixed Th1/Th2 type of immune response while cACs elicited a Th2-type response. Our data indicated that immunization with both primary and cultured cells from S. japonicum juvenile worms provided high immunoprotection, for which the physical character of immunogens, stage-specific parasite and the type of immune response induced might be responsible, suggesting that vaccination with whole cells from S. japonicum larvae is a promising approach to produce protective immunity against schistosomiasis.

  8. Respuesta de diferentes variedades de cereales a la inoculación con Bradyrhizobium sp. Response of different varieties of cereals to inoculation with Bradyrhizobium sp.

    Directory of Open Access Journals (Sweden)

    Carlos José Bécquer Granados

    2012-02-01

    Full Text Available Se llevaron a cabo dos experimentos de campo durante 2009, en la Estación Experimental de Pastos y Forrajes Sancti Spiritus, Cuba (2 1º 53' 00" latitud norte, 79° 21' 25" longitud oeste y 40 msnm, para determinar la respuesta de diferentes variedades de cereales a la biofertilización con Bradyrhizobium sp. La preparación de los inóculos y la inoculación de las semillas, se efectuaron según metodologías prestablecidas para este campo de estudio. El diseño experimental fue a través de parcelas divididas y 4 réplicas por tratamiento. Al control fertilizado se le aplicó nitrógeno (150 kg N ha-¹ y se evaluaron diferentes variables agronómicas. Se aplicó análisis de varianza bifactorial; las diferencias entre medias se determinaron por la dócima de comparación de Duncan y t-student. En el experimento con Triticum se concluye que los tratamientos con mejores respuestas, fueron Triticale-inoculado y a IRM37-inoculado y que Triticum secale superó en peso seco raíz y en longitud del tallo a T. aestivum, lo que hace a esta primera especie promisoria para condiciones de sequía. En el experimento con Zea mays se concluye que aunque en peso seco aéreo los mejores tratamientos correspondieron a la fertilización química, en peso seco de la mazorca el tratamiento Canilla-inoculado, presentó valores estadísticamente iguales al de los tratamientos fertilizados, por lo que se considera positiva la respuesta de esta variedad a la biofertilización. No obstante, no se descarta la influencia de las bacterias rizosféricas autóctonas en algunas de las variables estudiadas.Two field experiments were performed in 2009 at the Experimental Station of Pastures and Forages Sancti Spiritus, Cuba (21° 53' 00" north, 79° 21' 25" west longitude and 40 m to determine the response of different varieties of cereals to bio-fertilization with Bradyrhizobium sp. The preparations of inocula and seed inoculation were made according to preset methodologies

  9. Ultrastructural Observation and Gene Expression Profiling of Schistosoma japonicum Derived from Two Natural Reservoir Hosts, Water Buffalo and Yellow Cattle

    OpenAIRE

    Jianmei Yang; Xingang Feng; Zhiqiang Fu; Chunxiu Yuan; Yang Hong; Yaojun Shi; Min Zhang; Jinming Liu; Hao Li; Ke Lu; Jiaojiao Lin

    2012-01-01

    Water buffalo and yellow cattle are the two of the most important natural reservoir hosts for Schistosoma japonicum in endemic areas of China, although their susceptibility differs, with water buffalo being less conducive to the growth and development of S. japonicum. Results from the current study show that the general morphology and ultrastructure of adult schistosomes derived from the two hosts also differed. Using high-throughput microarray technology, we also compared the gene expression...

  10. Effect of ozonide OZ418 against Schistosoma japonicum harbored in mice.

    Science.gov (United States)

    Xue, Jian; Wang, Xiaofang; Dong, Yuxiang; Vennerstrom, Jonathan L; Xiao, Shu-hua

    2014-09-01

    The in vitro and in vivo efficacies of ozonide carboxylic acid OZ418 against Schistosoma japonicum were investigated. For in vitro experiments, juvenile (14-day-old) and adult schistosomes were collected from mice infected with 80-100 S. japonicum cercariae for 14 and 35 days post-infection and the worms were maintained in Roswell Park Memorial Institute (RPMI) 1640 supplemented by 10% calf serum. Against 35-day-old adult S. japonicum, OZ418 resulted in weakened worm motor activity, injury to the worm body, emergence of vacuoles along the worm surface, and death. A similar outcome was seen in 14-day-old juvenile S. japonicum exposed to OZ418. Ineffective concentrations (1, 5, and 10 μg/mL) of OZ418 also interacted with hemin to significantly increase the killing effect against adult schistosomes. The LC50 value of OZ418 against juvenile (14-day-old) and adult schistosomes were identical--16.2 μg/mL, whereas the corresponding LC95 values were 30.7 and 22.7 μg/mL, respectively. Treatment of adult and juvenile (14-day-old) S. japonicum-infected mice with single 200-400-mg/kg oral doses of OZ418 produced total worm burden reductions of 68.5-84.1 and 37.5-50.9%, respectively. Further study showed that in mice infected with various stages of schistosomes and treated with a single oral OZ418 400 mg/kg, poor efficacy was seen in the 3-h-old juvenile worm group, while 14-day-old and 21-day-old juvenile worm groups exhibited less efficacy with total worm burden reductions of 42.6-52.4%. On the other hand, similar and higher total worm burden reductions (64.2-76.0%) were seen in the 7-day-old juvenile worm group and 28-day-old as well as 35-day-old adult worm groups. Furthermore, the mean worm burden reductions of the 7-day-old juvenile worm group and 35-day-old adult worm group were statistically significantly higher than that of the 14-day-old or 21-day-old juvenile worm group (P < 0.01 or <0.05). These data suggest that OZ418 has promising efficacy against 7-day

  11. 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.

  12. Antischistosomal activity of N,N'-arylurea analogs against Schistosoma japonicum.

    Science.gov (United States)

    Yao, Houzong; Liu, Fengyou; Chen, Jinglei; Li, Yan; Cui, Jinhao; Qiao, Chunhua

    2016-03-01

    Although the antischistosomal activities of N,N'-arylurea analogs were reported, systematic structure-activity relationships have not been conducted. In this Letter, we reported the design, synthesis and evaluation of 45 N,N'-arylurea analogs. Among these prepared compounds, 13 compounds were urea linker modified and 32 were N,N'-arylurea derivatives. The activity evaluation revealed 12 analogs exhibited IC50 values lower than 22.6μM, and 7 of them had IC50 less than 10μM against the juvenile Schistosoma japonicum in vitro. Their worm killing potency was even higher against adult worm. Unfortunately, low to moderate worm burden reduction of 0-33.4% was recorded after administration of a single oral dose of 200mg/kg or 400mg/kg to mice harboring S. japonicum.

  13. The Schistosoma japonicum genome reveals features of host-parasite interplay

    OpenAIRE

    Zhou, Yan; Zheng, Huajun; Chen, Xiangyi; Zhang, Lei; WANG Kai; Guo, Jing; Huang, Zhen; Zhang, Bo; Huang, Wei; Jin, Ke; Tonghai, Dou; Hasegawa, Masami; Wang, Li; Zhang, Yuan; Zhou, Jie

    2009-01-01

    Schistosoma japonicum is a parasitic flatworm that causes human schistosomiasis, a significant cause of morbidity in China and the Philippines. Here we present a draft genomic sequence for the worm, which is the first reported for any flatworm, indeed for the superphylum Lophotrochozoa. The genome provides a global insight into the molecular architecture and host interaction of this complex metazoan pathogen, revealing that it can exploit host nutrients, neuroendocrine hormones and signaling ...

  14. Transmissible Resistance to Penicillin G, Neomycin, and Chloramphenicol in Rhizobium japonicum1

    Science.gov (United States)

    Cole, Michael A.; Elkan, Gerald H.

    1973-01-01

    The genetic basis for resistance to a number of antibiotics was examined in Rhizobium japonicum. Resistance to penicillin G, neomycin, and chloramphenicol appears to be mediated by an extrachromosomal element similar to that found in the Enterobacteriaceae. Resistance to these antibiotics was eliminated from cells by treatment with acridine orange, and resistance to all three antibiotics could be transferred en bloc to Agrobacterium tumefaciens under conditions excluding transformation or transduction as possible genetic mechanisms. PMID:4491197

  15. Flavonol tetraglycosides and other constituents from leaves of Styphnolobium japonicum (Leguminosae) and related taxa.

    Science.gov (United States)

    Kite, Geoffrey C; Stoneham, Charlotte A; Veitch, Nigel C

    2007-05-01

    Two flavonol tetraglycosides comprising a trisaccharide at C-3 and a monosaccharide at C-7 were isolated from the leaves of Styphnolobium japonicum (L.) Schott and characterised as the 3-O-alpha-rhamnopyranosyl(1-->2)[alpha-rhamnopyranosyl(1-->6)]-beta-glucopyranoside-7-O-alpha-rhamnopyranosides of quercetin and kaempferol. The 3-O-alpha-rhamnopyranosyl(1-->2)[alpha-rhamnopyranosyl(1-->6)]-beta-galactopyranoside-7-O-alpha-rhamnopyranoside of kaempferol, the 3-O-alpha-rhamnopyranosyl(1-->2)[alpha-rhamnopyranosyl(1-->6)]-beta-glucopyranosides of kaempferol and quercetin and the 3-O-alpha-rhamnopyranosyl(1-->2)[alpha-rhamnopyranosyl(1-->6)]-beta-galactopyranoside of kaempferol were also obtained from this species for the first time. Some or all of these flavonol tetra- and triglycosides were detected in 17 of 18 specimens of S. japonicum examined from living and herbarium material, although the most abundant flavonoid in the leaves was generally quercetin 3-O-alpha-rhamnopyranosyl(1-->6)-beta-glucopyranoside (rutin). The triglycosides, but not the tetraglycosides, were detected in herbarium specimens of Styphnolobium burseroides M. Sousa, Rudd & Medrano and Styphnolobium monteviridis M. Sousa & Rudd, but specimens of Styphnolobium affine (Torrey & A. Gray) Walp. contained a different profile of flavonol glycosides. The flavonol tetra- and triglycosides of S. japonicum were also present in leaves of Cladrastis kentukea (Dum. Cours.) Rudd, a representative of a genus placed close to Styphnolobium in current molecular phylogenies. An additional constituent obtained from leaves of Styphnolobium japonicum was identified as the maltol derivative, 3-hydroxy-2-methyl-4H-pyran-4-one 3-O-(4'-O-p-coumaroyl-6'-O-(3-hydroxy-3-methylglutaroyl))-beta-glucopyranoside. PMID:17462679

  16. Diversification of Schistosoma japonicum in Mainland China revealed by mitochondrial DNA.

    Directory of Open Access Journals (Sweden)

    Qin Ping Zhao

    Full Text Available BACKGROUND: Schistosoma japonicum still causes severe parasitic disease in mainland China, but mainly in areas along the Yangtze River. However, the genetic diversity in populations of S. japonicum has not been well understood across its geographical distribution, and such data may provide insights into the epidemiology and possible control strategies for schistosomiasis. METHODOLOGY/PRINCIPAL FINDINGS: In this study infected Oncomelania snails were collected from areas in the middle and lower (ML reaches of the Yangtze River, including Hubei, Hunan, Anhui, Jiangxi and Jiangsu provinces, and in the upper reaches of the river, including Sichuan and Yunnan provinces in southwest (SW China. The adult parasites obtained from experimentally infected mice using isolated cercariae were sequenced individually for several fragments of mitochondrial regions, including Cytb-ND4L-ND4, 16S-12S and ND1. Populations in the ML reaches exhibited a relatively high level of diversity in nucleotides and haplotypes, whereas a low level was observed for populations in the SW, using either each single fragment or the combined sequence of the three fragments. Pairwise analyses of F-statistics (Fst revealed a significant genetic difference between populations in the ML reaches and those in the SW, with limited gene flow and no shared haplotypes in between. It is rather obvious that genetic diversity in the populations of S. japonicum was significantly correlated with the geographical distance, and the geographical separation/isolation was considered to be the major factor accounting for the observed difference between populations in the ML reaches and those in the SW in China. CONCLUSIONS: S. japonicum in mainland China exhibits a high degree of genetic diversity, with a similar pattern of genetic diversity as observed in the intermediate host snails in the same region in China.

  17. Transcriptome Bioinformatical Analysis of Vertebrate Stages of Schistosoma japonicum Reveals Alternative Splicing Events

    OpenAIRE

    Xinye Wang; Xindong Xu; Xingyu Lu; Yuanbin Zhang; Weiqing Pan

    2015-01-01

    Alternative splicing is a molecular process that contributes greatly to the diversification of proteome and to gene functions. Understanding the mechanisms of stage-specific alternative splicing can provide a better understanding of the development of eukaryotes and the functions of different genes. Schistosoma japonicum is an infectious blood-dwelling trematode with a complex lifecycle that causes the tropical disease schistosomiasis. In this study, we analyzed the transcriptome of Schistoso...

  18. Exosome-like vesicles derived by Schistosoma japonicum adult worms mediates M1 type immune- activity of macrophage.

    Science.gov (United States)

    Wang, Lifu; Li, Zhitao; Shen, Jia; Liu, Zhen; Liang, Jinyi; Wu, Xiaoying; Sun, Xi; Wu, Zhongdao

    2015-05-01

    Exosomes are 30-100-nm membrane vesicles of endocytic origin that are released into the extracellular space upon fusion of the multi-vesicular bodies (MVB) with the plasma membrane, while initial studies described that the role of exosomes was a reticulocyte cargo-disposal mechanism allowing remodeling of the plasma membrane during the maturation of reticulocytes to erythrocytes. Recent studies indicate that exosomes are secreted by most cells and pathogens and play an important role in intercellular signaling and exert regulatory function by carrying bioactive molecules. As numerous pathogens, adult worm of Schistosoma japonicum (S. japonicum) reside in mesenteric veins of definitive host including man and mammal animals. It was reported that the worms or the eggs also have specialized secretion systems to export effector proteins or other molecules into host target cells. However, the mechanisms involved remained unclear. This study investigated the isolation of the exosome-like vesicles secreted by S. japonicum adult worms and its immune activity on microphage in vitro. In this report, we identified exosome-based secretion as a new mechanism for protein secretion by S. japonicum. Electron microscopy tomography revealed the previously unidentified ultrastructural detail of exosome-like vesicles with high resolution; they were found to be typical spherical shape and to have a diverse population that varies in size of 30-100 nm. Exosome-like vesicles isolated from S. japonicum contained a significantly different protein compared with debris pelleted and the apoptosis body. We also demonstrate that macrophages were preferentially differentiated into the M1 subtype while being treated with S. japonicum exosome-like vesicles. This study reveals there are exosome-like vesicles derived by S. japonicum adult worms, and the exosome-like vesicles can mediate M1-type immune- activity of macrophage.

  19. 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.

  20. 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.

  1. Functional diversity of Cercidiphyllum japonicum, communities in the Shennongjia Reserve, central China

    Institute of Scientific and Technical Information of China (English)

    Jintun Zhang; Bin Zhang; Zhiying Qian

    2015-01-01

    Functional diversity is significant to ecological processes of plant communities. We analyzed the variation of functional diversity of endangered species, Cercidi-phyllum japonicum, communities along an elevational gradient in the Shennongjia Reserve, central China. Sixty plots of 10 × 20 m from 1,350 to 2,050 m were set up and species composition, traits and environmental variables were measured and recorded. These data were analyzed using five functional diversity indices, functional attribute diversity, modified functional attribute diversity, plot based functional diversity, community based functional diversity and Rao’s functional diversity indices (Rao’s index), Functional diversities of C. japonicum communities were rich and varied greatly. Functional diversity declined non-linearly with increasing elevation. Functional diversity was significantly correlated with species richness and hetero-geneity. Elevation was a key environmental variable influencing functional diversity and species diversity. The five functional diversity indices were all effective for measuring functional diversity of communities. Functional diversity can be used as an indicator of conservation effi-ciency of endangered species such as C. japonicum.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Indian Academy of Sciences (India)

    Marí­a Soledad Figueredo; María Laura Tonellie; Tania Taurian; Jorge Angelini; Fernando Ibañez; Lucio Valetti; Vanina Muñoz; Marí­a Soledad Anzuay; Liliana Ludueña; Adriana Fabra

    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.

  4. A Peptidoglycan-Remodeling Enzyme Is Critical for Bacteroid Differentiation in Bradyrhizobium spp. During Legume Symbiosis.

    Science.gov (United States)

    Gully, Djamel; Gargani, Daniel; Bonaldi, Katia; Grangeteau, Cédric; Chaintreuil, Clémence; Fardoux, Joël; Nguyen, Phuong; Marchetti, Roberta; Nouwen, Nico; Molinaro, Antonio; Mergaert, Peter; Giraud, Eric

    2016-06-01

    In response to the presence of compatible rhizobium bacteria, legumes form symbiotic organs called nodules on their roots. These nodules house nitrogen-fixing bacteroids that are a differentiated form of the rhizobium bacteria. In some legumes, the bacteroid differentiation comprises a dramatic cell enlargement, polyploidization, and other morphological changes. Here, we demonstrate that a peptidoglycan-modifying enzyme in Bradyrhizobium strains, a DD-carboxypeptidase that contains a peptidoglycan-binding SPOR domain, is essential for normal bacteroid differentiation in Aeschynomene species. The corresponding mutants formed bacteroids that are malformed and hypertrophied. However, in soybean, a plant that does not induce morphological differentiation of its symbiont, the mutation does not affect the bacteroids. Remarkably, the mutation also leads to necrosis in a large fraction of the Aeschynomene nodules, indicating that a normally formed peptidoglycan layer is essential for avoiding the induction of plant immune responses by the invading bacteria. In addition to exopolysaccharides, capsular polysaccharides, and lipopolysaccharides, whose role during symbiosis is well defined, our work demonstrates an essential role in symbiosis for yet another rhizobial envelope component, the peptidoglycan layer. PMID:26959836

  5. 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

  6. 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.

  7. 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

    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......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......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...

  8. Complete Genome Sequence of Bradyrhizobium sp. Strain CCGE-LA001, Isolated from Field Nodules of the Enigmatic Wild Bean Phaseolus microcarpus.

    Science.gov (United States)

    Servín-Garcidueñas, Luis E; Rogel, Marco A; Ormeño-Orrillo, Ernesto; Zayas-Del Moral, Alejandra; Sánchez, Federico; Martínez-Romero, Esperanza

    2016-01-01

    We present the complete genome sequence of Bradyrhizobium sp. strain CCGE-LA001, a nitrogen-fixing bacterium isolated from nodules of Phaseolus microcarpus. Strain CCGE-LA001 represents the first sequenced bradyrhizobial strain obtained from a wild Phaseolus sp. Its genome revealed a large and novel symbiotic island. PMID:26988045

  9. Screening diagnostic candidates for schistosomiasis from tegument proteins of adult Schistosoma japonicum using an immunoproteomic approach.

    Directory of Open Access Journals (Sweden)

    Min Zhang

    2015-02-01

    Full Text Available Schistosomiasis is one of the world's most prevalent zoonotic diseases and a serious worldwide public health problem. Since the tegument (TG of Schistosoma japonicum is in direct contact with the host and induces a host immune response against infection, the identification of immune response target molecules in the schistosome TG is crucial for screening diagnostic antigens for this disease.In this study, an immunoproteomics approach used TG proteins as screening antigens to identify potential diagnostic molecules of S. japonicum. Ten spots corresponding to six proteins were identified that immunoreacted with sera from S. japonicum-infected rabbits but not sera from uninfected rabbits and their specific IgG antibody levels declined quickly after praziquantel treatment. Recombinant phosphoglycerate mutase (PGM and UV excision repair protein RAD23 homolog B (RAD23 proteins were expressed and their diagnostic potential for schistosomiasis was evaluated and compared with schistosome soluble egg antigen (SEA using ELISA. The results showed high sensitivity and specificity and low crossreactivity when rSjPGM-ELISA and rSjRAD23-ELISA were used to detect water buffalo schistosomiasis. Moreover, antibodies to rSjPGM and rSjRAD23 might be short-lived since they declined quickly after chemotherapy.Therefore, the two schistosome TG proteins SjPGM and SjRAD23 were identified as potential diagnostic markers for the disease. The two recombinant proteins might have the potential to evaluate the effectiveness of drug treatments and for distinguishing between current and past infection.

  10. Physicochemical consequences of the perdeuteriation of glutathione S-transferase from S. japonicum

    OpenAIRE

    Brockwell, David; Yu, Lu; Cooper, Serena; Mccleland, Steven; Cooper, Alan; Attwood, David; Gaskell, Simon J.; Barber, Jill

    2001-01-01

    Glutathione S-transferase (GST) from Schistosoma japonicum has been prepared in both normal protiated (pGST) and fully deuteriated (dGST) form by recombinant DNA technology. Electrospray mass spectrometry showed that the level of deuteriation in dGST was 96% and was homogeneous across the sample. This result is attributed to the use of a deuterium-tolerant host Escherichia coli strain in the preparation of the protein. 10 heteroatom-bound deuteriums (in addition to the carbon-bound deuteriums...

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC-MS.

    Science.gov (United States)

    Ganzera, Markus; Pöcher, Astrid; Stuppner, Hermann

    2005-01-01

    The Chinese Pharmacopoeia indicates the use of field thistle (Cirsium setosum) and Japanese field thistle (C. japonicum) in the treatment of bleeding and inflammation. In the absence of an analytical method for the differentiation and analysis of these two species, TLC and HPLC-MS methods have been developed for this purpose. Both species could be readily distinguished by their flavonoid pattern as revealed by TLC on silica gel layers eluted with ethyl acetate:formic acid:acetic acid:water. The quantitative determination of four flavonoids, namely hispidulin-7-neohesperidoside, linarin, pectolinarin and luteolin, was possible using HPLC. Their optimum separation was achieved on a C12 column eluted with water and 0.025% trifluoroacetic acid in acetonitrile. HPLC-MS experiments were performed to confirm peak identity. In samples of C. japonicum, pectolinarin was the major flavonoid (0.32-2.00%), followed by linarin, hispidulin-7-neohesperidoside and luteolin; the total flavonoid content varied from 0.81 to 3.67%. In C. setosum only one flavonoid (linarin; 1.36-2.83%) was assignable. The HPLC method was validated for linearity, limit of detection (< or = 1.7 ng on-column), peak purity, repeatability (< or = 2.3%) and accuracy (recovery rates of spiked samples were between 99.2 and 101.6%).

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

    Directory of Open Access Journals (Sweden)

    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

  14. Cloning and expression in Escherichia coli of a new gene of Schistosoma japonicum encoding casein kinase Ⅱ beta subunit

    Institute of Scientific and Technical Information of China (English)

    彭寨玉; 余新炳; 吴忠道; 徐劲; 吴德; 李孜

    2004-01-01

    Background Nowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase Ⅱ beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E.coli).Methods The ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E.coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot. Results A 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase Ⅱ beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E.coli JM109.The recombinant protein could be recognized by the S. japonicum infected rabbit serum. Conclusion The full-length cDNA sequences encoding S. japonicum casein kinase Ⅱ beta subunit were firstly sequenced, cloned, and expressed in E.coli.

  15. O-Demethylation and successive oxidative dechlorination of methoxychlor by Bradyrhizobium sp. strain 17-4, isolated from river sediment.

    Science.gov (United States)

    Satsuma, Koji; Masuda, Minoru; Sato, Kiyoshi

    2012-08-01

    O-Demethylation of insecticide methoxychlor is well known as a phase I metabolic reaction in various eukaryotic organisms. Regarding prokaryotic organisms, however, no individual species involved in such reaction have been specified and characterized so far. Here we successfully isolated a bacterium that mediates oxidative transformation of methoxychlor, including O-demethylation and dechlorination, from river sediment. The isolate was found to be closely related to Bradyrhizobium elkanii at the 16S rRNA gene sequence level (100% identical). However, based on some differences in the physiological properties of this bacterium, we determined that it was actually a different species, Bradyrhizobium sp. strain 17-4. The isolate mediated O-demethylation of methoxychlor to yield a monophenolic derivative [Mono-OH; 1,1,1-trichloro-2-(4-hydroxyphenyl)-2-(4-methoxyphenyl)ethane] as the primary degradation product. The chiral high-performance liquid chromatography (HPLC) analysis revealed that the isolate possesses high enantioselectivity favoring the formation of (S)-Mono-OH (nearly 100%). Accompanied by the sequential O-demethylation to form the bis-phenolic derivative Bis-OH [1,1,1-trichloro-2,2-bis(4-hydroxyphenyl)ethane], oxidative dechlorination of the side chain proceeded, and monophenolic carboxylic acid accumulated, followed by the formation of multiple unidentified polar degradation products. The breakdown proceeded more rapidly when reductively dechlorinated (dichloro-form) methoxychlor was applied as the initial substrate. The resultant carboxylic acids and polar degradation products are likely further biodegraded by ubiquitous bacteria. The isolate possibly plays an important role for complete degradation (mineralization) of methoxychlor by providing the readily biodegradable substrates.

  16. The Differential Expression of Immune Genes between Water Buffalo and Yellow Cattle Determines Species-Specific Susceptibility to Schistosoma japonicum Infection

    OpenAIRE

    Jianmei Yang; Zhiqiang Fu; Yang Hong; Haiwei Wu; Yamei Jin; Chuangang Zhu; Hao Li; Ke Lu; Yaojun Shi; Chunxiu Yuan; Guofeng Cheng; Xingang Feng; Jinming Liu; Jiaojiao Lin

    2015-01-01

    Water buffalo are less susceptible to Schistosoma japonicum infection than yellow cattle. The factors that affect such differences in susceptibility remain unknown. A Bos taurus genome-wide gene chip was used to analyze gene expression profiles in the peripheral blood of water buffalo and yellow cattle pre- and post-infection with S. japonicum. This study showed that most of the identified differentially expressed genes (DEGs) between water buffalo and yellow cattle pre- and post-infection we...

  17. Microarray Analysis of Gene Expression Profiles of Schistosoma japonicum Derived from Less-Susceptible Host Water Buffalo and Susceptible Host Goat

    OpenAIRE

    Jianmei Yang; Yang Hong; Chunxiu Yuan; Zhiqiang Fu; Yaojun Shi; Min Zhang; Liuhong Shen; Yanhui Han; Chuangang Zhu; Hao Li; Ke Lu; Jinming Liu; Xingang Feng; Jiaojiao Lin

    2013-01-01

    BACKGROUND: Water buffalo and goats are natural hosts for S. japonicum in endemic areas of China. The susceptibility of these two hosts to schistosome infection is different, as water buffalo are less conducive to S. japonicum growth and development. To identify genes that may affect schistosome development and survival, we compared gene expression profiles of schistosomes derived from these two natural hosts using high-throughput microarray technology. RESULTS: The worm recovery rate was low...

  18. A deep analysis of the small non-coding RNA population in Schistosoma japonicum eggs.

    Directory of Open Access Journals (Sweden)

    Pengfei Cai

    Full Text Available BACKGROUND: Schistosoma japonicum is a parasitic flatworm that causes zoonotic schistosomiasis. The typical outcome of schistosomiasis is hepatic granuloma and fibrosis, which is primarily induced by soluble egg-derived antigens. Although schistosomal eggs represent an important pathogenic stage to the host, the biology of this critical stage is largely unknown. We previously investigated the expression profiles of sncRNAs during different developmental stages of this parasite. However, using small RNA extracted from egg-deposited liver tissues generated limited information about sncRNAs in eggs. Here, we characterized the complete small RNAome in this stage of the parasite after optimization of RNA purification. METHODOLOGY AND PRINCIPAL FINDINGS: A library, SjE, was constructed with the small RNA extracted from S. japonicum eggs and subjected to high-throughput sequencing. The data were depicted by comprehensive bioinformatic analysis to explore the expression features of sncRNAs in the egg stage. MicroRNAs accounted for about one quarter of the total small RNA population in this stage, with a strongly biased expression pattern of certain miRNA family members. Sja-miR-71, sja-miR-71-5p, and sja-miR-36-3p were suggested to play important roles in embryo development. A panel of transfer RNA fragments (tRFs precisely processed from the 5' end of mature tRNAs was identified for the first time, which represented a strong egg stage-biased expression. The tRNA-Ala derived small RNAs were the most highly expressed Sj-tRFs in eggs. Further, the expression of siRNAs from 29 types of well-defined transposable elements (TEs was observed to be relatively stable among different developmental stages. CONCLUSIONS AND SIGNIFICANCE: In this study, we characterized the sncRNA profile in the egg stage of S. japonicum. Featured expression of sncRNAs, especially the tRNA-derived small RNAs, was identified, which was further compared with that of other developmental

  19. Evaluation of immunoassays for the diagnosis of Schistosoma japonicum infection using archived sera.

    Directory of Open Access Journals (Sweden)

    Jing Xu

    Full Text Available BACKGROUND: With a national program initiated recently to reduce transmission of Schistosoma japonicum in the People's Republic of China (P.R. China, there is an urgent need for accessible, quality-assured diagnostics for case detection, surveillance, and program monitoring of chemotherapy efficacy and other control interventions in areas of low endemicity. We compared the performance of nine immunodiagnostic tests developed in P.R. China for detection of antibodies against S. japonicum and established their priority for further assessment in field settings. METHODOLOGY/PRINCIPAL FINDINGS: Using the Kato-Katz technique as the reference standard, 240 well-characterized archived serum specimens (100 positive and 140 negative were evaluated in nine immunological tests developed in P.R. China. The enzyme-linked immunoelectrotransfer blot assay (EITB, which uses an adult worm extract of S. japonicum, supplied by the Center of Disease Control and Prevention, USA, was also evaluated. The sensitivity and specificity of each test were determined and the reproducibility of each test was assessed by evaluating operator-to-operator and run-to-run variation. In addition the simplicity of use for the end-user was evaluated. All tests showed good sensitivities ranging from 92.0% (95% confidence interval (CI: 86.7-97.3% to 98.0% (95% CI: 95.3-100.0%. The test specificities varied from 70.0% (95% CI: 62.4-77.6% to 97.1% (95% CI: 94.4-99.9%. All tests showed excellent reproducibility with a discordant rate in the range of 0-10.0% for operator-to-operator variation and run-to-run variation. All tests, except one magnetic particle-based enzyme-linked immunosorbent assay, were found to be easy to use, especially the dot immunogold filtration assays. CONCLUSIONS/SIGNIFICANCE: Most evaluated tests had acceptable performance characteristics and could make an impact on the schistosomiasis control programs in P.R. China. Three tests with the highest sensitivity

  20. Characterization and expression of a novel cystatin gene from Schistosoma japonicum.

    Science.gov (United States)

    He, Baohua; Cai, Guobin; Ni, Yonghui; Li, Ying; Zong, Hongying; He, Li

    2011-08-01

    Cystatins are a family of cysteine protease inhibitors that play a crucial role in the immune evasion from their host and in the adaptation to host defence. Here, we isolated a full-length cDNA sequence inferred to encode a novel cystatin gene from a blood fluke, Schistosoma japonicum. The cDNA, designated SjCystatin, comprised an open reading frame (ORF) of 306 bp, and encoded 101 amino acids with a predicted molecular weight of 11.3 kDa. This predicted protein shared a significant degree of sequence identity with the type I cystatin (stefin) of Schistosoma mansoni and Homo sapiens. These proteins exhibited a typical cystatin topology, including the absence of disulfide bonds and three conserved catalytic motifs, Gly at the N-terminus (Gly(6)), Gln-X-Val-X-Gly motif (Q(49)VVAG(53)) and an LP pair at the C-terminus (L(76)P(77)). The SjCystatin gene spanned 376 bp and contained three exons. The positions of two introns were conserved between the cystatin genes of trematodes and their vertebrate hosts. Reverse transcription polymerase chain reaction confirmed the transcription of SjCystatin in the egg, schistosomula and adult stages of S. japonicum. The encoding ORF region was cloned into pET-28a (+) prokaryotic expression vector. After purification, the recombinant protein SjCystatin (recSjCystatin), expressed in Escherichia coli, was used to immunize animals and produce its specific polyclonal antibody. Western blot analysis revealed that the native SjCystatin was expressed in the egg and adult stages. The enzyme activity assay of the recSjCystatin showed that it inhibited the proteolytic activity of papain. SjCystatin protein was mainly localized on the miracidium within eggs. Immunohistochemistry revealed that SjCystatin mainly localized in the epithelial cells lining the gut as well as the tegument on the surface of adult worms. The conserved genomic DNA structure among cystatin homologues of trematode and their vertebrate host emphasized the characteristics

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

    International Nuclear Information System (INIS)

    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 μWatt.min/cm2). 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)

  2. Delayed tail loss during the invasion of mouse skin by cercariae of Schistosoma japonicum.

    Science.gov (United States)

    Wang, Ting; Fang, Zheng-Ming; Lei, Jia-Hui; Guan, Fei; Liu, Wen-Qi; Bartlett, Ann; Whitfield, Phil; Li, Yong-Long

    2012-02-01

    A traditional assumption is that schistosome cercariae lose their tails at the onset of penetration. It has, however, recently been demonstrated that, for Schistosoma mansoni, cercarial tails were not invariably being shed as penetration took place and a high proportion of tails entered human skin under experimental conditions. This phenomenon was termed delayed tail loss (DTL). In this paper, we report that DTL also happens with S. japonicum cercariae during penetration of mouse skin. It occurred at all cercarial densities tested, from as few as 10 cercariae/2·25 cm(2) of mouse skin up to 200 cercariae. Furthermore, it was demonstrated that there was a density-dependent increase in DTL as cercarial densities increased. No such density-dependent enhancement was shown for percentage attachment over the same cercarial density range.

  3. Polymorphic microsatellites in the human bloodfluke, Schistosoma japonicum, identified using a genomic resource

    Directory of Open Access Journals (Sweden)

    Spear Robert

    2011-02-01

    Full Text Available Abstract Re-emergence of schistosomiasis in regions of China where control programs have ceased requires development of molecular-genetic tools to track gene flow and assess genetic diversity of Schistosoma populations. We identified many microsatellite loci in the draft genome of Schistosoma japonicum using defined search criteria and selected a subset for further analysis. From an initial panel of 50 loci, 20 new microsatellites were selected for eventual optimization and application to a panel of worms from endemic areas. All but one of the selected microsatellites contain simple tri-nucleotide repeats. Moderate to high levels of polymorphism were detected. Numbers of alleles ranged from 6 to 14 and observed heterozygosity was always >0.6. The loci reported here will facilitate high resolution population-genetic studies on schistosomes in re-emergent foci.

  4. Screening and Primary Characterization of New Antigen Genes of Schistosoma Japonicum

    Institute of Scientific and Technical Information of China (English)

    王敏; 易新元; 李先平; 曾宪芳; 章洁

    2004-01-01

    To find Schistosoma japonicum(S.j) new antigen gene thus provide more useful vaccine candidates, the cDNA library of S. j adult worm was screened with sera of rabbits immtmized with the membrane antigens of Schistosoma japonicum hepato-portal schistosomula (SjHmAg). The positive clones were amplified by PCR and sequenced, then the sequences of clones were compared with all sequences in GenBank database using Blast process. The new clones were submitted to GenBank for accession numbers. Fifteen positive clones were obtained after three rounds of i mmunoscreening. The size of S. j cDNA fragments in positive clones ranged from 0.7 kb-3.0 kb after automatically excised with the helper phage. Sequence analysis revealed that partial sequence of clone M5 had significant homology with S.j mitochondria rnRNA, the other positive clones were new S.j genes. M2 clone sequence (GenBank accession number AF502579) was 730 bp long it had a 117 bp open reading frame (ORF). The sequence of M15 (GenBank accession number AF502582) has no transmembrane region and encodes 92 amino acids, and its protein contains a ferredoxins iron-sulfur binding region signature and two VWFC signal regions. The size of M1 ,M8, M9, M12( GenBank accession numbers: AF502578, AF502580, AF500622, AF502581 ) ranges from 402 bp to 766 bp. It concluded that the sera from rabbit immunized with SjHmAg could recognize S. j specific antigens molecules, and these antigens may induce the protective immunity against S.j infection.

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

    Directory of Open Access Journals (Sweden)

    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

  6. Local Antiglycan Antibody Responses to Skin Stage and Migratory Schistosomula of Schistosoma japonicum.

    Science.gov (United States)

    Smit, Cornelis H; Kies, Christiaan L; McWilliam, Hamish E G; Meeusen, Els N T; Hokke, Cornelis H; van Diepen, Angela

    2016-01-01

    Schistosomiasis is a tropical disease affecting over 230 million people worldwide. Although effective drug treatment is available, reinfections are common, and development of immunity is slow. Most antibodies raised during schistosome infection are directed against glycans, some of which are thought to be protective. Developing schistosomula are considered most vulnerable to immune attack, and better understanding of local antibody responses raised against glycans expressed by this life stage might reveal possible glycan vaccine candidates for future vaccine research. We used antibody-secreting cell (ASC) probes to characterize local antiglycan antibody responses against migrating Schistosoma japonicum schistosomula in different tissues of rats. Analysis by shotgun Schistosoma glycan microarray resulted in the identification of antiglycan antibody response patterns that reflected the migratory pathway of schistosomula. Antibodies raised by skin lymph node (LN) ASC probes mainly targeted N-glycans with terminal mannose residues, Galβ1-4GlcNAc (LacNAc) and Galβ1-4(Fucα1-3)GlcNAc (LeX). Also, responses to antigenic and schistosome-specific glycosphingolipid (GSL) glycans containing highly fucosylated GalNAcβ1-4(GlcNAcβ1)n stretches that are believed to be present at the parasite's surface constitutively upon transformation were found. Antibody targets recognized by lung LN ASC probes were mainly N-glycans presenting GalNAcβ1-4GlcNAc (LDN) and GlcNAc motifs. Surprisingly, antibodies against highly antigenic multifucosylated motifs of GSL glycans were not observed in lung LN ASC probes, indicating that these antigens are not expressed in lung stage schistosomula or are not appropriately exposed to induce immune responses locally. The local antiglycan responses observed in this study highlight the stage- and tissue-specific expression of antigenic parasite glycans and provide insights into glycan targets possibly involved in resistance to S. japonicum infection

  7. Vigna unguiculata is nodulated in Spain by endosymbionts of Genisteae legumes and by a new symbiovar (vignae) of the genus Bradyrhizobium.

    Science.gov (United States)

    Bejarano, Ana; Ramírez-Bahena, Martha-Helena; Velázquez, Encarna; Peix, Alvaro

    2014-10-01

    Vigna unguiculata was introduced into Europe from its distribution centre in Africa, and it is currently being cultivated in Mediterranean regions with adequate edapho-climatic conditions where the slow growing rhizobia nodulating this legume have not yet been studied. Previous studies based on rrs gene and ITS region analyses have shown that Bradyrhizobium yuanmingense and B. elkanii nodulated V. unguiculata in Africa, but these two species were not found in this study. Using the same phylogenetic markers it was shown that V. unguiculata, a legume from the tribe Phaseolae, was nodulated in Spain by two species of group I, B. cytisi and B. canariense, which are common endosymbionts of Genisteae in both Europe and Africa. These species have not been found to date in V. unguiculata nodules in its African distribution centres. All strains from Bradyrhizobium group I isolated in Spain belonged to the symbiovar genistearum, which is found at present only in Genisteae legumes in both Africa and Europe. V. unguiculata was also nodulated in Spain by a strain from Bradyrhizobium group II that belonged to a novel symbiovar (vignae). Some African V. unguiculata-nodulating strains also belonged to this proposed new symbiovar. PMID:24867807

  8. The effect-enhancing and toxicity-reducing activity of Hypericum japonicum Thunb. extract in murine liver cancer chemotherapy

    OpenAIRE

    ZHANG, HONG-BO; Lu, Ping; CAO, WEN-BO; Zhang, Zhen-Hua; MENG, XIANG-LEI

    2012-01-01

    Chinese herbs are potential sources of antitumor drugs with immunoregulatory activity and few adverse effects. In the present study, we investigated whether the Hypericum japonicum Thunb. (HJT) extract enhanced the efficacy of 5-fluorouracil (5-FU) treatment in murine liver tumor xenografts and reduced toxicity of chemotherapy in hepatoma H22-bearing mice. Tumor weight and inhibition rate, thymus and spleen indices, as well as white blood cell (WBC) count were calculated. The phagocytic funct...

  9. Global expression analysis revealed novel gender-specific gene expression features in the blood fluke parasite Schistosoma japonicum.

    Directory of Open Access Journals (Sweden)

    Xianyu Piao

    Full Text Available BACKGROUND: Schistosoma japonicum is one of the remarkable Platyhelminths that are endemic in China and Southeast Asian countries. The parasite is dioecious and can reside inside the host for many years. Rapid reproduction by producing large number of eggs and count-react host anti-parasite responses are the strategies that benefit long term survival of the parasite. Praziquantel is currently the only drug that is effective against the worms. Development of novel antiparasite reagents and immune-prevention measures rely on the deciphering of parasite biology. The decoding of the genomic sequence of the parasite has made it possible to dissect the functions of genes that govern the development of the parasite. In this study, the polyadenylated transcripts from male and female S. japonicum were isolated for deep sequencing and the sequences were systematically analysed. RESULTS: First, the number of genes actively expressed in the two sexes of S. japonicum was similar, but around 50% of genes were biased to either male or female in expression. Secondly, it was, at the first time, found that more than 50% of the coding region of the genome was transcribed from both strands. Among them, 65% of the genes had sense and their cognate antisense transcripts co-expressed, whereas 35% had inverse relationship between sense and antisense transcript abundance. Further, based on gene ontological analysis, more than 2,000 genes were functionally categorized and biological pathways that are differentially functional in male or female parasites were elucidated. CONCLUSIONS: Male and female schistosomal parasites differ in gene expression patterns, many metabolic and biological pathways have been identified in this study and genes differentially expressed in gender specific manner were presented. Importantly, more than 50% of the coding regions of the S. japonicum genome transcribed from both strands, antisense RNA-mediated gene regulation might play a critical

  10. Global Expression Analysis Revealed Novel Gender-Specific Gene Expression Features in the Blood Fluke Parasite Schistosoma japonicum

    OpenAIRE

    Xianyu Piao; Pengfei Cai; Shuai Liu; Nan Hou; Lili Hao; Fan Yang; Heng Wang; Jianwei Wang; Qi Jin; Qijun Chen

    2011-01-01

    BACKGROUND: Schistosoma japonicum is one of the remarkable Platyhelminths that are endemic in China and Southeast Asian countries. The parasite is dioecious and can reside inside the host for many years. Rapid reproduction by producing large number of eggs and count-react host anti-parasite responses are the strategies that benefit long term survival of the parasite. Praziquantel is currently the only drug that is effective against the worms. Development of novel antiparasite reagents and imm...

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

  13. Schistosoma japonicum in Samar, the Philippines: infection in dogs and rats as a possible risk factor for human infection.

    Science.gov (United States)

    Carabin, H; McGARVEY, S T; Sahlu, I; Tarafder, M R; Joseph, L; DE Andrade, B B; Balolong, E; Olveda, R

    2015-06-01

    The role that animals play in the transmission of Schistosoma japonicum to humans in the Philippines remains uncertain and prior studies have not included several species, adjustment for misclassification error and clustering, or used a cohort design. A cohort study of 2468 people providing stool samples at 12 months following praziquantel treatment in 50 villages of Western Samar, the Philippines, was conducted. Stool samples from dogs, cats, rats, and water buffaloes were collected at baseline (2003-2004) and follow-up (2005). Latent-class hierarchical Bayesian log-binomial models adjusting for misclassification errors in diagnostic tests were used. The village-level baseline and follow-up prevalences of cat, dog, and rat S. japonicum infection were associated with the 12-month cumulative incidence of human S. japonicum infection, with similar magnitude and precision of effect, but correlation between infection levels made it difficult to divide their respective effects. The cumulative incidence ratios associated with a 1% increase in the prevalence of infection in dogs at baseline and in rats at follow-up were 1·04 [95% Bayesian credible interval (BCI) 1·02-1·07] and 1·02 (95% BCI 1·01-1·04), respectively, when both species were entered in the model. Dogs appear to play a role in human schistosomiasis infection while rats could be used as schistosomiasis sentinels. PMID:25274409

  14. Adenovirus-mediated over-expression of Septin4 ameliorates hepatic fibrosis in mouse livers infected with Schistosoma japonicum.

    Science.gov (United States)

    He, Xue; Bao, Jing; Chen, Jinling; Sun, Xiaolei; Wang, Jianxin; Zhu, Dandan; Song, Ke; Peng, Wenxia; Xu, Tianhua; Duan, Yinong

    2015-12-01

    Septin4 (Sept4) belongs to Septin family and may be involved in apoptosis, vesicle trafficking and other cell processes. In this study, we attempted to investigate the effect of Sept4 in hepatic fibrosis induced by Schistosoma japonicum. ICR mice infected with S. japonicum for 12weeks were treated with PBS, Ad-ctr and Ad-Sept4, respectively. All mice were killed at 2weeks after injection, and the changes in the fibrotic livers were detected via H&E staining, Sirius red staining, qRT-PCR, western blot and TUNEL analysis. In addition, pcDNA3.1-Sept4 plasmid was transfected into LX-2 cells to observe the effect of Sept4 on apoptosis of HSCs in vitro. Ad-Sept4 could ameliorate liver fibrosis, as detected by H&E staining and Sirius red staining. The number of TUNEL-positive cells was increased in the Ad-Sept4 treated group. The expression of Sept4 and cleaved-caspase-3 were all augmented, while the expression of α-SMA, Col1α1 and IL-13 were reduced in the Ad-Sept4 treated group, compared with that expressed in the Ad-ctr group. Over-expression of Sept4 in LX-2 cells could promote apoptosis of LX-2 cells in vitro. In conclusion, Ad-Sept4 can attenuate the development of liver fibrosis induced by S. japonicum through apoptosis. PMID:26190030

  15. Local Auxin Biosynthesis Mediated by a YUCCA Flavin Monooxygenase Regulates Haustorium Development in the Parasitic Plant Phtheirospermum japonicum.

    Science.gov (United States)

    Ishida, Juliane K; Wakatake, Takanori; Yoshida, Satoko; Takebayashi, Yumiko; Kasahara, Hiroyuki; Wafula, Eric; dePamphilis, Claude W; Namba, Shigetou; Shirasu, Ken

    2016-08-01

    Parasitic plants in the Orobanchaceae cause serious agricultural problems worldwide. Parasitic plants develop a multicellular infectious organ called a haustorium after recognition of host-released signals. To understand the molecular events associated with host signal perception and haustorium development, we identified differentially regulated genes expressed during early haustorium development in the facultative parasite Phtheirospermum japonicum using a de novo assembled transcriptome and a customized microarray. Among the genes that were upregulated during early haustorium development, we identified YUC3, which encodes a functional YUCCA (YUC) flavin monooxygenase involved in auxin biosynthesis. YUC3 was specifically expressed in the epidermal cells around the host contact site at an early time point in haustorium formation. The spatio-temporal expression patterns of YUC3 coincided with those of the auxin response marker DR5, suggesting generation of auxin response maxima at the haustorium apex. Roots transformed with YUC3 knockdown constructs formed haustoria less frequently than nontransgenic roots. Moreover, ectopic expression of YUC3 at the root epidermal cells induced the formation of haustorium-like structures in transgenic P. japonicum roots. Our results suggest that expression of the auxin biosynthesis gene YUC3 at the epidermal cells near the contact site plays a pivotal role in haustorium formation in the root parasitic plant P. japonicum. PMID:27385817

  16. Local Auxin Biosynthesis Mediated by a YUCCA Flavin Monooxygenase Regulates Haustorium Development in the Parasitic Plant Phtheirospermum japonicum[OPEN

    Science.gov (United States)

    Takebayashi, Yumiko; Kasahara, Hiroyuki; Wafula, Eric; dePamphilis, Claude W.; Namba, Shigetou

    2016-01-01

    Parasitic plants in the Orobanchaceae cause serious agricultural problems worldwide. Parasitic plants develop a multicellular infectious organ called a haustorium after recognition of host-released signals. To understand the molecular events associated with host signal perception and haustorium development, we identified differentially regulated genes expressed during early haustorium development in the facultative parasite Phtheirospermum japonicum using a de novo assembled transcriptome and a customized microarray. Among the genes that were upregulated during early haustorium development, we identified YUC3, which encodes a functional YUCCA (YUC) flavin monooxygenase involved in auxin biosynthesis. YUC3 was specifically expressed in the epidermal cells around the host contact site at an early time point in haustorium formation. The spatio-temporal expression patterns of YUC3 coincided with those of the auxin response marker DR5, suggesting generation of auxin response maxima at the haustorium apex. Roots transformed with YUC3 knockdown constructs formed haustoria less frequently than nontransgenic roots. Moreover, ectopic expression of YUC3 at the root epidermal cells induced the formation of haustorium-like structures in transgenic P. japonicum roots. Our results suggest that expression of the auxin biosynthesis gene YUC3 at the epidermal cells near the contact site plays a pivotal role in haustorium formation in the root parasitic plant P. japonicum. PMID:27385817

  17. INFLUENCIA DE LA SEQUÍA SOBRE EL METABOLISMO DEL NITRÓGENO FIJADO DURANTE LA SIMBIOSIS Bradyrhizobium-SOYA

    Directory of Open Access Journals (Sweden)

    J. A. Freixas

    2010-01-01

    Full Text Available La soya (Glycine max L. Merr. establece simbiosis con bacterias fijadoras del nitrógeno de la familia de los rizobios; específicamente con bacterias del género Bradyrhizobium. Los rizobios secretan moléculas específicas denominadas factores Nod, que juegan un papel importante en el desarrollo del nódulo radicular. En el nódulo, los rizobios son diferenciados en bacteroides, donde ocurre la fijación biológica del nitrógeno (BNF y se produce amonio. La mayor parte del amonio es asimilado en glutamina, que participa indirectamente en la síntesis de ureidos. La soya se considera una de las plantas leguminosas más sensibles al estrés por sequía, con una disminución significativa en la BNF. Los ureidos se acumulan en plantas de soya sensibles a la sequía durante el déficit hídrico, mientras que las plantas tolerantes presentan bajas concentraciones de ureidos que pueden reducir el estrés sobre la BNF. Se han realizado investigaciones dirigidas a incrementar la BNF en condiciones de estrés por sequía. La fertilización con manganeso en condiciones moderadas de déficit hídrico incrementa la degradación de los ureidos y la BNF. La enzima ACC desaminasa en los rizobios degrada el ACC, precursor inmediato del etileno en las plantas, y disminuye los efectos inhibitorios del etileno en la nodulación. La inducción de los genes de la nodulación en Bradyrhizobium sp. ha mostrado efectos positivos en el crecimiento de la soya en condiciones moderadas de sequía. El objetivo de esta revisión bibliográfica está dirigido a relacionar nuevos blancos moleculares que permitan incrementar la BNF en condiciones de estrés por sequía.

  18. 东方田鼠抗日本血吸虫病机制研究%Study on the Mechanism of Microtus fortis Against Schistosoma Japonicum

    Institute of Scientific and Technical Information of China (English)

    刘金明; 刘瑞三; 林矫矫; 傅志强; 李浩; 魏梅雄; 孙军; 张亮; 何艳燕; 蒋守富

    2003-01-01

    This paper was about the study on the mechanism of Microtus fortis against Schistosoma japonicum. Firstly,we confirmed that Microtus fortis came from epidemic region (Dongting Lake beaches) and non-epidemic region (Qingtong Gorge in Ningxia province) were both resistant to Schistosoma japonicum infection after re-infection tests for several times. It seemed that their resistant ability was inheritable rather than acquired. Secondly, it was demonstrated by in vivo check-up and in vitro killing assay that there were some native antibodies of IgG3 subclass specifically to the schistosomula and adult worm of Schistosoma japonicum in Microtus fortis, which probably played an important role in resisting Schistosoma japonicum associated with complement. It was shown that macrophages and eosinophils in abdominal cavity of Microtus fortis had native ability of adhering to the schistosomula of Schistosoma japonicum. Then, the adult worm cDNA library of Schistosoma japonicum was screened with sera from Microtus fortis. Five positive clones were obtained, four of which were identified as new genes. Full-length cDNA of the two new genes were isolated by RACE. DNA vaccine was constructed with one named EST-mfs-3. After the Kunming mice immunized with this vaccine, the worm reduction rate and the egg reduction rate were 28.4% and 21.73% compared with that in control group respectively. This kind of DNA-based EST-mfs-3 vaccine was highly expressed in E. coli and induecd strong immune response in challenged group.Finally, two groups of cDNA probes prepared from liver and lung of Microtusfortis with or without Schistosoma japonicum infection were hybridized to the eDNA chip prepared from rat respectively. 156 and 332 genes revealed differential expression in infectious group compared with normal group. In conclusion, there would be many factors contribute to the mechanism of Microtusfortis against Schistosoma japonicum. We should stress the essentials and make further research on

  19. Effectiveness of synthetic trioxolane OZ78 against Schistosoma japonicum in mice and rabbits.

    Science.gov (United States)

    Xiao, Shu-Hua; Xue, Jian; Mei, Jing-Yan; Jiao, Pei-Ying

    2012-06-01

    Antischistosomal activities of a synthetic peroxide OZ78 (an ozonide carboxylic acid) against Schistosoma japonicum have been studied in mice and rabbits. Among 132 mice used, 30 of them were infected with 80-100 S. japonicum cercariae for collection of juvenile and adult schistosomes applied in in vitro tests. The remaining 102 mice were infected with 40 schistosome cercariae used for experimental treatment. Other 13 rabbits infected each with 200 schistosome cercariae were treated orally with OZ78 42 days post-infection. Most treated mice and rabbits were sacrificed 4 weeks post-treatment to collect residual schistosomes for evaluation of the drug efficacy. OZ78 and its sodium salt (OZ78-Na salt) 10-60 μg/mL alone exhibited no in vitro effect against day 14, day 21 schistosomula, and day 35 adult schistosomes. But OZ78 and OZ78-Na salt 10 and 20 μg/mL together with hemin 80 μg/mL showed decrease in worm motor activity and severe damage to the worm tegument and intestine, and all worms died within 3 days post-incubation. After infected mice were treated orally with OZ78 at a single dose of 400 mg/kg for 1 day, 34.9% of the worms shifted to the liver. Three and 7 days post-treatment, 100% of the worms were recovered from the liver. Fourteen days post-treatment, 92.3% of the worms still remained in the liver and 7.7% of the worms returned back to the mesenteric veins. Male and female worms shifted to the liver revealed in apparent shrinkage, degeneration of worm body, depigmentation in gut, and disappearance of ova in the uterus of some female worms. Meanwhile, dead worm and dead worm fragments were found in the liver tissues. In mice infected with various stages of schistosomes and treated orally with single OZ78 400 mg/kg, moderate or potential effect of the drug against day 0 (3-h-old worm), day 7, day 14, and day 21 juvenile worms and day 28, day 35 as well as day 42 adult worms were observed, the differences of total or female worm burdens between each

  20. Mitigation of soil N2O emission by inoculation with a mixed culture of indigenous Bradyrhizobium diazoefficiens

    Science.gov (United States)

    Akiyama, Hiroko; Hoshino, Yuko Takada; Itakura, Manabu; Shimomura, Yumi; Wang, Yong; Yamamoto, Akinori; Tago, Kanako; Nakajima, Yasuhiro; Minamisawa, Kiwamu; Hayatsu, Masahito

    2016-01-01

    Agricultural soil is the largest source of nitrous oxide (N2O), a greenhouse gas. Soybean is an important leguminous crop worldwide. Soybean hosts symbiotic nitrogen-fixing soil bacteria (rhizobia) in root nodules. In soybean ecosystems, N2O emissions often increase during decomposition of the root nodules. Our previous study showed that N2O reductase can be used to mitigate N2O emission from soybean fields during nodule decomposition by inoculation with nosZ++ strains [mutants with increased N2O reductase (N2OR) activity] of Bradyrhizobium diazoefficiens. Here, we show that N2O emission can be reduced at the field scale by inoculation with a mixed culture of indigenous nosZ+ strains of B. diazoefficiens USDA110 group isolated from Japanese agricultural fields. Our results also suggested that nodule nitrogen is the main source of N2O production during nodule decomposition. Isolating nosZ+ strains from local soybean fields would be more applicable and feasible for many soybean-producing countries than generating mutants. PMID:27633524

  1. Nodulation Characterization and Proteomic Profiling of Bradyrhizobium liaoningense CCBAU05525 in Response to Water-Soluble Humic Materials

    Science.gov (United States)

    Guo Gao, Tong; Yuan Xu, Yuan; Jiang, Feng; Zhen Li, Bao; Shui Yang, Jin; Tao Wang, En; Li Yuan, Hong

    2015-01-01

    The lignite biodegradation procedure to produce water-soluble humic materials (WSHM) with a Penicillium stain was established by previous studies in our laboratory. This study researched the effects of WSHM on the growth of Bradyrhizobium liaoningense CCBAU05525 and its nodulation on soybean. Results showed that WSHM enhanced the cell density of CCBAU05525 in culture, and increased the nodule number, nodule fresh weight and nitrogenase activity of the inoculated soybean plants. Then the chemical compounds of WSHM were analyzed and flavonoid analogues were identified in WSHM through tetramethyl ammonium hydroxide (TMAH)-py-GC/MS analysis. Protein expression profiles and nod gene expression of CCBAU05525 in response to WSHM or genistein were compared to illustrate the working mechanism of WSHM. The differently expressed proteins in response to WSHM were involved in nitrogen and carbon metabolism, nucleic acid metabolism, signaling, energy production and some transmembrane transports. WSHM was found more effective than genistein in inducing the nod gene expression. These results demonstrated that WSHM stimulated cell metabolism and nutrient transport, which resulted in increased cell density of CCBAU05525 and prepared the bacteria for better bacteroid development. Furthermore, WSHM had similar but superior functions to flavone in inducing nod gene and nitrogen fixation related proteins expression in CCBAU05525. PMID:26054030

  2. Mitigation of soil N2O emission by inoculation with a mixed culture of indigenous Bradyrhizobium diazoefficiens.

    Science.gov (United States)

    Akiyama, Hiroko; Hoshino, Yuko Takada; Itakura, Manabu; Shimomura, Yumi; Wang, Yong; Yamamoto, Akinori; Tago, Kanako; Nakajima, Yasuhiro; Minamisawa, Kiwamu; Hayatsu, Masahito

    2016-01-01

    Agricultural soil is the largest source of nitrous oxide (N2O), a greenhouse gas. Soybean is an important leguminous crop worldwide. Soybean hosts symbiotic nitrogen-fixing soil bacteria (rhizobia) in root nodules. In soybean ecosystems, N2O emissions often increase during decomposition of the root nodules. Our previous study showed that N2O reductase can be used to mitigate N2O emission from soybean fields during nodule decomposition by inoculation with nosZ++ strains [mutants with increased N2O reductase (N2OR) activity] of Bradyrhizobium diazoefficiens. Here, we show that N2O emission can be reduced at the field scale by inoculation with a mixed culture of indigenous nosZ+ strains of B. diazoefficiens USDA110 group isolated from Japanese agricultural fields. Our results also suggested that nodule nitrogen is the main source of N2O production during nodule decomposition. Isolating nosZ+ strains from local soybean fields would be more applicable and feasible for many soybean-producing countries than generating mutants. PMID:27633524

  3. Molluscicidal Activity of Nerium indicum Mill, Pterocarya stenoptera DC, and Rumex japonicum Houtt on Oncomelania hupensis

    Institute of Scientific and Technical Information of China (English)

    HONG WANG; WEI-MIN CAI; WAN-XIAN WANG; JIAN-MIN YANG

    2006-01-01

    To evaluate the molluscicidal activities of three Chinese plants N. Indicum Mill, P. Stenoptera DC, and R.japonicum Houtt, and to clarify the molluscicidal mechanism. Methods N-butanol extracts and water extracts of the three plants were obtained. The reactions of EST isozyme, glycogen and total protein of snails to the plant extracts were studied.Results EST electrophoresis showed that EST was an important antidotal enzyme system and reacted strongly to environment.EST changed greatly during the whole exposure period so that it could be viewed as a pathological index of toxicity. Extracts decreased the glycogen content of the snails'soft tissues greatly, and also the protein content. Conclusion All extracts show strong molluscicidal activity. The LD50 value of the water extract of N. Indicum Mill is as low as 13.2 mg/L. EST can be viewed as a pathological index of toxicity. The energy metabolism abnormity is the key reason for the molluscicidal activities. The biochemical mechanism needs further research.

  4. Expression of Recombinant Baculovirus Carrying Schistosoma japonicum 26 ku GST in Mammalian Cells

    Institute of Scientific and Technical Information of China (English)

    YU Guangqing; SONG Jianhua; LIU Wenqi; LONG Xiaochun; MO Hongmei; LI Yonglong; CHEN Xinwen

    2006-01-01

    In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into Tvector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac,then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 × 108. Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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.

  8. Schistosoma japonicum:Isolation and Identification of Peptides Mimicking Ferritin Epitopes from Phage Display Library

    Institute of Scientific and Technical Information of China (English)

    Lian-Fei TANG; Xin-Yuan YI; Xian-Fang ZENG; Lin-Qian WANG; Shun-Ke ZHANG

    2004-01-01

    In an attempt to isolate and identify the antigenic epitopes on ferritin of Schistosoma japonicum(SjFer)and to test their protective potentiality against Schistosomajaponicum(S.j),polyclonal antisera against SjFer was prepared to screen a 12-mer random peptide library.Three rounds of biopanning were performed and resulted in an enrichment.Six peptides selected randomly from the third elute were all found to be positive by evaluating the binding to anti-SjFer sera by ELISA and Western blotting.Three amino acid sequences were deduced from the six phage clones by sequencing.When they were used to vaccinate mice,the three peptides could induce significant reduction in adult worms(26.7% ,20.4%,and 25.9%)as well as in liver eggs per gram(LEPG)(40.0%,38.2%,and 40.8%).This result showed that three mimotopes on SjFer were obtained and they could induce significant protective efficacy against S.j.

  9. 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.

  10. Bradyrhizobium inoculation and nitrogen fertilization on cowpea grown in different soils = Inoculação com Bradyrhizobium e adubação nitrogenada em feijão-caupi cultivado em diferentes solos

    Directory of Open Access Journals (Sweden)

    Kelter Carvalho dos Santos

    2014-12-01

    Full Text Available Biological nitrogen fixation (BNF can be limited by the high availability of nitrogen (N. The soil clay fraction and organic matter content control the availability of N, and it is possible that these attributes affect the symbiotic fixation in soils fertilized with N. Based in that assumption, the objective with this study was to evaluate the effect of N fertilization on biological fixation in cowpea grown in five soils, with wide variation in their physical, chemical and and mineralogical attributes, representative of the growing areas of savannah of Roraima. The experiment was carried out in polyethylene pots in randomized blocks design with factorial arrangement 5x5, 5 soils (LAdx; LVd; PAdx; PVAd; RYve, 4 doses of nitrogen (0, 20,40, 80 kg ha-1 in inoculated plants and a control (without inoculation and nitrogen fertilization, with 4 replications. The seeds of BRS Guariba were inoculated with Bradyrhizobium strain BR 3262. The variables studied were: number, size and mass of the pod, root dry mass, nodule number and dry weight of nodules. The BNF provided an increase in the variables studied in all soils. N levels in inoculated plants provided significant increases varying between soil classes, 3–18% for yield components and 8-70% for the other variables. Nitrogen fertilization affects nodulation in cowpea, being restrictive in some soils. = A fixação biológica do nitrogênio (FBN pode ser limitada pela alta disponibilidade de nitrogênio (N. A fração argila e o teor de matéria orgânica do solo controlam a disponibilidade do N, sendo possível que esses atributos venham a interferir na fixação simbiótica em solos adubados com N. Com base nesse pressuposto, objetivou-se com o presente trabalho avaliar o efeito da adubação nitrogenada na fixação biológica do N em feijão-caupi cultivado em cinco solos, com ampla variação nos seus atributos físicos, químicos e mineralógicos, representativos das áreas de cultivo da Savana

  11. Influência da antibiose exercida por actinomicetos às estirpes de Bradyrhizobium SPP., na nodulação da soja Influence of antibiosis produced by actinomycetes on strains of Bradyrhizobium SPP. on soybean nodulation

    Directory of Open Access Journals (Sweden)

    João Carlos Pereira

    1999-01-01

    Full Text Available Este trabalho teve por objetivo avaliar o espectro antibiótico de actinomicetos provenientes de solos de Cerrados e a sua influência na nodulação da soja. As estirpes BR 29, BR 33, BR 40, BR 85, BR 86, BR 96, 47/587, 3B-7 e 4A-5 de Bradyrhizobium spp. apresentaram comportamento diferenciado em relação à resistência natural aos antibióticos produzidos por 204 actinomicetos. As estirpes BR 29 e BR 96 foram sensíveis a 5,2 e 9,9% dos antibióticos produzidos, respectivamente, enquanto a BR 33 apresentou sensibilidade a 20,3%. O antagonismo exercido pelos actinomicetos exclusivamente à BR 29 e BR 33 foi de 1,6 e 5,7%, respectivamente. Esse efeito não foi observado nas estirpes BR 40 e BR 96. Inoculações simples e em mistura das estirpes na presença de actinomicetos influenciaram a nodulação da soja. A co-inoculação da BR 33 e BR 29 com o isolado 370 reduziu o percentual de ocorrência média, nos nódulos, da BR 29, de 94,1% para 83,7%, com conseqüente aumento da BR 33 de 6,7% para 17,2%. Os resultados evidenciam a importância de estudos ecológicos desses microrganismos, visando avaliar o seu papel no estabelecimento de uma nodulação eficiente.The aim of this work was to evaluate the antibiotic spectrum of actinomycetes from Cerrado soils and their influence on soybean nodulation. Strains BR 29, BR 33, BR 40, BR 85, BR 86, BR 96, 47/587, 3B-7 and 4A-5 of Bradyrhizobium spp. were characterized by their natural resistence to antibiotics produced by 204 actinomycete isolates. The strains BR 29 and BR 96 of B. elkanii were sensitive to 5.2% and 9.9% the products of actinomycete isolates, respectively, while BR 33 was sensitive up to 20.3%. The antagonistic effects caused by actinomycete exclusively to BR 29 and BR 33 were 1.6% and 5.7% respectively. This effect was not observed for strains BR 40 and BR 96. Single and multistrains inoculations in the presence or absence of actinomycetes affected soybean nodulation. On double

  12. tfdA-Like Genes in 2,4-Dichlorophenoxyacetic Acid-Degrading Bacteria Belonging to the Bradyrhizobium-Agromonas-Nitrobacter-Afipia Cluster in α-Proteobacteria

    OpenAIRE

    Itoh, Kazuhito; Kanda, Rie; Sumita, Yoko; Kim, Hongik; Kamagata, Yoichi; Suyama, Kousuke; Yamamoto, Hiroki; Hausinger, Robert P.; Tiedje, James M.

    2002-01-01

    The 2,4-dichlorophenoxyacetate (2,4-D)/α-ketoglutarate dioxygenase gene (tfdA) homolog designated tfdAα was cloned and characterized from 2,4-D-degrading bacterial strain RD5-C2. This Japanese upland soil isolate belongs to the Bradyrhizobium-Agromonas-Nitrobacter-Afipia cluster in the α subdivision of the class Proteobacteria on the basis of its 16S ribosomal DNA sequence. Sequence analysis showed 56 to 60% identity of tfdAα to representative tfdA genes. A MalE-TfdAα fusion protein expressed...

  13. Construction of a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum and expression in Escherichia coli BL21( DE3)

    Institute of Scientific and Technical Information of China (English)

    张丽

    2014-01-01

    Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj)in Escherichia coli(E.coli)BL21(DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosoma japonicum(Sj26GST)antigen gene was amplified by real-time PCR(RT-PCR)from the total RNA,then cloned into a prokaryotic expression plasmid pGEX-1λT and transformed into E.coli BL21(DE3)to construct pGEX-Sj26GST;BL21(pGEX-

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Genetic variation between Schistosoma japonicum lineages from lake and mountainous regions in China revealed by resequencing whole genomes.

    Science.gov (United States)

    Yin, Mingbo; Liu, Xiao; Xu, Bin; Huang, Jian; Zheng, Qi; Yang, Zhong; Feng, Zheng; Han, Ze-Guang; Hu, Wei

    2016-09-01

    Schistosoma infection is a major cause of morbidity and mortality worldwide. Schistosomiasis japonica is endemic in mainland China along the Yangtze River, typically distributed in two geographical categories of lake and mountainous regions. Study on schistosome genetic diversity is of interest in respect of understanding parasite biology and transmission, and formulating control strategy. Certain genetic variations may be associated with adaptations to different ecological habitats. The aim of this study is to gain insight into Schistosoma japonicum genetic variation, evolutionary origin and associated causes of different geographic lineages through examining homozygous Single Nucleotide Polymorphisms (SNPs) based on resequenced genome data. We collected S. japonicum samples from four sites, three in the lake regions (LR) of mid-east (Guichi and Tonglin in Anhui province, Laogang in Hunan province) and one in mountainous region (MR) (Xichang in Sichuan province) of south-west of China, resequenced their genomes using Next Generation Sequencing (NGS) technology, and made use of the available database of S. japonicum draft genomic sequence as a reference in genome mapping. A total of 14,575 SNPs from 2059 genes were identified in the four lineages. Phylogenetic analysis confirmed significant genetic variation exhibited between the different geographical lineages, and further revealed that the MR Xichang lineage is phylogenetically closer to LR Guich lineage than to other two LR lineages, and the MR lineage might be evolved from LR lineages. More than two thirds of detected SNPs were nonsynonymous; functional annotation of the SNP-containing genes showed that they are involved mainly in biological processes such as signaling and response to stimuli. Notably, unique nonsynonymous SNP variations were detected in 66 genes of MR lineage, inferring possible genetic adaption to mountainous ecological condition. PMID:27207135

  16. Combined TLR7/8 and TLR9 ligands potentiate the activity of a Schistosoma japonicum DNA vaccine.

    Directory of Open Access Journals (Sweden)

    Xuefeng Wang

    Full Text Available BACKGROUND: Toll-like receptor (TLR ligands have been explored as vaccine adjuvants for tumor and virus immunotherapy, but few TLR ligands affecting schistosoma vaccines have been characterized. Previously, we developed a partially protective DNA vaccine encoding the 26-kDa glutathione S-transferase of Schistosoma japonicum (pVAX1-Sj26GST. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we evaluated a TLR7/8 ligand (R848 and a TLR9 ligand (CpG oligodeoxynucleotides, or CpG as adjuvants for pVAX1-Sj26GST and assessed their effects on the immune system and protection against S. japonicum. We show that combining CpG and R848 with pVAX1-Sj26GST immunization significantly increases splenocyte proliferation and IgG and IgG2a levels, decreases CD4(+CD25(+Foxp3(+ regulatory T cells (Treg frequency in vivo, and enhances protection against S. japonicum. CpG and R848 inhibited Treg-mediated immunosuppression, upregulated the production of interferon (IFN-γ, tumor necrosis factor (TNF-α, interleukin (IL-4, IL-10, IL-2, and IL-6, and decreased Foxp3 expression in vitro, which may contribute to prevent Treg suppression and conversion during vaccination and allow expansion of antigen-specific T cells against pathogens. CONCLUSIONS: Our data shows that selective TLR ligands can increase the protective efficacy of DNA vaccines against schistosomiasis, potentially through combined antagonism of Treg-mediated immunosuppression and conversion.

  17. Enhanced expression of the decoy receptor IL-13Rα2 in macrophages of Schistosoma japonicum-infected mice

    Institute of Scientific and Technical Information of China (English)

    WANG Wei; SHEN Yu-xian; LI Jing; ZHANG Shi-hai; LUO Qing-li; ZHONG Zhen-rong; JIANG Zuo-jun; SHEN Ji-long

    2009-01-01

    Background Type 2 cytokine interleukin (IL)-13 and its decoy receptor, IL-13 receptor (R)α2 appear to play a major role in tissue fibrosis of schistosomiasis and asthma. IL-13 is a key regulator of the extracellular matrix (ECM). It is known to signal to cells by binding to the IL-13Ra1, which then heterodimerizes with IL-4Rα. In contrast, IL-13Rα2 binds IL-13 with high affinity but does not signal. IL-13Rα2 is known to down-regulate granulomatous inflammation and prolong host survival in Schistosoma mansoni (S. Mansoni) infection, but little is known about the location and expression level of IL-13Ra2 in the context of S. Japonicum infection. Methods We established S. Japonicum-infected mouse models. Kinetic serum levels of IL-13Rα2 were examined with ELISA. IL-13Rα2 mRNA and protein of liver tissues were determined by PCR and immunoblotting analysis, respectively. Detection of IL-13Rα2 expression and location in macrophages was performed by TaqMan PCR and fluorescent immunocytochemistry technique, respectively. Results A marked elevation of mRNA and protein expression of IL-13Rα2 was observed in mice during S. Japonicum infection. An enhanced expression of IL-13Rg2 was further demonstrated in primary macrophages of murine schistosomiasis. Conclusions IL-13Rα2 in macrophages may be a critical contributor to pathogenesis of schistosomiasis. The data highlight the potential importance of cell signaling and antifibrotic gene therapeutics in T helper 2 cell (Th2)-mediated diseases.

  18. Comparative characterization of microRNAs in Schistosoma japonicum schistosomula from Wistar rats and BALB/c mice.

    Science.gov (United States)

    Han, Hongxiao; Peng, Jinbiao; Hong, Yang; Fu, Zhiqiang; Lu, Ke; Li, Hao; Zhu, Chuangang; Zhao, Qiuhua; Lin, Jiaojiao

    2015-07-01

    More than 40 kinds of mammals in China are known to be naturally infected with Schistosoma japonicum (S. japonicum) (Peng et al. Parasitol Res 106:967-76, 2010). Compared with permissive BALB/c mice, rats are less susceptible to S. japonicum infection and are considered to provide an unsuitable microenvironment for parasite growth and development. MicroRNAs (miRNAs), via the regulation of gene expression at the transcriptional and post-transcriptional levels, may be responsible for developmental differences between schistosomula in these two rodent hosts. Solexa deep-sequencing technology was used to identify differentially expressed miRNAs from schistosomula isolated from Wistar rats and BALB/c mice 10 days post-infection. The deep-sequencing analysis revealed that nearly 40 % of raw reads (10.37 and 10.84 million reads in schistosomula isolated from Wistar rats and BALB/c mice, respectively) can be mapped to selected mirs in miRBase or in species-specific genomes. Further analysis revealed that several miRNAs were differentially expressed in schistosomula isolated from these two rodents; 18 were downregulated (by 2-fold) (expression levels in rats compare with those in mice). Additionally, three novel miRNAs were primarily predicted and identified. Among the 41 differentially expressed miRNAs, 4 miRNAs had been identified with specific functions in schistosome development or host-parasite interaction, such as sexual maturation (sja-miR-1, sja-miR-7-5p), embryo development (sja-miR-36-3p) in schistosome, and pathogenesis of schistosomiasis (sja-bantam). Then, the target genes were mapped, filtered, and correlated with a set of genes that were differentially expressed genes in schistosomula isolated from mice and rats, which we identified in a S. japonicum oligonucleotide microarray analysis in a previous study. Gene Ontology (GO) analysis of the predicted target genes of 13 differentially expressed miRNAs revealed that they were involved in some important

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    Gretel Gómez

    2008-01-01

    celular. La influencia de las concentraciones de melaza y extracto acuoso de soya sobre la velocidad específica de crecimiento µ (h-1 de la cepa se estimó, variando las concentraciones de melaza en el medio Bradyfact de 3 a 20 g.L-1 y del extracto acuoso de soya de 5 a 20 g.L-1. Se obtuvieron las dinámicas de crecimiento para cada variante y se calculó la µ. Los resultados evidenciaron que concentraciones de melaza y extracto acuoso de soya superiores a 10,0 y 12,5 g.L-1, respectivamente, inhiben el crecimiento de Bradyrhizobium elkanii ICA 8001. Este fenómeno pudo ser explicado matemática y fisiológicamente mediante dos nuevos modelos cinéticos. Este trabajo contribuye al conocimiento de la fisiología de Bradyrhizobium, pues evidencia la relación entre la composición del medio de cultivo y los requerimientos nutricionales del microorganismo, así como el diseño del medio de cultivo puede garantizar la obtención de inoculantes más efectivos que contengan elevadas concentraciones celulares.

  1. 2-Cys peroxiredoxins from Schistosoma japonicum: the expression profile and localization in the life cycle.

    Science.gov (United States)

    Kumagai, Takashi; Osada, Yoshio; Kanazawa, Tamotsu

    2006-10-01

    Peroxiredoxin (Prx) is known to be an antioxidant protein that protects the organisms against various oxidative stresses and functions as a signal transductor. Here, we determined the full-length cDNA sequences of three types of Prx from an Asian blood fluke, Schistosoma japonicum: Prx-1, Prx-2 and Prx-3. According to the deduced amino acid sequences, only Prx-3 had a mitochondria-targeting sequence. Using RT-PCR, it was shown that these Prx genes were constitutively expressed in the eggs, cercariae and adult worms of the schistosome. Western blot analysis using antisera specific for each Prx revealed that all the three Prx proteins existed in these developmental stages. By immunolocalization analysis, Prx-1 existed on the surface of a miracidium and in the space between a miracidium and an eggshell. Furthermore, Prx-1 was deposited in the host tissues around the eggs. In adult worms, Prx-1 was not only expressed in the tegument, but also contained in their excretory/secretory products. The surface of the 7 day-schistosomula was stained with anti-Prx-1 antiserum. On the other hand, Prx-2 only existed inside the miracidia in eggs. In addition, Prx-2 was mainly detected in the sub-tegumental tissues, parenchyma, vitelline gland and gut epithelium of the adult worms, but was not detected in the tegument of adults and schistosomula. Taken together with previous reports by other investigators, these data suggest that Prx-1 acts to protect the parasite against the ROS produced by host immune cells, and that Prx-2 plays important roles in intracellular redox signaling and/or in the reduction of ROS generated through the hemoglobinolytic process in the digestive tract. PMID:16806527

  2. A Bayesian-based approach for spatio-temporal modeling of county level prevalence of Schistosoma japonicum infection in Jiangsu province, China.

    Science.gov (United States)

    Yang, Guo-Jing; Vounatsou, Penelope; Zhou, Xiao-Nong; Tanner, Marcel; Utzinger, Jürg

    2005-02-01

    Spatio-temporal variations of Schistosoma japonicum infection risk in Jiangsu province, China, were examined and the relationships between key climatic factors and infection prevalence at the county level were determined. The parasitological data were collected annually by means of cross-sectional surveys carried out in 47 counties from 1990 to 1998. Climatic factors, namely land surface temperature (LST) and normalized difference vegetation index (NDVI), were obtained from remote sensing satellite sensors. Bayesian spatio-temporal models were employed to analyze the data. The best fitting model showed that spatial autocorrelation in Jiangsu province decreased dramatically from 1990 to 1992 and increased gradually thereafter. A likely explanation of this finding arises from the large-scale administration of praziquantel for morbidity control of schistosomiasis. Our analysis suggested a negative association between NDVI and risk of S. japonicum infection. On the other hand, an increase in LST contributed to a significant increase in S. japonicum infection prevalence. We conclude that combining geographic information system, remote sensing and Bayesian-based statistical approaches facilitate integrated risk modeling of S. japonicum, which in turn is of relevance for allocation of scarce resources for control of schistosomiasis japonica in Jiangsu province and elsewhere in China, where the disease remains of public health and economic significance. PMID:15710436

  3. The Differential Expression of Immune Genes between Water Buffalo and Yellow Cattle Determines Species-Specific Susceptibility to Schistosoma japonicum Infection.

    Directory of Open Access Journals (Sweden)

    Jianmei Yang

    Full Text Available Water buffalo are less susceptible to Schistosoma japonicum infection than yellow cattle. The factors that affect such differences in susceptibility remain unknown. A Bos taurus genome-wide gene chip was used to analyze gene expression profiles in the peripheral blood of water buffalo and yellow cattle pre- and post-infection with S. japonicum. This study showed that most of the identified differentially expressed genes (DEGs between water buffalo and yellow cattle pre- and post-infection were involved in immune-related processes, and the expression level of immune genes was lower in water buffalo. The unique DEGs (390 in yellow cattle were mainly associated with inflammation pathways, while the unique DEGs (2,114 in water buffalo were mainly associated with immune-related factors. The 83 common DEGs may be the essential response genes during S. japonicum infection, the highest two gene ontology (GO functions were associated with the regulation of fibrinolysis. The pathway enrichment analysis showed that the DEGs constituted similar immune-related pathways pre- and post-infection between the two hosts. This first analysis of the transcriptional profiles of natural hosts has enabled us to gain new insights into the mechanisms that govern their susceptibility or resistance to S. japonicum infections.

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

    Directory of Open Access Journals (Sweden)

    Shuai Liu

    2014-10-01

    Full Text Available 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.

  5. Plastome sequences of Lygodium japonicum and Marsilea crenata reveal the genome organization transformation from basal ferns to core leptosporangiates.

    Science.gov (United States)

    Gao, Lei; Wang, Bo; Wang, Zhi-Wei; Zhou, Yuan; Su, Ying-Juan; Wang, Ting

    2013-01-01

    Previous studies have shown that core leptosporangiates, the most species-rich group of extant ferns (monilophytes), have a distinct plastid genome (plastome) organization pattern from basal fern lineages. However, the details of genome structure transformation from ancestral ferns to core leptosporangiates remain unclear because of limited plastome data available. Here, we have determined the complete chloroplast genome sequences of Lygodium japonicum (Lygodiaceae), a member of schizaeoid ferns (Schizaeales), and Marsilea crenata (Marsileaceae), a representative of heterosporous ferns (Salviniales). The two species represent the sister and the basal lineages of core leptosporangiates, respectively, for which the plastome sequences are currently unavailable. Comparative genomic analysis of all sequenced fern plastomes reveals that the gene order of L. japonicum plastome occupies an intermediate position between that of basal ferns and core leptosporangiates. The two exons of the fern ndhB gene have a unique pattern of intragenic copy number variances. Specifically, the substitution rate heterogeneity between the two exons is congruent with their copy number changes, confirming the constraint role that inverted repeats may play on the substitution rate of chloroplast gene sequences.

  6. iTRAQ-based comparative proteomic analysis of excretory-secretory proteins of schistosomula and adult worms of Schistosoma japonicum.

    Science.gov (United States)

    Cao, Xiaodan; Fu, Zhiqiang; Zhang, Min; Han, Yanhui; Han, Hongxiao; Han, Qian; Lu, Ke; Hong, Yang; Lin, Jiaojiao

    2016-04-14

    Schistosomiasis remains a serious public health problem with 200 million people infected and 779 million people at risk worldwide. The schistosomulum and adult worm are two stages of the complex lifecycle of Schistosoma japonicum and excretory/secretory proteins (ESPs) play a major role in host-parasite interactions. In this study, iTRAQ-coupled LC-MS/MS was used to investigate the proteome of ESPs obtained from schistosomula and adult worms of S. japonicum, and 298 differential ESPs were identified. Bioinformatics analysis of differential ESPs in the two developmental stages showed that 161 ESPs upregulated in schistosomula were associated with stress responses, carbohydrate metabolism and protein degradation, whereas ESPs upregulated in adult worms were mainly related to immunoregulation and purine metabolism. Recombinant heat shock protein 70 (HSP70) and thioredoxin peroxidase (TPx), two differential proteins identified in this study, were expressed. Further studies showed that rSjHSP70 and rSjTPx stimulated macrophages expressing high levels of the anti-inflammatory factors TGF-β, IL-10 and Arg-1, and suppressed the expression of the pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and iNOS in LPS-induced macrophages. This study provides new insights into the survival and development of schistosomes in the final host and helps identify vaccine candidates or new diagnostic reagents for schistosomiasis.

  7. Ultrastructural observation and gene expression profiling of Schistosoma japonicum derived from two natural reservoir hosts, water buffalo and yellow cattle.

    Directory of Open Access Journals (Sweden)

    Jianmei Yang

    Full Text Available Water buffalo and yellow cattle are the two of the most important natural reservoir hosts for Schistosoma japonicum in endemic areas of China, although their susceptibility differs, with water buffalo being less conducive to the growth and development of S. japonicum. Results from the current study show that the general morphology and ultrastructure of adult schistosomes derived from the two hosts also differed. Using high-throughput microarray technology, we also compared the gene expression profiles of adult schistosomes derived from the two hosts. We identified genes that were differentially expressed in worms from the two natural hosts. Further analysis revealed that genes associated with protein kinase and phosphatase, the stimulus response, and lipid and nucleotide metabolism were overexpressed, whereas genes associated with reproduction, anatomical structure morphogenesis and multifunctional motif were underexpressed in schistosomes from water buffalo. These differentially expressed genes were mainly involved in nucleotide, energy, lipid metabolism, energy metabolism, transcription, transport and signaling pathway. This suggests that they are key molecules affecting the survival and development of schistosomes in different natural host species. The results of this study add to current understanding of the interplay between parasites and their natural hosts, and provide valuable information for the screening of vaccine candidates or new drug targets against schistosomiasis in the natural reservoir hosts in endemic areas.

  8. 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.

  9. 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.

  10. 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.

  11. Immunodiagnostic efficacy of detection of Schistosoma japonicum human infections in China:a meta analysis

    Institute of Scientific and Technical Information of China (English)

    Wei Wang; Youzi Li; Hongjun Li; Yuntian Xing; Guoli Qu; Jianrong Dai; Yousheng Liang

    2012-01-01

    Objective: To assess the diagnostic efficacy of the currently most widely used indirect hemagglutination assay (IHA) and enzyme-linked immunosorbent assay (ELISA) for detection of Schistosoma japonicum human infections. Methods:A comprehensive search was undertaken from China National Knowledge Infrastructure, Wanfang Database, VIP Database, PubMed, Cochrane Library, Science Citation Index Expanded, Proquest, and the inclusion and exclusion criteria were strictly settled. The funnel plot was used to assess the publication bias, Cochran’s Q test was employed to measure the homogeneity between studies, a summary receiver operating characteristic (SROC) curve was used to compare the diagnostic accuracy between the IHA and ELISA qualitatively by means of the Weighted Least Square method, the Ordinary Least Square method and the Robust regression method, and the diagnostic odds ratio (DOR) was drawn to compare the accuracy quantitatively. Results:Out of 785 publications, 19 papers were eventually selected for analysis. Literature quality assessment indicated that minor publication bias existed in studies pertaining IHA test, but no bias was found in literatures regarding ELISA test. The heterogeneity test showed a heterogeneity between studies was present (χ2=466.07 and 34.67, both P values<0.0001). The areas under the SROC curves of IHA were all higher than that of ELISA test using the three methods (Weighted Least Square method:0.766 vs. 0.695, Ordinary Least Square method: 0.826 vs. 0.741, Robust regression: 0.815 vs. 0.715). The TPR*values for IHA and ELISA were 0.710, 0.759, 0.749, and 0.650, 0.686 and 0.666, respectively, and OR values were 5.997, 9.937, 8.893, and 3.432, 4.784 and 3.959, respectively. The DOR of IHA was 9.41 (95%CI:4.88-18.18), and 4.78 (95%CI:3.21-7.13) for ELISA. Conclusions:All above results revealed that the diagnostic performance of IHA is better than that of ELISA. However, taking into account their unsatisfactory diagnostic value in

  12. Animal Host of Schistosoma japonicum and Transmission of Schistosomiasis in Poyang Lake Region

    Institute of Scientific and Technical Information of China (English)

    林丹丹; 刘跃民; 胡飞; 张绍基

    2003-01-01

    本文从鄱阳湖区洲滩野粪污染指数、实验流行病学研究及耕牛放牧、尾蚴分析和钉螺感染的季节性变化等方面阐明了鄱阳湖区日本血吸虫的主要动物宿主为耕牛,病牛(尤其是 3岁以下耕牛)为当地血吸虫病的主要传染源;分析了鄱阳湖区血吸虫传播特征,每年春季(4~6月),为血吸虫在终宿主(人和牛)和中间宿主相互传播的"交汇点";涨水季节(7~8月)为血吸虫非易感季节,秋季(9~10月)为人畜感染的高峰,冬季(11月~翌年 3月),为洲滩枯水期,血吸虫传播休止.终宿主和中间宿主的易感区域均为村旁的洲滩.%The article clarified, according to the recent research results which have been done in Poyang Lake region of Jiangxi province, bovines are the most important animal hosts of Schistosoma japonicum and infected bovines are main infectious sources of Schistosomiasis in Poyang Lake region. Spring (March to June) is the " cross transmission stage" between definitive host and intermediate host, that is, spring is not only the susceptible season of infection for livestock and human, but also is the susceptible season of infection for snails. In flood season (July to August), the transmission of Schistosomiasis between livestock and snails belong to " low level phase" . Autumn (September to Octber) is the second seasonal peak of cercariae and is the susceptible season of infection for livestock and human. Winter(November to Feburary in next year) is the non- susceptible season of infection. Most of the susceptible zones of infection for livestock, human and snails are the marshlands near the endemic villages.

  13. CO-INOCULAÇÃO DE SEMENTES DE CAUPI COM Bradyrhizobium E Paenibacillus E SUA EFICIÊNCIA NA ABSORÇÃO DE CÁLCIO, FERRO E FÓSFORO PELA PLANTA CO-INOCULATION OF CAUPI SEEDS WITH Bradyrhizobium AND Paenibacillus AND ITS EFFICIENCY ON CALCIUM IRON AND PHOSPHORUS PLANT ABSORPTION

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo de Souza Fernandes da Silva

    2007-09-01

    Full Text Available

    O objetivo deste estudo foi verificar a viabilidade da co-inoculação de sementes de caupi (Vigna unguiculata (L. Walp. com bactérias fixadoras de nitrogênio dos gêneros Bradyrhizobium sp. (BR-2001 e NFB-700 e Paenibacillus polymyxa [(Loutit (L.]e sua eficiência na absorção de cálcio, ferro e fósforo pelas plantas de caupi, sob diferentes métodos de inoculação. Foi utilizado um Argissolo Amarelo coletado em fevereiro de 2002, localizado a BR 101 Norte, km 53, latitude 07º34'00'', longitude 35º00'00'' e altitude 14m, em Itapirema (Goiana, Estado de Pernambuco. As inoculações foram efetuadas na semente e no solo a uma profundidade de 3,5 cm, usando-se a cultivar IPA-205. Foram determinadas as concentrações de cálcio, ferro e fósforo na matéria seca da parte aérea das plantas de caupi. A co-inoculação do caupi com as estirpes de Bradyrhizobium sp. introduzidas no solo proporciona aumentos nas concentrações de cálcio, ferro e fósforo.

    PALAVRAS-CHAVE: Co-infecção; cálcio; ferro; fósforo; Vigna unguiculata.

    The objective of this study was to verify the viability of the co-inoculation of caupi seeds (Vigna unguiculata (L. Walp. with nitrogen fixing bacteria of Bradyrhizobium sp. (BR-2001 and NFB-700 and Paenibacillus polymyxa [(Loutit (L.] strains and their efficiency in calcium, iron, and phosphorus absorption by caupi plants under different inoculation methods. A Yellow Argisol was collected in February, 2002, located at the km 53 of BR 101 North highway, latitude 07ºSouth34'00'', longitude 35ºWest00'00'' and altitude 14m, in Itapirema (Goiana, Pernambuco State, Brazil. The inoculations were made in seeds and soil to a depth of 3.5 cm using IPA-205 cultivar. The variables evaluated were calcium, iron, and phosphorus concentration in the aerial

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

    Science.gov (United States)

    Sarr, Papa Saliou; Okon, Judith Wase; Begoude, Didier Aime Boyogueno; Araki, Shigeru; Ambang, Zachée; Shibata, Makoto; Funakawa, Shinya

    2016-01-01

    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 crop's biomass and represent a good strategy to improve soil fertility for subsequent cultivation. PMID:26904363

  15. Inoculação da soja com Bradyrhizobium no sulco de semeadura alternativamente à inoculação de sementes

    OpenAIRE

    Jerri Édson Zilli; Vicente Gianluppi; Rubens José Campo; Janaina Ribeiro Costa Rouws; Mariangela Hungria

    2010-01-01

    A fixação biológica de N é indispensável à soja no Brasil, dispensando o uso de N mineral e reduzindo o custo de produção. O objetivo deste estudo foi comparar a inoculação da soja com Bradyrhizobium no sulco de semeadura com a inoculação tradicional nas sementes, em solo com baixo teor de matéria orgânica e desprovido de bactérias nodulantes dessa cultura. Foi conduzido um experimento de campo no cerrado de Roraima no ano de 2006 e repetido nas mesmas condições em 2007, sendo avaliados os tr...

  16. Vigna mungo, V. radiata and V. unguiculata plants sampled in different agronomical-ecological-climatic regions of India are nodulated by Bradyrhizobium yuanmingense.

    Science.gov (United States)

    Appunu, Chinnaswamy; N'Zoue, Angèle; Moulin, Lionel; Depret, Géraldine; Laguerre, Gisèle

    2009-10-01

    Vigna mungo, Vigna radiata and Vigna unguiculata are important legume crops cultivated in India, but little is known about the genetic resources in native rhizobia that nodulate these species. To identify these bacteria, a core collection of 76 slow-growing isolates was built from root nodules of V. mungo, V. radiata and V. unguiculata plants grown at different sites within three agro-ecological-climatic regions of India. The genetic diversity of the bacterial collection was assessed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified DNA fragments of the 16S-23S rDNA intergenic spacer (IGS) region, and the symbiotic genes nifH and nodC. One rDNA IGS type grouped 91% of isolates, but more diversity was found at the symbiotic loci (17 symbiotic genotypes). Overall, no host plant specificity was shown, the three host plant species sharing common bradyrhizobial genotypes that represented 62% of the collection. Similarly, the predominant genotypes were found at most sampling sites and in all agro-ecological-climatic regions. Phylogenies inferred from IGS sequencing and multi-locus sequence analysis of the dnaK, glnII and recA genes indicated that all isolates but one were clustered with the Bradyrhizobium yuanmingense species. The nifH phylogeny also grouped the different nif haplotypes within a cluster including B. yuanmingense, except for one infrequent nif haplotype which formed a new lineage within the Bradyrhizobium genus. These results may reflect a long history of co-evolution between B. yuanmingense and Vigna spp. in India, while intra-species polymorphism detected in the symbiotic loci may be linked with the long history of diversification of B. yuanmingense coinciding with that of its host legumes.

  17. The dual actions of angiogenesis and anti-apoptosis induced by an isolated fraction from Geum japonicum repair muscle ischemia.

    Science.gov (United States)

    Cheung, Winghong L; Cheng, Lei; Liu, Hongwei; Gu, Xuemei; Li, Ming

    2007-03-01

    The fundamental improvement of muscle ischemia requires the re-establishment of sufficient vessel network. Despite many kinds of drugs have been used for ischemia, effective angiogenic drug is very limited. Here, we reported the identification and isolation of a potent angiogenic fraction (angio-T) from Geum japonicum and assessment of its therapeutic effects on muscle ischemia by reconstituting the insufficient blood supply network and enhancing cell survival potential. It was demonstrated that angio-T not only significantly enhanced the proliferation of cultured HCAECs in vitro, but also significantly enhanced the survival potential of the myofibers at risk and neovascularization in ischemic muscles leading to reconstitution of these vessel networks, significant reduction of ischemic areas, and significant myofiber regeneration in ischemic area one week post-ischemia.

  18. A rapid and simple method of detection of Blepharisma japonicum using PCR and immobilisation on FTA paper

    Directory of Open Access Journals (Sweden)

    Hughes Jacqueline M

    2003-09-01

    Full Text Available Abstract Background The rapid expansion in the availability of genome and DNA sequence information has opened up new possibilities for the development of methods for detecting free-living protozoa in environmental samples. The protozoan Blepharisma japonicum was used to investigate a rapid and simple detection system based on polymerase chain reaction amplification (PCR from organisms immobilised on FTA paper. Results Using primers designed from the α-tubulin genes of Blepharisma, specific and sensitive detection to the equivalent of a single Blepharisma cell could be achieved. Similar detection levels were found using water samples, containing Blepharisma, which were dried onto Whatman FTA paper. Conclusion This system has potential as a sensitive convenient detection system for Blepharisma and could be applied to other protozoan organisms.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Cloning, expression and characterisation of a type II cystatin from Schistosoma japonicum, which could regulate macrophage activation.

    Science.gov (United States)

    Yang, Xiao; Liu, Ju; Yue, Yuan; Chen, Wei; Song, Man; Zhan, Ximei; Wu, Zhongkai

    2014-11-01

    Cystatin play an important role in parasite immune evasion. It is involved in many immune responses processes regulations such as inhibiting antigen presentation, modifying cytokines production and macrophage polarization. In recent years, more and more cystatins were used in treating some inflammatory diseases such as asthma and inflammation bowel diseases; however, cystatins from Schistosoma japonicum were rarely studied. In the present study, we have cloned a cystatin from the adult stage of Schistosoma japonicum, named as SjCystatin, and its sequence shares conserved domains with other type II family cystatins. It was further verified by enzyme inhibition assays. SjCystatin retained its inhibitory activity under a wide range of pH values and temperatures, can maintain its inhibitory activity at pH 6.5-7.5 and 37 °C, respectively. Then, we investigated the effects of SjCystatin on the lipopolysaccharide (LPS)-induced activated RAW264.7. Results showed that SjCystatin inhibit LPS-induced nitric oxide production in a dose-dependent manner. LPS-induced TNF-α and IL-6 production began to be inhibited at least 6 h after SjCystatin stimulation. SjCystatin significantly increased IL-10 production at 6 h after stimulation and its effect on IL-10 production diminished quickly. These results imply that SjCystatin can induce M2 macrophage polarization and can be expected to serve as a potential drug source for the medication of inflammatory disorders like other cystatins.

  2. Distinct genetic diversity of Oncomelania hupensis, intermediate host of Schistosoma japonicum in mainland China as revealed by ITS sequences.

    Directory of Open Access Journals (Sweden)

    Qin Ping Zhao

    Full Text Available BACKGROUND: Oncomelania hupensis is the unique intermediate host of Schistosoma japonicum, which causes schistosomiasis endemic in the Far East, and especially in mainland China. O. hupensis largely determines the parasite's geographical range. How O. hupensis's genetic diversity is distributed geographically in mainland China has never been well examined with DNA sequence data. METHODOLOGY/PRINCIPAL FINDINGS: In this study we investigate the genetic variation among O. hupensis from different geographical origins using the combined complete internal transcribed spacer 1 (ITS1 and ITS2 regions of nuclear ribosomal DNA. 165 O. hupensis isolates were obtained in 29 localities from 7 provinces across mainland China: lake/marshland and hill regions in Anhui, Hubei, Hunan, Jiangxi and Jiangsu provinces, located along the middle and lower reaches of Yangtze River, and mountainous regions in Sichuan and Yunnan provinces. Phylogenetic and haplotype network analyses showed distinct genetic diversity and no shared haplotypes between populations from lake/marshland regions of the middle and lower reaches of the Yangtze River and populations from mountainous regions of Sichuan and Yunnan provinces. The genetic distance between these two groups is up to 0.81 based on Fst, and branch time was estimated as 2-6 Ma. As revealed in the phylogenetic tree, snails from Sichuan and Yunnan provinces were also clustered separately. Geographical separation appears to be an important factor accounting for the diversification of the two groups of O. hupensis in mainland China, and probably for the separate clades between snails from Sichuan and Yunnan provinces. In lake/marshland and hill regions along the middle and lower reaches of the Yangtze River, three clades were identified in the phylogenetic tree, but without any obvious clustering of snails from different provinces. CONCLUSIONS: O. hupensis in mainland China may have considerable genetic diversity, and a more

  3. 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

    Directory of Open Access Journals (Sweden)

    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

  4. 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

    OpenAIRE

    Qiu Chunhui; Liu Shengfa; Hong Yang; Fu Zhiqiang; Wei Meimei; Ai Dezhou; Lin Jiaojiao

    2012-01-01

    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 level...

  5. Microarray analysis of gene expression profiles of Schistosoma japonicum derived from less-susceptible host water buffalo and susceptible host goat.

    Directory of Open Access Journals (Sweden)

    Jianmei Yang

    Full Text Available BACKGROUND: Water buffalo and goats are natural hosts for S. japonicum in endemic areas of China. The susceptibility of these two hosts to schistosome infection is different, as water buffalo are less conducive to S. japonicum growth and development. To identify genes that may affect schistosome development and survival, we compared gene expression profiles of schistosomes derived from these two natural hosts using high-throughput microarray technology. RESULTS: The worm recovery rate was lower and the length and width of worms from water buffalo were smaller compared to those from goats following S. japonicum infection for 7 weeks. Besides obvious morphological difference between the schistosomes derived from the two hosts, differences were also observed by scanning and transmission electron microscopy. Microarray analysis showed differentially expressed gene patterns for parasites from the two hosts, which revealed that genes related to lipid and nucleotide metabolism, as well as protein folding, sorting, and degradation were upregulated, while others associated with signal transduction, endocrine function, development, immune function, endocytosis, and amino acid/carbohydrate/glycan metabolism were downregulated in schistosomes from water buffalo. KEGG pathway analysis deduced that the differentially expressed genes mainly involved lipid metabolism, the MAPK and ErbB signaling pathways, progesterone-mediated oocyte maturation, dorso-ventral axis formation, reproduction, and endocytosis, etc. CONCLUSION: The microarray gene analysis in schistosomes derived from water buffalo and goats provide a useful platform to disclose differences determining S. japonicum host compatibility to better understand the interplay between natural hosts and parasites, and identify schistosome target genes associated with susceptibility to screen vaccine candidates.

  6. Synthesis, Bioactivity Evaluation, and Toxicity Assessment of Novel Salicylanilide Ester Derivatives as Cercaricides against Schistosoma japonicum and Molluscicides against Oncomelania hupensis

    Science.gov (United States)

    Wang, Weisi; Qin, Zhiqiang; Zhu, Dan; Wei, Yufen; Li, Shizhu

    2015-01-01

    A series of novel salicylanilide ester derivatives were synthesized, characterized, and evaluated for cercaricidal potential against Schistosoma japonicum and molluscicidal potential against Oncomelania hupensis. Four derivatives exhibited remarkable cercaricidal activity superior to that of niclosamide. Among them, the most active compound, 4-chloro-2-((2-methoxy-4-nitrophenyl)carbamoyl)phenyl 4-methoxybenzoate (compound 4c), showed a marked minimum effective cercaricidal concentration as low as 0.43 μM and significant molluscicidal activity, with a 50% lethal concentration (LC50) of 0.206 g/m2. Particularly, compound 4c displayed 88-fold decreased fish toxicity on Danio rerio and 44-fold reduced cytotoxicity on human kidney HEK293 cells in comparison with the toxicity of niclosamide. The results indicated that 4c could serve as a promising drug candidate, with environmental safety properties, against Schistosoma japonicum at transmission stages. The preliminary molecular mechanism of target compounds in Schistosoma japonicum cercariae was also investigated. Salicylanilide ester derivatives exhibited an inhibitory effect on nitric oxide synthase (NOS) but no effect on lactate dehydrogenase (LDH) and acetylcholinesterase (AChE), and a strong and significant correlation between NOS inhibitory efficacy and cercaricidal activity was observed. In addition, 4c could downregulate the expression of NOS in a dose-dependent manner. These results suggested that NOS was probably one of the drug targets of salicylanilide esters. PMID:26503661

  7. Deep sequencing-based identification of pathogen-specific microRNAs in the plasma of rabbits infected with Schistosoma japonicum.

    Science.gov (United States)

    Cheng, Guofeng; Luo, Rong; Hu, Chao; Cao, Jie; Jin, Youxin

    2013-12-01

    Circulating microRNAs (miRNAs) have received considerable attention as a novel class of biomarkers for the diagnosis of cancer and as signalling molecules in mediating intercellular communication. Schistosomes, the causative agents of schistosomiasis, live in the blood vessels of a mammalian host in the adult stage. In the present study, we characterized schistosome-specific small RNA populations in the plasma of rabbits infected with Schistosoma japonicum (S. japonicum) using a deep sequencing method and then identified five schistosome-specific miRNAs, including four known miRNAs (Bantam, miR-3479, miR-10 and miR-3096), and one novel miRNA (miR-0001, miRBase ID: sja-miR-8185). Four of the five schistosome-specific miRNAs were also detected by real-time RT-PCR in the plasma of S. japonicum-infected mice. In addition, our study indicated that schistosome Argonaute 2/3 may be an excretory-secretory (ES) protein. In summary, our findings are expected to provide useful information for further development of novel biomarkers for the diagnosis of schistosomiasis and also for deeper understanding of the mechanism of host-parasite interaction.

  8. Comparison of apoptosis between adult worms of Schistosoma japonicum from susceptible (BALB/c mice) and less-susceptible (Wistar rats) hosts.

    Science.gov (United States)

    Wang, Tao; Guo, Xiaoyong; Hong, Yang; Han, Hongxiao; Cao, Xiaodan; Han, Yanhui; Zhang, Min; Wu, Miaoli; Fu, Zhiqiang; Lu, Ke; Li, Hao; Zhao, Zhixin; Lin, Jiaojiao

    2016-10-30

    Schistosomiasis remains a serious public health concern in China. BALB/c mice are susceptible to Schistosoma japonicum infection, whereas the Wistar rats are less susceptible. Apoptosis phenomenon was observed in 42d adult worms of S. japonicum from both rats and mice at the morphologic, DNA, cellular, and gene levels by transmission electron microscopy (TEM), fluorometric terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) analysis, fluorescein isothiocyanate-annexin-V/propidium iodide staining flow cytometry (FCM) analysis, and real-time PCR. The results showed that the apoptotic state in worms from two different susceptible hosts was diverse. Several classical hallmarks of apoptosis, including cell shrinkage, chromatin condensation and lunate marginalization, splitting of the nucleoli, nuclear shrinkage and apoptotic body formation were observed by TEM. TUNEL analysis showed that there were much more apoptosis spots in adult worms from rats than those from mice. Statistical analysis revealed that the degree of apoptosis and percentage of necrotic cells in adult worms from Wistar rats were significantly greater (Pworms from Wistar rats, as compared to those from BALB/c mice. The results obtained in this study collectively demonstrated that differential development of adult S. japonicum in less-susceptible rats and susceptible mice was significantly associated with apoptosis in the worm, and provided valuable information to guide further investigations of the mechanisms governing apoptosis and host interactions in schistosome infection.

  9. Effects of NaCl Stress on Chlorophyll Fluorescence Parameters in Cinnamomum japonicum var. chenii%NaCl胁迫对普陀樟叶绿素荧光参数的影响

    Institute of Scientific and Technical Information of China (English)

    李影丽; 汪奎宏; 杜国坚; 许利群; 杨华; 肖纪军

    2008-01-01

    [Objective] Study on the changes of chlorophyll fluorescence parameters in Cinnamomum japonicum var. chenii under NaCl stress. [Method] The seedling growth increment, chlorophyll content and chlorophyll fluorescence parameters in leaves of 1-year old Cinnamomum japonicum var. chenii were investigated in field experiment. [Result] Under NaCl stress, seedling growth increment reduced and the chlorophyll content decreased to a stable value; changes of Fv/Fm and Fv/Fo showed identical increasing trend and double peak type. With the aggravation of salt stress, most variations were ob- served in Fo, correlations among chlorophyll fluorescence parameters presented "rise-drop" trend (in the treatment of 7 g/L NaCl). [Condusion] Cinna-momum japonicum var. chenii is endowed with strong salt resistance and wide adaptability.

  10. Selection of Immunogenic Peptide Mimics of Male Worm Origin of Schistosoma Japonicum using Phage Display Peptide Library

    Institute of Scientific and Technical Information of China (English)

    陈欲晓; 易新元; 曾宪芳; 王林纤; 唐连飞; 章洁; McreynoldsLarry

    2004-01-01

    To select the immunogenic short peptide mimics of male worm origin of Schistosoma japonicum (Sj) and to explore their protection effect against schistosomiasis in mice, the random phage display peptide hbrary of 12 - mer was screened with IgG to soluble male worm antigen of Sj, and the specific positive clones selected through three rounds of screenings were detected by Dot-ELISA, and then injected subcutaneously into mice for vaccination and protection assessment against Sj. It was found that 18 randomly picked phage displayed clones all showed definite antigenicity with various intensities. The pooled phages displayed clones could induce production of specific antibodies and cause 31.72% of worm reduction rate and 51.54 % of egg reduction rate in mice, revealing a significant difference ( P < 0. 001 ) in comparison with those of the controls. It concludes that the short peptide mimics of male worm origin of Sj obtained by affinity screening phage display ptide library can elicit partial protection against this pathogen.

  11. DROUGHT-INDUCED EFFECTS AND RECOVERY OF NITRATE ASSIMILATION AND NODULE ACTIVITY IN COWPEA PLANTS INOCULATED WITH BRADYRHIZOBIUM SPP. UNDER MODERATE NITRATE LEVEL

    Directory of Open Access Journals (Sweden)

    Silveira Joaquim Albenísio Gomes da

    2001-01-01

    Full Text Available This study was carried out to establish comparative effects of drought and recovery on the nitrate assimilation and nodule activity related to N2 fixation in cowpea plants [Vigna unguiculata L. (Walp.] previously inoculated with Bradyrhizobium spp. BR-3256 (CB-756 strain in the presence of 5 mol m-3 NO-3. Twenty-eight-day-old nodulated plants were submitted to water deprivation during 4 consecutive days and afterwards resupplied with nutrient solution during 2 days. The water deprivation caused a rapid increase in the nitrate content in root and a marked reduction in leaf nitrate reductase (NR activity. In contrast nodule NR activity was slightly increased by water deprivation. Concomitantly, in nodules of water stressed plants, leghemoglobin and glutamine synthetase (GS activity declined and a progressive reduction in ureide-N concentration in xylem sap was observed. Leaf-NR activity increased rapidly after rehydration while leaf nitrate content declined. In contrast both GS activity and soluble protein content in the nodule continued to decline in rewatered plants. In addition the concentration of leghemoglobin recovered well, while the xylem ureide-N content experienced a slight increase after rehydration. Despite the nitrate assimilation in leaves and the nodule activity had been both severely affected by water stress, the rapid recovery of nitrate reductase activity suggests that the nitrate assimilation process is less sensitive to drought/rehydration cycle when cowpea plants are nodulated in presence of moderate nitrate level.

  12. 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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  14. Cloning, expression and protective immunity evalua- tion of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3′ and 5′ ends of the incomplete expression sequence tag (EST) of succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2 pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokary- otic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (posi- tive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction rate, the higher degree of egg reduction rate in the test group also sug- gested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.

  15. Cloning, expression and protective immunity evaluation of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    YU JunLong; WANG ShiPing; LI WenKai; DAI Gan; XU ShaoRui; HE Zhuo; PENG XianChu; ZHOU SongHua; LIU XueQin

    2007-01-01

    1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3' and 5' ends of the incomplete expression sequence tag (EST) of succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coll.SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (positive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction rate, the higher degree of egg reduction rate in the test group also suggested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.

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

    OpenAIRE

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

    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 (p...

  17. Cloning of three ZIP/Nramp transporter genes from a Ni hyperaccumulator plant thlaspi japonicum and their Ni2+ transport abilities

    OpenAIRE

    Mizuno, Takahumi; Usui, Koji; Horie, Kenji; Nosaka, Shiro; Mizuno, Naoharu; Obata, Hitoshi

    2005-01-01

    Ni homeostasis is essential for plant cell activity, but the mechanisms of Ni-transport and delivery are unknown. To elucidate the role of ZIP and NRAMP metal-transporters for Ni2+-transport and homeostasis, we cloned their homologous genes from the Ni hyperaccumulator Thlaspi japonicum, and investigated their Ni-transporting abilities by expression in yeast. The deduced amino acid sequences of the two Zip transporter genes (TjZnt1, TjZnt2) and one Nramp transporter gene cloned had high homol...

  18. Deep supercooling xylem parenchyma cells of katsura tree (Cercidiphyllum japonicum) contain flavonol glycosides exhibiting high anti-ice nucleation activity.

    Science.gov (United States)

    Kasuga, Jun; Hashidoko, Yasuyuki; Nishioka, Atsushi; Yoshiba, Megumi; Arakawa, Keita; Fujikawa, Seizo

    2008-09-01

    Xylem parenchyma cells (XPCs) of boreal hardwood species adapt to sub-freezing temperatures by deep supercooling to maintain a liquid state of intracellular water near -40 degrees C. Our previous study found that crude xylem extracts from such tree species exhibited anti-ice nucleation activity to promote supercooling of water. In the present study, thus, we attempted to identify the causative substances of supercooling. Crude xylem extracts from katsura tree (Cercidiphyllum japonicum), of which XPCs exhibited deep supercooling to -40 degrees C, were prepared by methanol extraction. The crude extracts were purified by liquid-liquid extraction and then by silica gel column chromatography. Although all the fractions obtained after each purification step exhibited some levels of anti-ice nucleation activity, only the most active fraction was retained to proceed to the subsequent level of purification. High-performance liquid chromatography (HPLC) analysis of a fraction with the highest level of activity revealed four peaks with high levels of anti-ice nucleation activity in the range of 2.8-9.0 degrees C. Ultraviolet (UV), mass and nuclear magnetic resonance (NMR) spectra revealed that these four peaks corresponded to quercetin-3-O-beta-glucoside (Q3G), kaempferol-7-O-beta-glucoside (K7G), 8-methoxykaempferol-3-O-beta-glucoside (8MK3G) and kaempferol-3-O-beta-glucoside (K3G). Microscopic observations confirmed the presence of flavonoids in cytoplasms of XPCs. These results suggest that diverse kinds of anti-ice nucleation substances, including flavonol glycosides, may have important roles in deep supercooling of XPCs. PMID:18518920

  19. 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...

  20. Estirpes de Bradyrhizobium em simbiose com guandu-anão em casa de vegetação e no campo

    Directory of Open Access Journals (Sweden)

    Márcia Rufini

    2014-03-01

    Full Text Available O objetivo deste trabalho foi avaliar a eficiência simbiótica de estirpes de bactérias fixadoras de nitrogênio do gênero Bradyrhizobium com guandu-anão. Os experimentos foram conduzidos em vasos de Leonard, em vasos com solo e em campo. Foram testadas 11 estirpes em vasos de Leonard, e as que apresentaram maior eficiência em promover o crescimento do guandu-anão foram avaliadas em vasos com solo (Latossolo Vermelho-Amarelo e Cambissolo e em campo (Latossolo Vermelho-Amarelo. Em todos os experimentos, os tratamentos foram comparados a dois controles positivos (estirpes aprovadas como inoculantes para as cultivares de guandu-anão BR 2003 e BR 2801 e a duas testemunhas sem inoculação, uma com alta concentração de N mineral, e a outra, a depender do experimento, sem N mineral (solo ou com baixa concentração de N (vasos de Leonard. Algumas estirpes proporcionaram crescimento vegetal semelhante ou superior às estirpes-referência e às testemunhas em vaso de Leonard. Em vasos com solo, o tipo de solo influenciou os tratamentos. No campo, não houve diferença entre os tratamentos, e as estirpes nativas promoveram bom crescimento. O guandu-anão é capaz de estabelecer associação simbiótica com bactérias fixadoras de N2, e a estirpe UFLA 03-320 apresenta potencial para ser recomendada para a cultura junto com a estirpe BR 2801.

  1. Unexpectedly Diverse Mesorhizobium Strains and Rhizobium leguminosarum Nodulate Native Legume Genera of New Zealand, while Introduced Legume Weeds Are Nodulated by Bradyrhizobium Species

    Science.gov (United States)

    Weir, Bevan S.; Turner, Susan J.; Silvester, Warwick B.; Park, Duck-Chul; Young, John M.

    2004-01-01

    The New Zealand native legume flora are represented by four genera, Sophora, Carmichaelia, Clianthus, and Montigena. The adventive flora of New Zealand contains several legume species introduced in the 19th century and now established as serious invasive weeds. Until now, nothing has been reported on the identification of the associated rhizobia of native or introduced legumes in New Zealand. The success of the introduced species may be due, at least in part, to the nature of their rhizobial symbioses. This study set out to address this issue by identifying rhizobial strains isolated from species of the four native legume genera and from the introduced weeds: Acacia spp. (wattles), Cytisus scoparius (broom), and Ulex europaeus (gorse). The identities of the isolates and their relationship to known rhizobia were established by comparative analysis of 16S ribosomal DNA, atpD, glnII, and recA gene sequences. Maximum-likelihood analysis of the resultant data partitioned the bacteria into three genera. Most isolates from native legumes aligned with the genus Mesorhizobium, either as members of named species or as putative novel species. The widespread distribution of strains from individual native legume genera across Mesorhizobium spp. contrasts with previous reports implying that bacterial species are specific to limited numbers of legume genera. In addition, four isolates were identified as Rhizobium leguminosarum. In contrast, all sequences from isolates from introduced weeds aligned with Bradyrhizobium species but formed clusters distinct from existing named species. These results show that native legume genera and these introduced legume genera do not have the same rhizobial populations. PMID:15466541

  2. The New Record of Trichogramma japonicum Ashmead,an Egg Parasitoid of Chilo suppressalis(Walker)in Jilin,China%吉林省稻螟赤眼蜂的发现和鉴定

    Institute of Scientific and Technical Information of China (English)

    郭震; 阮长春; 臧连生; 张帆; 靳锋云

    2011-01-01

    采用二化螟Chilo suppressalis(Walker)卵田间诱集的方法获得赤眼蜂Trichogramma寄生卵,室内培养羽化后,解剖并观察其雄性外生殖器特征,同时进行rDNA-ITS2序列分析,确定其为稻螟赤眼蜂T.japonicum Ashmead.BLAST分析结果表明,当地采集的供试赤眼蜂ITS2与广东的稻螟赤眼蜂相似性高达99.3%~100%.本研究首次确证吉林省存在寄生水稻二化螟的稻螟赤眼蜂自然种群.%Trichogramma wasps were collected using sentinel eggs of Chilo suppressalis (Walker) in rice fields. Throughmorphological identification of male genitalia and analysis of rDNA-ITS2 sequences, a Trichogramma species collectedwas identified as T. Japonicum Ashmead. The results of sequence alignment of ITS2 region showed that Trichogrammasp. Collected in Jilin had 99.3% - 100% similarity with T. Japonicum originated from Guangdong. The present workdemonstrated for the first time that there are natural populations of T. Japonicum parasitizing C. Suppressalis in Jilin,China.

  3. Effect of metal stress on photosynthetic pigments in the Cu-hyperaccumulating lichens Cladonia humilis and Stereocaulon japonicum growing in Cu-polluted sites in Japan.

    Science.gov (United States)

    Nakajima, Hiromitsu; Yamamoto, Yoshikazu; Yoshitani, Azusa; Itoh, Kiminori

    2013-11-01

    To understand the ecology and physiology of metal-accumulating lichens growing in Cu-polluted sites, we investigated lichens near temple and shrine buildings with Cu roofs in Japan and found that Stereocaulon japonicum Th. Fr. and Cladonia humilis (With.) J. R. Laundon grow in Cu-polluted sites. Metal concentrations in the lichen samples collected at some of these sites were determined by inductively coupled plasma mass spectroscopy (ICP-MS). UV-vis absorption spectra of pigments extracted from the lichen samples were measured, and the pigment concentrations were estimated from the spectral data using equations from the literature. Secondary metabolites extracted from the lichen samples were analyzed by high-performance liquid chromatography (HPLC) with a photodiode array detector. We found that S. japonicum and C. humilis are Cu-hyperaccumulating lichens. Differences in pigment concentrations and their absorption spectra were observed between the Cu-polluted and control samples of the 2 lichens. However, no correlation was found between Cu and pigment concentrations. We observed a positive correlation between Al and Fe concentrations and unexpectedly found high negative correlations between Al and pigment concentrations. This suggests that Al stress reduces pigment concentrations. The concentrations of secondary metabolites in C. humilis growing in the Cu-polluted sites agreed with those in C. humilis growing in the control sites. This indicates that the metabolite concentrations are independent of Cu stress. PMID:23953993

  4. Prevalence of Schistosoma japonicum infection of Oncomelania quadrasi snail colonies in 50 irrigated and rain-fed villages of Samar Province, the Philippines.

    Science.gov (United States)

    Madsen, Henry; Carabin, Hélène; Balolong, Don; Tallo, Veronica L; Olveda, Remigio; Yuan, M; McGarvey, Stephen T

    2008-03-01

    A cross-sectional survey of Oncomelania quadrasi, the intermediate host for Schistosoma japonicum, was conducted between 2004 and 2005 in 50 villages of the Province of Samar, the Philippines. The villages were classified as rain-fed (25) or with some man-made irrigation system (25). The primary objective was to identify all snail colony sites in the 50 villages and to compare snail population density and S. japonicum infection prevalence between the two types of villages. The presence of snail colonies was surveyed along streams, springs, various canals and swampy areas or grass land. A total of 198 colony sites were identified out of the 845 sites surveyed. Of these, a sufficient number of O. quadrasi snails were identified to measure density and infection in 147 sites. Density of O. quadrasi was remarkably uniform across habitats and there were no significant differences across habitats and between village type. The prevalence of infected snails showed more variability among habitats. Indeed, there was an interaction between the type of habitat and the type of village with irrigated villages being associated with a prevalence proportion ratio of 5.76 (1.31, 25.42) as compared to rain-fed villages among streams and springs. No such association was found among other habitats. The results suggest that once a suitable habitat exists, O. quadrasi populations establish and reach a plateau density. These results are discussed in light of possible ecological measures of control. PMID:18207119

  5. Gene Gun Bombardment with DNA-Coated Golden Particles Enhanced the Protective Effect of a DNA Vaccine Based on Thioredoxin Glutathione Reductase of Schistosoma japonicum

    Directory of Open Access Journals (Sweden)

    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.

  6. Gene Transcription Profile in Mice Vaccinated with Ultraviolet-attenuated Cercariae of Schistosoma japonicum Reveals Molecules Contributing to Elevated IFN-γLevels

    Institute of Scientific and Technical Information of China (English)

    Xiang ZHU; Feng LIU; Chuan SU; Guan-Ling WU; Zhao-Song ZHANG; Min-Jun JI; Hai-Wei WU; Yong WANG; Xiao-Ping CAI; Lei ZHANG; Shu-Ying HU; Lin-Lin FU

    2005-01-01

    Vaccination with ultraviolet-attenuated cercariae of Schistosoma japonicum induced protective immunity against challenge infection in experimental animal models. Our preliminary study on the transcription levels of IFN-γ and IL-4 in splenic CD4+ T cells revealed that attenuated cercariae elicited predominantly a Thl response in mice at the early stage, whereas normal cercariae stimulated primarily Th2dependent responses. Further analysis on the gene profile of the skin-draining lymph nodes demonstrated that the levels of IFN-γ were significantly higher in vaccinated mice than those in infected mice at day 4, 7 and 14 post-vaccination or post-infection. However, for IL-12 and IL-4, the potent inducers of Th l and Th2 responses, respectively, as well as IL-10, there were no differences over the course of the experiment between the infected and vaccinated mice. To explore the underlying factors that may potentially contribute to elevated IFN-γ in vaccinated mice, the mRNA profiles of the skin-draining lymph nodes at day 4 postexposure were compared using oligonucleotide microarrays. Within the 847 probe sets with increased signal values, we focused on chemokines, cytokines and relevant receptors, which were validated by semi-quantitative RT-PCR. A comprehensive understanding of the immune mechanisms of attenuated cercariae-induced protection may contribute to developing efficient vaccination strategies against S. japonicum, especially during the early stage of infection.

  7. Schistosoma japonicum:construction of phage display antibody library and its application in the immunodiagnosis of infection

    Institute of Scientific and Technical Information of China (English)

    陈代雄; 何蔼; 詹希美; 俞慕华; 雷智刚; 孟锦绣; 李卓雅; 梁瑜; 张瑞琳

    2004-01-01

    Background A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).Methods A phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.Results Six positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07×106 individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.Conclusions The results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.

  8. Establishment of AFLP Reaction System for Cypripedium japonicum Thunb.%扇脉杓兰AFLP反应体系的建立

    Institute of Scientific and Technical Information of China (English)

    李全健; 王彩霞; 田敏; 李翠新

    2012-01-01

    通过SDS法、高盐低pH法、常规CTAB法、高盐CTAB法和改良CTAB法提取扇脉杓兰(Cypripedium japonicum Thunb.)幼叶基因组DNA,并对其AFLP反应体系进行了优化.结果表明:改良CTAB法所提取的DNA电泳条带清晰无污染,A260和A280比值在1.8~2.0,用限制性内切酶酶切后条带均一,酶切时间以4h为宜.最终确定50 μL PCR反应体系的最佳条件为:预扩增产物稀释20倍4μL,dNTP(2.5 mmol/L)3 μL,Mg2+(25 mmol/L)4 μL,Mse I(10 μmol/L)和EcoR I(10 μmol/L)各1.5 μL,Taq DNA聚合酶(5 U/μL)0.3 μL.%Genomic DNA was extracted from young leaves of Cypripedium japonicum Thunb. By SDS method, high salt and low pH method, routine CTAB method, CTAB with high salt method and modified CTAB method, respectively, and an optimized AFLP reaction system was established for Cypripedium japonicum Thunb.. The results showed that DNA electrophoresis strips, which were extracted by the modified CTAB method, were clear and without contamination, and the proportion of A260 to A280 was from 1.8 to 2.0. After double digestion by restriction enzyme, the strips were uniform. 4 h is appropriate for enzyme restriction. The optimum PCR reaction system (50 μL) consisted of 4 μL 20 fold dilution of the pre-amplification product, 3 μL dNTPs (2.5 mmol/L), 4 (iL Mg2+ (25 mmol/L), 1.5 μL Mse I (10 μmol/L), 1.5 μL EcoR I (10 μmol/L) and 0.3 μL Taq DNA polymerase (5 U/μL)

  9. 日本血吸虫DNA疫苗联合免疫的研究进展%Research Progress on combined immunization of DNA vaccine against Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    冯其梅; 张树菊; 汪世平

    2012-01-01

    疫苗作为预防和控制传染病的重要手段,已成功的控制了很多传染病的流行.因此,许多研究者希望研制一种安全有效的血吸虫病疫苗,以期达到控制血吸虫病流行的目的.近年来,鉴于单价疫苗有限的免疫效果,人们开始进行日本血吸虫DNA疫苗联合免疫的研究.本文对日本血吸虫DNA疫苗联合免疫的研究资料进行了归纳整理,发现日本血吸虫DNA疫苗联合免疫涉及鸡尾酒式DNA混合疫苗、双价和多价DNA联合疫苗以及佐剂研究等方面,比较减虫卒、减卵率等指标,联合免疫疫苗的效果明显优于单价DNA疫苗的效果,而且具有潜在的实际应用价值.%As an important mean of prevention and control of infectious diseases, vaccine has been successfully took control. Therefore, in order to control the prevalence of sehistosomiasis japonica, many scientists want to develop a safe and effective vaccine against Schistosoma japonicum. Recently, in view of the limited immune effect of monovalent vaccine, people have started to study combined immunization of DNA vaccine of Schistosoma japonicum. This paper review the development of combined immunization of DNA vaccine against Schistosoma japonicum , and found that combined DNA vaccines include cocktail hybrid DNA vaccines, bivalent and multivalent DNA vaccines, and DNA vaccines with adjuvant mainly. They induced higher worm reduction rate and liver egg reduction rate, and would be well worthy of to be study.

  10. Saturation mutagenesis of Bradyrhizobium sp. BTAi1 toluene 4-monooxygenase at alpha-subunit residues proline 101, proline 103, and histidine 214 for regiospecific oxidation of aromatics.

    Science.gov (United States)

    Yanık-Yıldırım, K Cansu; Vardar-Schara, Gönül

    2014-11-01

    A novel toluene monooxygenase (TMO) six-gene cluster from Bradyrhizobium sp. BTAi1 having an overall 35, 36, and 38 % protein similarity with toluene o-xylene monooxygenase (ToMO) of Pseudomonas sp. OX1, toluene 4-monooxygenase (T4MO) of Pseudomonas mendocina KR1, and toluene-para-monooxygenase (TpMO) of Ralstonia pickettii PKO1, respectively, was cloned and expressed in Escherichia coli TG1, and its potential activity was investigated for aromatic hydroxylation and trichloroethylene (TCE) degradation. The natural substrate toluene was hydroxylated to p-cresol, indicating that the new toluene monooxygenase (T4MO·BTAi1) acts as a para hydroxylating enzyme, similar to T4MO and TpMO. Some shifts in regiospecific hydroxylations were observed compared to the other wild-type TMOs. For example, wild-type T4MO·BTAi1 formed catechol (88 %) and hydroquinone (12 %) from phenol, whereas all the other wild-type TMOs were reported to form only catechol. Furthermore, it was discovered that TG1 cells expressing wild-type T4MO·BTAi1 mineralized TCE at a rate of 0.67 ± 0.10 nmol Cl(-)/h/mg protein. Saturation and site directed mutagenesis were used to generate eight variants of T4MO·BTAi1 at alpha-subunit positions P101, P103, and H214: P101T/P103A, P101S, P101N/P103T, P101V, P103T, P101V/P103T, H214G, and H214G/D278N; by testing the substrates phenol, nitrobenzene, and naphthalene, positions P101 and P103 were found to influence the regiospecific oxidation of aromatics. For example, compared to wild type, variant P103T produced four fold more m-nitrophenol from nitrobenzene as well as produced mainly resorcinol (60 %) from phenol whereas wild-type T4MO·BTAi1 did not. Similarly, variants P101T/P103A and P101S synthesized more 2-naphthol and 2.3-fold and 1.6-fold less 1-naphthol from naphthalene, respectively.

  11. Vaccination of Goats with 31 kDa and 32 kDa Schistosoma japonicum Antigens by DNA Priming and Protein Boosting

    Institute of Scientific and Technical Information of China (English)

    Lianfei Tang; Zhijun Zhou; Yuxiao Chen; Yonghui Luo; Linqian Wang; Liyu Chen; Fushen Huang; Xianfang Zeng; Xinyuan Yi

    2007-01-01

    Two Schistosoma japonicum vaccine candidate antigens Sj 31 and Sj 32, which have shown particular promise to induce protective immunity in mice, were used to immunize goats by using a DNA priming-protein boosting strategy in present work. DNA vaccine formulations of the two antigens (VRSj31 and VRSj32) were produced and injected intramuscularly twice at a 2-week interval and then recombinant proteins (rSj31 and rSj32) together with Freund Complete Adjuvant (FCA) were used to boost the goats. The experiment was repeated in different batche cercariae. A strong anamnestic antibody response was induced after boost. A significant reduction of liver egg counts and miracidial hatching was showed in both experiments. Significant protections against challenge infection were elicited with 31.6% of percentage reduction for worm recovery in the second experiment and 20.9% in the first experiment, respectively.

  12. Effect of Verticillium lecanii on biological characteristics and life table of Serangium japonicum (Coleoptera: Coccinellidae), a predator of whiteflies under laboratory conditions

    Institute of Scientific and Technical Information of China (English)

    Lazreg Fatiha; Zhen Huang; Shun-Xiang Ren; Shaukat Ali

    2008-01-01

    Effects of entomopathogenic fungus Verticillium lecanii on biological characteristics and life table of Serangiumjaponicum, a predator of whiteflies against five different conidial concentrations (1×104, 1×105, 1×106, 1×107, and 1×108 conidia/mL) were studied under laboratory conditions. The developmental periods for all immature stages (from eggs, 1st, 2nd, 3rd, 4th instar nymph and pupae up to emergence) among the treatments were significantly different when compared to that of control, and the longest development period was observed as treated with 1×108spore/mL. However, no significant difference on the percent survival of all immature stages was observed among the treatments and control. Also, there were no significantly different effects of V. lecanii on mean generation time, intrinsic rate, the finite rate of increase and longevity of S. japonicum among the treatments and control.

  13. 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

    2016-02-01

    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

  14. Identification of Immunodominant Th1-type T cell Epitopes from Schistosoma japonicum 28 kDa Glutathione-S-transferase, a Vaccine Candidate

    Institute of Scientific and Technical Information of China (English)

    Guang-Fu LI; Guan-Ling WU; Yong WANG; Zhao-Song ZHANG; Xin-Jun WANG; Min-Jun JI; Xiang ZHU; Feng LIU; Xiao-Ping CAI; Hai-Wei WU

    2005-01-01

    Th1-type cytokines produced by the stimulation of Th1-type epitopes derived from defined schistosome-associated antigens are correlated with the development of resistance to the parasite infection.Schistosoma mansoni 28 kDa glutathione-S-transferase (Sm28GST), a major detoxification enzyme, has been recognized as a vaccine candidate and a phase Ⅱ clinical trial has been carried out. Sheep immunized with recombinant Schistosoma japonicum 28GST (Sj28GST) have shown immune protection against the parasite infection. In the present study, six candidate peptides (P1, P2, P3, P4, P7 and P8) from Sj28GST were predicted, using software, to be T cell epitopes, and peptides P5 and P6 were designed by extending five amino acids at the N-terminal and C-terminal of P1, respectively. The peptide 190-211 aa in Sj28GST corresponding to the Th1-type epitope (190-211 aa) identified from Sm28GST was selected and named P9.The nine candidate peptides were synthesized or produced as the fusion protein with thioredoxin in the pET32c(+)/BL21(DE3) system. Their capacity to induce a Th1-type response in vitro was measured using lymphocyte proliferation, cytokine detection experiments and flow cytometry. The results showed that P6(73-86 aa) generated the strongest stimulation effect on T cells among the nine candidate peptides, and drove the highest level of IFN-γ and IL-2. Therefore, P6 is a functional Th1-type T cell epitope that is different from that in Sm28GST, and will be useful for the development of effective vaccines which can trigger acquired immunity against S. japonicum. Moreover, our strategy of identifying the Th1-type epitope by a combination of software prediction and experimental confirmation provides a convenient and cost-saving alternative approach to previous methods.

  15. 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

  16. Schistosoma japonicum-infected hamsters (Mesocricetus auratus) used as a model in experimental chemotherapy with praziquantel, artemether, and OZ compounds.

    Science.gov (United States)

    Xiao, Shu-hua; Mei, Jing-yan; Jiao, Pei-ying

    2011-02-01

    The purpose of the study is to better understand the antischistosomal properties of artemether, praziquantel, and ozonide (OZ) compounds (synthetic trioxolanes, secondary ozonides) in hamster (Mesocricetus auratus) model. A total of 230 male hamsters infected each with 100 Schistosoma japonicum cercariae were used in the study. Groups of five to ten hamsters were treated orally with artemether, praziquantel, and OZ78 or OZ277 7-35 days post-infection at single doses of 50, 100, 150, or 200 mg/kg. Untreated but infected hamsters in each batch of test served as the control. All treated hamsters were sacrificed 4 weeks post-treatment for collection of residual worms using perfusion technique. Nonparametric method (Mann-Whitney test) was used to analyze the data. In groups of five hamsters treated with artemether 7, 14, 21, 28, and 35 days post-infection at single doses of 150 and 200 mg/kg, the difference of mean worm burden between each treated group and control group was statistically significant (P0.05). Further test with various single doses of 50-200 mg/kg confirmed the similar susceptibility of 7-day-old juvenile and 35-day-old adult schistosomes to artemether. After administration of praziquantel 100 mg/kg to groups of five hamsters 7, 21, and 35 days post-infection, higher worm burden reduction of 95.5% was seen in the group with 35-day-old adult schistosomes while in the groups with 7- and 21-day-old juvenile schistosomes, poor efficacy was seen with mean worm burden reductions of 36.6% and 35.6%. In the same batch of hamster treated with praziquantel 200 mg/kg, the moderate effect of the drug against 7- and 21-day-old worms was seen, but their mean worm burden was significantly higher than that of the group with adult schistosomes. In comparison of artemether and praziquantel against various stages of schistosomes, the results further demonstrated that artemether possessed similar effect against juvenile and adult schistosomes in hamsters, while praziquantel

  17. EVALUACIÓN DE LA INOCULACIÓN CONJUNTA Bradyrhizobium elkanii-HONGOS MICORRÍZICOSARBUSCULARES Y LA APLICACIÓN DE UN BIOESTIMULADOR DEL CRECIMIENTO VEGETAL EN SOYA CULTIVADA EN ÉPOCA DE INVIERNO

    Directory of Open Access Journals (Sweden)

    J. Corbera

    2010-01-01

    Full Text Available El estudio se desarrolló en el Instituto Nacional deCiencias Agrícolas (INCA sobre un suelo Ferralítico RojoLixiviado, con el objetivo de evaluar el efecto de la inoculacióncombinada de Bradyrhizobium elkanii y hongos micorrízicosarbusculares, junto con la aplicación de un bioestimulador delcrecimiento vegetal, sobre la respuesta del cultivar de soyaINCAsoy-24. Se empleó un diseño de bloques al azar con cuatroréplicas por tratamiento, los cuales consistieron en la inocula-ción de estos microorganismos, en sus formas simples y com-binados, mediante la tecnología de recubrimiento de semillas,así como la aplicación por diferentes vías del bioestimuladordel crecimiento vegetal a los tratamientos anteriores. Losresultados del estudio mostraron un efecto positivo de la ino-culación conjunta de ambos biofertilizantes sobre el crecimientoy rendimiento de este cultivar de soya, con incrementos de31,2 %, lo que corroboró los efectos sinérgicos y beneficiososde la coinoculación Bradyrhizobium elkanii-hongosmicorrízicos arbusculares en este cultivo. Los incrementos fue-ron ligeramente superiores con la aplicación a dicho tratamientodel estimulador del crecimiento vegetal, sin diferencias signifi-cativas con este, tanto recubriendo las semillas como con laaplicación foliar, con valores alrededor de 37,6 y 34,4 % respec-tivamente. Los tratamientos donde se aplicó el bioestimulanteprodujeron rendimientos superiores a sus controles donde nose aplicó el producto, obteniéndose incrementos en la producciónentre 2,44 y 14,91 %, de acuerdo al tratamiento evaluado, de-mostrando su efectividad en la estimulación del crecimientovegetal.

  18. EVALUACIÓN DE LA INOCULACIÓN CONJUNTA Bradyrhizobium elkanii-HONGOS MA Y LA APLICACIÓN DE UN BIOESTIMULADOR DEL CRECIMIENTO VEGETAL EN SOYA, CULTIVADA EN ÉPOCA DE PRIMAVERA

    Directory of Open Access Journals (Sweden)

    Jorge Corbera Gorotiza

    2011-01-01

    Full Text Available Se realizó un experimento en condiciones de camposobre un suelo Ferralítico Rojo Lixiviado, ubicado en las áreasdel Instituto Nacional de Ciencias Agrícolas, con el objetivode evaluar el efecto de la inoculación combinada deBradyrhizobium elkanii y una cepa de hongos MA, así comode la aplicación de un bioestimulador del crecimiento vegetal,sobre el crecimiento y rendimiento del cultivar de soyaINCAsoy-24, sembrado en época de primavera. Se empleó undiseño de bloques al azar con cuatro repeticiones portratamiento, que consistieron en la inoculación de losmicroorganismos, mediante la Tecnología de Recubrimientode Semillas, en sus formas simples y combinados, así como laaplicación por diferentes vías del bioestimulador del crecimientovegetal a los tratamientos anteriormente señalados. Los resultadosmostraron un efecto positivo de la coinoculación de ambosbiofertilizantes sobre el crecimiento y rendimiento de este cultivarde soya, con un incremento del rendimiento de 43,06 %, lo quedemuestra los efectos sinérgicos y beneficiosos de la aplicaciónconjunta Bradyrhizobium elkanii-hongos micorrízicosarbusculares en este cultivo. Los incrementos fueron ligeramentesuperiores con la aplicación a dicho tratamiento delbioestimulador del crecimiento vegetal, principalmente con elrecubrimiento de las semillas o con la aplicación foliar delproducto, los valores del incremento oscilaron entre 50,69 y55,56 % respectivamente. Los tratamientos donde fue aplicadoel bioestimulante produjeron rendimientos superiores a suscontroles donde no se aplicó el producto, obteniéndose incrementosen la producción entre 1,45 y 25,00 % de acuerdo al tratamientoevaluado, demostrándose efectividad del mismo en laestimulación del crecimiento vegetal.

  19. Biodegradace environmentálních polutantů Strukturní charakterizace nového typu halogenalkandehalogenasy LinB32 ze Sphingobium japonicum UT26

    OpenAIRE

    Němec, Pavel

    2013-01-01

    The main aim of this thesis was focused on protein crystallography and attempt to crystallize model protein Thaumatin and a new prepared haloalkane dehalogenase LinB32 of Sphingobium japonicum UT26. Crystallization trials were performed using the sitting and hanging drop vapour diffusion methods, crystallization under oil and crystallization in the capillaries. After measuring of diffraction data, the structure of LinB32 was modeled by the use of visualizing molecular modeling programs WinCoo...

  20. 叶巢外睾吸虫感染钉螺对钉螺体内日本血吸虫发育的影响%Studies on Antagonism between Exorchis ovariolobularis and Schistosoma japonicum in Oncomelania hupensis

    Institute of Scientific and Technical Information of China (English)

    叶向群

    2001-01-01

    目的探讨在日本血吸虫中间宿主湖北钉螺体内叶巢外睾吸虫和日本血吸虫的对抗性.方法通过叶巢外睾吸虫和日本血吸虫对湖北钉螺的双重感染,计算血吸虫的感染率和尾蚴逸出量,常规石蜡切片观察钉螺组织学变化.结果钉螺在感染血吸虫37 d后再感染外睾吸虫,经一定时间后检查发现,血吸虫的感染率为52.9%,显著低于同时单独感染日本血吸虫对照组的感染率(75.9%);先感染叶巢外睾吸虫,经10、32、60、100和120 d不同的时间间隔后再感染日本血吸虫的钉螺,血吸虫感染率分别为64%、66.7%、65.2%、56.4%和57.1%,而单独感染日本血吸虫对照组钉螺血吸虫感染率为90.5%,经统计检验,各双重感染实验组血吸虫与对照组的感染率间差异具有显著或非常显著性意义(P《0.05或P《0.01).对各试验组及对照组钉螺逸出尾蚴试验发现,试验组钉螺血吸虫尾蚴逸出量均显著低于对照组的逸出量.组织学观察发现各双重感染组钉螺消化腺萎缩,消化腺盲囊间隙只有少量血吸虫子胞蚴,血吸虫子胞蚴皱缩、变形及不规则,且子胞蚴中只含稀疏的尾蚴胚球,有的子胞蚴中已无胚球;而单独感染血吸虫的对照组中血吸虫均发育到成熟的子胞蚴或尾蚴.结论钉螺体内叶巢外睾吸虫和日本血吸虫之间存在一定的对抗性,而叶巢外睾吸虫在这种对抗中占优势.%Objective To examine the antagonistic interactions between Schistosoma japonicum and Exorchis ovariolobularis in the snail host, Oncomelania hupensis. Methods The infection rate and average number of cercariae of S.japonicum were examined in O. hupensis which had been suverinfected with S.japonicum and E. ovariolobularis. The experimental snails were histologically observed by paraffin section. Results When snails were fed with eggs of E. ovariolobularis 37 days later than S.japonicum infection, the infection rate of S .japonicum (52

  1. 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

    Directory of Open Access Journals (Sweden)

    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

  2. 野雉尾金粉蕨配子体发育及其系统学意义%Gametophyte development in Onychium japonicum and its systematic significance

    Institute of Scientific and Technical Information of China (English)

    邓晰朝

    2016-01-01

    Onychium japonicum falls into the category of Onychium Kaulf. of Sinopteridaceae;however, the alliance of Ony-chiumremains controversial. In this study, the spores of O. japonicum were artificially cultured in the original habitat humus soil and improved Knop′s nutrient solution in a constant temperature of 25℃ with 12 h of light and dark respectively per day and 2 500 lx of light intensity. Spore germination and gametophyte development were observed and recorded at every stage under the microscope. The results indicated that mature spores were tawny,tetrahedral,3-colporate,with fan-shaped e-quatorial view,perispore and reticulation on the surface of the exine. The germination procedure was Vittaria-type while the gametophyte development was Ceratopteris-type. The spores cultivated in either of the two substrates began to germinate a-bout 7 d and their germination shared the same Vittaria-type. With multiple transverse fission,the original gametophyte cell gradually turned into 3-9 protonemata with cylindrical shape, thin perispore,lumpy lateral exine and numerous granular chloroplasts. After about 15 d,the spores developed into prothallial plates and most of prothallial plates appeare spatulate.And 25 d later, young prothalliums were formed but they were not symmetrical;meanwhile, the gametophytes developed were Ceratopteris-type. When the prothalliums were developing, branching protonemata were flourishing and gametophytes were becoming fasciculate without any trichome. The rhizoids of O. japonicum, occasionally branching and lacking chloro-plasts, were unicellular and tubular. After about 45 d,antheridium, composed of a cap cell, a ring cell and a basal cell, e-merged from the edge of several irregular prothalliums. When the antheridium became mature, the cap cell split and sperm spilt. The antheridium looked like a circle from the top and an oval or a short column from the side. About 60 d later, a large archegonium was clearly visible. Its base was bigger

  3. 濒危植物扇脉杓兰的花部特征与繁育系统%Floral Characteristics and Breeding Systems of an Endangered Species Cypripedium japonicum

    Institute of Scientific and Technical Information of China (English)

    刘芬; 李全健; 王彩霞; 连静静; 田敏

    2013-01-01

    Cypripedium japonicum, a perennial herb of Orchidaceae, with unique flower shape, brilliant flower color, ornamental and medicinal value. Due to over-harvesting and the environment changes,a large number of wild populations of C. japonicum reduced, and some genetic resources lost. The species is in endangered, and listed as a national endangered plant. Consequently, it is essential to execute studies on floral characteristics and breeding systems of C. japonicum for understanding its extinct mechanism. We conducted a field investigation, and experiments of pollen germination test, emasculation, bagging and artificial pollination, and applied out-crossing index, to study the flowering character, pollen viability, stigma receptivity, breeding system of C. japonicum in natural populations. We also observed surface pattern of pollen and stigma by using an electron microscope. Results showed that: C. japonicum bloomed from April to June and the flowering span was 23 days among populations. The life span of one single flower was approximately 12-16 days. The stigma receptivity started from bloom and lasted for 14 days, and the pollen viability was high in the whole florescence. The stigma exceeded stamen in position and the relative difference in position between the stamen and style remained unchanged in the whole flowering process. The total value of OCI was 4. Seed setting propagation coefficient of C japonicum was low under field conditions, with only 5% . Based on the results of emasculation, bagging and artificial pollination experiments, C. japonicum is self-compatible and can't self-pollinate autokineticly and has no syngamy. Thus, the breeding system is pollinators depended.%通过野外观测,运用杂交指数(OCI),花粉萌发试验,去雄、套袋、人工授粉等方法对扇脉杓兰开花特性、花粉活力、柱头可授性、繁育系统进行研究,并对其柱头表面和花粉进行扫描电镜观察.结果表明:扇脉杓兰4-6

  4. A cross-sectional study of the prevalence of intensity of infection with Schistosoma japonicum in 50 irrigated and rain-fed villages in Samar Province, the Philippines

    Science.gov (United States)

    Tarafder, Mushfiqur R; Balolong, Ernesto; Carabin, Hélène; Bélisle, Patrick; Tallo, Veronica; Joseph, Lawrence; Alday, Portia; Gonzales, Ryan O'Neil; Riley, Steven; Olveda, Remigio; McGarvey, Stephen T

    2006-01-01

    Background Few studies have described heterogeneity in Schistosoma japonicum infection intensity, and none were done in Philippines. The purpose of this report is to describe the village-to-village variation in the prevalence of two levels of infection intensity across 50 villages of Samar Province, the Philippines. Methods This cross-sectional study was conducted in 25 rain-fed and 25 irrigated villages endemic for S. japonicum between August 2003 and November 2004. Villages were selected based on irrigation and farming criteria. A maximum of 35 eligible households were selected per village. Each participant was asked to provide stool samples on three consecutive days. All those who provided at least one stool sample were included in the analysis. A Bayesian three category outcome hierarchical cumulative logit regression model with adjustment for age, sex, occupation and measurement error of the Kato-Katz technique was used for analysis. Results A total of 1427 households and 6917 individuals agreed to participate in the study. A total of 5624 (81.3%) participants provided at least one stool sample. The prevalences of those lightly and at least moderately infected varied from 0% (95% Bayesian credible interval (BCI): 0%–3.1%) to 45.2% (95% BCI: 36.5%–53.9%) and 0% to 23.0% (95% BCI: 16.4%–31.2%) from village-to-village, respectively. Using the 0–7 year old group as a reference category, the highest odds ratio (OR) among males and females were that of being aged 17–40-year old (OR = 8.76; 95% BCI: 6.03–12.47) and 11–16-year old (OR = 8.59; 95% BCI: 4.74–14.28), respectively. People who did not work on a rice farm had a lower prevalence of infection than those working full time on a rice farm. The OR for irrigated villages compared to rain-fed villages was 1.41 (95% BCI: 0.50–3.21). Discussion We found very important village-to-village variation in prevalence of infection intensity. This variation is probably due to village-level variables other

  5. Study on the genetic diversity of an endangered plant Cercidiphyllum japonicum%濒危植物连香树遗传多样性研究

    Institute of Scientific and Technical Information of China (English)

    黄绍辉; 方炎明

    2011-01-01

    Random amplified polymorphic DNA (RAPD) markers were used to measure the genetic diversity of Cercidiphyllum japonicum, an endangered plant in China. The experiment was implemented among 11 natural populations from Zhejiang, Anhui, Hunan, Hubei, Sichuan, Shannxi and Henan of C. japonicum. A total of 691 DNA fragments were amplified from 20 RAPD primers, of which 328 were polymorphic, account for 47.5 % of all bands. AMOVA was used to determine the variation within and among populations. Results indicated that the variation among populations was 0.479 7and that within populations was 52.03%. The gene flow was 0. 542 4 based on the Nei gene differentiation index indicated that the gene intercourse was difficult among populations. Results of UPGMA cluster analysis indicated that the genetic distance between the population of Shexian Anhui and Jiyuan Henan was the largest among populations. Genetic drift might have enhanced genetic differentiation within populations. In order to protect the species, preserving the habitat and breeding the seedlings to expand the species were suggested.%以我国特有的濒危被子植物连香树为材料,采用RAPD标记对分布于浙江、安徽、湖南、湖北、四川、陕西、河南的11个天然居群进行了检测.以20个引物共扩增出691条DNA片段,其中多态性条带328条,占总条带数的47.5%.AMOVA分析表明,连香树的基因分化系数为0.479 7,居群内变异占总变异的52.03%.由Nei基因分化系数估计的基因流仅为0.542 4,表明居群间基因交流困难.UPGMA聚类分析表明,安徽歙县居群与河南济源居群的关系最远.遗传漂变等因素可能是连香树目前遗传结构的主要成因.保护现有生境、通过培育实生苗并扩大繁殖栽培的范围是目前对连香树比较好的保护措施.

  6. 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

    Directory of Open Access Journals (Sweden)

    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.

  7. Rapid and sensitive assay for the phytotoxin rhizobitoxine.

    OpenAIRE

    Ruan, X.; Peters, N K

    1991-01-01

    Rhizobitoxine is a phytotoxin synthesized by some strains of the legume symbiont genus Bradyrhizobium and the plant pathogen Pseudomonas andropogonis. We demonstrate here a new enzymatic assay which is 100-fold more sensitive than previous assays and can detect as little as 1.0 pmol of rhizobitoxine. The assay is based on the inhibition of Salmonella typhimurium beta-cystathionase by rhizobitoxine. Interestingly, beta-cystathionase from Bradyrhizobium japonicum is insensitive to rhizobitoxine...

  8. Effect of aldicarb on growth and radio-carbon (14C) and radio-phosphorus (32P) assimilation by Rhizobium japonicum

    International Nuclear Information System (INIS)

    In vitro studies on the effect of aldicarb (2-methyl-2-(methyl thio) propionaldehde-o-methyl carbamoyl oxime), a soil applied systemic insecticide, on Rhizobium japonicum revealed that the chemical (at 1,2 and 5 ppm levels) stimulated the growth of the organism initially upto 48 hr which declined thereafter upto 72 hr. The incorporation of 14C-glucose by the cells considerably reduced due to the insecticide treatment. The production of extracellular, water-soluble slime (polysacchardes) was also reduced considerably with increased concentrations of the chemical. However, the incorporation of 14C-radio-activity in the extracellular slime generally enhanced due to the treatment, upto 6 hr after injection of the radioactivity, which declined significantly later at 15 hr, indicating a qualitative difference in the extracellular polysaccharides produced by the insecticide treated cells. The insecticide treatment drastically reduced the incorporation of 32P-disodium hydrogen phosphate into Rhizobium cells, but enhanced the specific activity of the extracellular polysacchrides. (author)

  9. Schistosoma japonicum risk in Jiangsu province, People’s Republic of China: identification of a spatio-temporal risk pattern along the Yangtze River

    Directory of Open Access Journals (Sweden)

    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.

  10. Application of automatic photography in Schistosoma japonicum miracidi-um hatching experiments%自动摄影术在血吸虫毛蚴孵化实验结果观察中的应用

    Institute of Scientific and Technical Information of China (English)

    周明莉; 蔡爱玲; 王雪峰

    2016-01-01

    Objective To explore the value of automatic photography in the observation of results of Schistosoma japoni⁃cum miracidium hatching experiments. Methods Some fresh S. japonicum eggs were added into cow feces,and the samples of feces were divided into a low infested experimental group and a high infested group(40 samples each group). In addition,there was a negative control group with 40 samples of cow feces without S. japonicum eggs. The conventional nylon bag S. japonicum miracidium hatching experiments were performed. The process was observed with the method of flashlight and magnifying glass combined with automatic video(automatic photography method),and,at the same time,with the naked eye observation meth⁃od. The results were compared. Results In the low infested group,the miracidium positive detection rates were 57.5% and 85.0%by the naked eye observation method and automatic photography method,respectively(χ2=11.723,P 0.05). In the two infested groups,the average positive detection rates were 77.5% and 92.5% by the naked eye observation method and automatic photography method,respectively(χ2 = 6.894,P 0.05),80份高低污染组平均阳性检出率分别为77.5%和92.5%(χ2=6.894,P<0.05)。结论自动摄影术可有效提高毛蚴常规尼龙绢集卵孵化实验的阳性检出率。

  11. 水牛和猪感染日本血吸虫后的自愈现象及其机制%The self-cure phenomenon in buffalo and pig infected with Schistosoma japonicum and its mechanism

    Institute of Scientific and Technical Information of China (English)

    吕美云; 李宜锋; 林丹丹

    2010-01-01

    动物感染血吸虫后的自愈现象是指动物在感染血吸虫一段时间后虫负荷数急剧下降、虫体自然清除的现象.水牛和猪作为主要的血吸虫病传染源,在血吸虫病流行传播中起到重要作用.已有研究表明,水牛和猪感染日本血吸虫后有自愈现象,阐明其发生机制对于防治血吸虫病和研制血吸虫疫苗均有重要意义.该文主要围绕水牛、猪感染日本血吸虫后产生的与自愈有关的实验和现场研究作一综述,并对其发生的机制进行探讨.%Self-cure refers to the phenomenon that the number of worms declines sharply and parasites are cleared up naturally after animals infected with Schistosoma japonicum for some time. Buffaloes and pigs play an important role in schistosomiasis transmission as the main sources of infection. Some studies have shown that self-cure occurs in water buffaloes and pigs infected with S. japonicum. It has great significance to clarify the mechanisms behind the phenomenon of self-cure for prevention and control of schistosomiasis and development of anti-schistosomiasis vaccine. This paper summarized the experimental research and field epidemiologi-cal studies around self-cure in water buffaloes and pigs infected with 5. japonicum and discussed the mechanisms of self-cure.

  12. Fluorescent quantitative real-time PCR for quantitative detection of Schistosoma japonicum cercariae in water%荧光实时定量PCR定量水体中日本血吸虫尾蚴方法的建立

    Institute of Scientific and Technical Information of China (English)

    王本敬; 王文波; 周霞; 陈艳勤; 张静; 刘晨晨; 梁幼生; 诸葛洪祥

    2011-01-01

    目的 建立快速、高效、特异的定量水体中尾蚴的数量和检测水体中日本血吸虫尾蚴残余基因组DNA的方法,来评估水体受日本血吸虫尾蚴污染的程度.方法 根据日本血吸虫基因组DNA中的3个多拷贝序列Sjrh1.0(序列号:U92488.1)、18S小亚基单位核楷体核酸基因(18SrRNA)序列(序列号:AY157226.1)和逆转录转座子SjR2的G55A序列(G55A)(序列号:AF412221.1),设计常规PCR引物和实时定量PCR引物,选取较好的靶序列建立SYBR GreenI实时定量PCR方法,绘制尾蚴数的对数与Ct值的标准曲线,并对疫水中日本血吸虫尾蚴残余的基因组DNA进行检测.结果 尾蚴数的对数与Ct值的标准曲线有良好的线性关系,相关系数r2为0.918 6,重复性良好.结论 本方法特异性高,灵敏,可定量水体中尾蚴数,对疫水检测有一定的预警作用.%The objective was to establish a rapid, sensitive and specific method to detect the number of cercariea in water and evaluate level of water stained by Schistosoma japonicum cercariea. Convenience PCR primer sequences were designed tar-geting pSjrH1.0(U92488. 1), Sjl8SrRNA( AY157226. 1) and the clone G55A of the highly repetitive retrotransposon SjR2 (AF412221. 1) in S. japonicum genome, and sequence the PCR product. Based on conserved sequence of pSjrHl. 0, Sjl8SrRNA and clone G55A of highly repetitive retrotransposon SjR2 (G55A), design primers and the quantitative real-time PCR essays were established, by which, the amplifying products were 150 to 170 bp. The sequence in S. japonicum genome and the best annealing temperature were selected by comparing their threshold cycle (Ct value). Then the quantitative real-time PCR essays were established under the better annealing temperature, generate standard curve between the logarithms of gradi-ent diluted DNA templates and Ct value. Five DNA samples extracted from 1, 5, 10, 20 and 80 cercariae were used as quanti-tative template to generate standard curve

  13. Study on the Scavenging Activities of Extracts from Lygodium japonicum(Thunb)Sw. with Different Solvents to Free Radicals%海金沙不同溶剂提取物清除自由基活性的研究

    Institute of Scientific and Technical Information of China (English)

    贲永光; 李康; 李坤平; 张伟永

    2009-01-01

    [目的]比较不同溶剂提取海金沙总黄酮对自由基活性清除能力的大小.[方法]应用DPPH 法、邻苯三酚自氧化法和Fenton反应分别研究了浓度95%乙醇、乙酸乙酯、丙酮、乙酸、氯仿、甲醇提取物清除自由基活性的能力.[结果]6种溶剂提取得到的海金沙提取物对DPPH·、OH·和O2-·均有一定程度的清除作用, 不同溶剂所得提取物对自由基的清除作用均有差别,其中浓度95%乙醇提取得到的海金沙提取物对3种自由基清除效果均最好.[结论]海金沙可作为抗氧化物质资源进行开发利用.%[Objective] The research aimed to compare the scavenging abilities of total flavone from Lygodium japonicum with different solvents to free radical activity.[Method] DPPH method, pyrogallol method and Fenton reaction were used to study the scavenging ability of extracts with 95% ethanol, ethyl acetate, acetone, acetic acid, chloroform, methanol to free radical activity.[Result] Six kinds of solvent extracts from L.japonicum had certain scavenging ability to DPPH·, OH · and O2-·.The scavenging ability of extracted with different solvents to free radicals were difference.The extracts from L.japonicum with 95% ethanol had the best scavenging effects on three kinds of free radicals.[Conclusion] L.japonicum can be used as antioxidants for the development and utilization of resources.

  14. 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

    Directory of Open Access Journals (Sweden)

    Aida T. Rodríguez

    2004-01-01

    Full Text Available 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 soya, a los cuales se les realizaron tres extracciones sucesivas con solventes de polaridad creciente. Se realizó el correspondiente tamizaje fitoquímico y posteriormente se cuantificaron las proteínas y los carbohidratos reductores. Se observó en ambos materiales la presencia de triterpenos, esteroides, quinonas, agrupamientos lactónicos, aminoácidos, aminas, lípidos, aceites, carbohidratos reductores, mucílagos, flavonoides y proteínas. La cuantificación de proteínas y carbohidratos reductores evidenció que el grano molido exhibe mayor concentración de ambos compuestos que la torta desgrasada.

  15. 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.

  16. Generation of a Novel Bacteriophage Library Displaying scFv Antibody Fragments from the Natural Buffalo Host to Identify Antigens from Adult Schistosoma japonicum for Diagnostic Development.

    Directory of Open Access Journals (Sweden)

    Christopher G Hosking

    2015-12-01

    Full Text Available The development of effective diagnostic tools will be essential in the continuing fight to reduce schistosome infection; however, the diagnostic tests available to date are generally laborious and difficult to implement in current parasite control strategies. We generated a series of single-chain antibody Fv domain (scFv phage display libraries from the portal lymph node of field exposed water buffaloes, Bubalus bubalis, 11-12 days post challenge with Schistosoma japonicum cercariae. The selected scFv-phages showed clear enrichment towards adult schistosomes and excretory-secretory (ES proteins by immunofluorescence, ELISA and western blot analysis. The enriched libraries were used to probe a schistosome specific protein microarray resulting in the recognition of a number of proteins, five of which were specific to schistosomes, with RNA expression predominantly in the adult life-stage based on interrogation of schistosome expressed sequence tags (EST. As the libraries were enriched by panning against ES products, these antigens may be excreted or secreted into the host vasculature and hence may make good targets for a diagnostic assay. Further selection of the scFv library against infected mouse sera identified five soluble scFv clones that could selectively recognise soluble whole adult preparations (SWAP relative to an irrelevant protein control (ovalbumin. Furthermore, two of the identified scFv clones also selectively recognised SWAP proteins when spiked into naïve mouse sera. These host B-cell derived scFvs that specifically bind to schistosome protein preparations will be valuable reagents for further development of a cost effective point-of-care diagnostic test.

  17. The development and prospect of the vaccine against Schistosoma japonicum%日本血吸虫疫苗的研究进展与展望

    Institute of Scientific and Technical Information of China (English)

    景英霞; 邵筱

    2010-01-01

    The study on vaccine against Schistosoma has been included into the program of the treat-ment and prevention on the major diseases of WHO and our country with a great progress. The history of the study on vaccines against Schistosoma has passed from dead vaccine, attenuated vaccine, sub-unit vaccine,gene engineered vaccine, nucleic acid vaccine to muhivalent affiliate vaccine. Recently, the research progress on immunologic mechanism of Schiswsoma and the genome of Schistosoma has played a very important role in the research and development of vaccine against Schistosoma. This article reviewed the development of vaccine against Schistosoma japonicum in recent 10 years.%血吸虫疫苗的研究已经纳入了WHO和我国主要疾病防治规划,并且取得了很大的进展.血吸虫疫苗的研究历史经历了从死疫苗、致弱活疫苗、亚单仲疫苗、基因工程疫苗、核酸疫苗到多价联合疫苗等的探索过程.近年来开展的血吸虫免疫机制和血吸虫基因组的研究对血吸虫疫苗的研制起到了积极的推动作用.该文主要对日本血吸虫疫苗近十年的研究进展作一综述.

  18. Cloning of three ZIP/Nramp transporter genes from a Ni hyperaccumulator plant Thlaspi japonicum and their Ni2+-transport abilities.

    Science.gov (United States)

    Mizuno, Takafumi; Usui, Koji; Horie, Kenji; Nosaka, Shiro; Mizuno, Naoharu; Obata, Hitoshi

    2005-08-01

    Ni homeostasis is essential for plant cell activity, but the mechanisms of Ni-transport and delivery are unknown. To elucidate the role of ZIP and NRAMP metal-transporters for Ni2+-transport and homeostasis, we cloned their homologous genes from the Ni hyperaccumulator Thlaspi japonicum, and investigated their Ni-transporting abilities by expression in yeast. The deduced amino acid sequences of the two Zip transporter genes (TjZnt1, TjZnt2) and one Nramp transporter gene cloned had high homologies with TcZNT1 and TcZNT2 of Thlaspi caerulescens and AtNRAMP4 of Arabidopsis thaliana, respectively, and were predicted as integral membrane proteins with 6 or 12 transmembrane domains. TjZNT1 and TjZNT2 had two long histidine-rich domains in the putative cytoplasmic domain between transmembrane domains III and IV. TjNRAMP4 conserved a consensus transporter motif between transmembrane domains VIII and IX. The yeast transformed with TjZNT1 or TjZNT2 showed a marked increase in Ni2+ tolerance with the gene expression. In contrast, the expression of TjNramp4 caused elevation of Ni2+ sensitivity and Ni2+ concentration. These data suggest that ZIP/NRAMP transporters participate in Ni2+ homeostasis of Ni hyperaccumulator plants. TjZNT1 had Zn2+-, Cd2+- and Mn2+-transporting abilities and TjZNT2 also had Zn2+- and Mn2+-transporting abilities, but TjNRAMP4 could transport Ni2+ but not Zn2+, Cd2+ or Mn2+. PMID:16198592

  19. 大豆和花生根瘤菌氢酶的研究%Studies on Hydrogenase from Rhizobium japonicum and Rhizobium arachis

    Institute of Scientific and Technical Information of China (English)

    许良树; 张凤章; 龙敏南; 曾定; 黄河清; 刘月英; 刘广发

    2001-01-01

    根瘤菌在共生固氮过程中因放H2所消耗的能量约占固氮总能量的40%~60%.吸氢酶则能回收和利用固氮过程所放的H2,减少能量损失,从而提高共生固氮效率.在厌氧条件下,加入防止酶蛋白聚合的试剂,利用DEAE-纤维素和SephacrylS-200柱层析,从自养性大豆根瘤菌和花生根瘤菌类菌体中分离并提纯膜结合态氢酶.纯化的两种氢酶表现相近的分子特征:均含有大(60 kD,65kD)、小(30 kD,35 kD)两个亚基;均为NiFe-氢酶,并具有较高的吸H2活性.大豆根瘤菌氢酶的纯酶组分不含Cyt b559.花生根瘤菌L8-3菌株能进行化能自养生长,诱导出高吸H2活性.根瘤菌的吸H2能明显提高固氮活性.从具有高吸H2活性的花生根瘤菌中分离并克隆吸氢基因,采用PCR和探针杂交技术,获得含有吸氢基因的质粒pZ-55.利用多种限制性内切酶构建了质粒pZ-55的物理图谱.通过三亲本杂交,将含吸氢基因的重组质粒转移到不吸H2的花生和毛豆根瘤菌中,所获得的结合株在自生和共生条件下均表达吸H2活性.以结合株接种大田花生,获得的共生根瘤的吸H2活性比接种受体株提高4倍,花生叶片和种子的含N量、产量分别提高1.7%、8.9%和9.6%.%Hydrogen produced by nitrogenase consumed 40~60%0 of energy of symbiotic nitro gen-fixation. Hydrogenase can uptake and reuse the H2 produced by nitrogenase, which results in decreasing the loss of energy and increasing the efficiency of nitrogen fixation. The membranebound hydrogenase from autotrophical cultured Rhizobium japanicum and from the bacteroids of peanut nodule have been purified and characterized. The hydrogenase from R. japonicum consists of two subunits (60 kD, 30 kD). The molecular weight of large and small subunits of hydrogenase from R. arachis is about 65 kD and 35 kD. Both hydrogenases are NiFe-hydrogenase. No cytochrome b(559) could

  20. 1991~2010年江苏省世业镇牛血吸虫感染情况调查%SURVEY ON BOVINE SCHISTOSOMA JAPONICUM INFECTION IN SHIYE TOWN OF JIANGSU PROVINCE FROM 1991 TO 2010

    Institute of Scientific and Technical Information of China (English)

    任雪枫; 高升; 包新奇; 李玉玲; 石锦良; 祝培波; 杨伟华

    2011-01-01

    本文报告了江苏省镇江市丹徒区世业镇1991~2010年牛血吸虫感染情况。世业镇牛血吸虫感染率在不断反复中逐渐降低,到2006年下降至0。在1994~1998年和2002~2005年出现不同程度的反弹,经分析与江水水位变化有密切的关系。观测结果说明江苏省取得了显著血防效果,但血吸虫病防控仍是一项长期任务。%The article reported bovine Schistosoma japonicum infection in Shiye town of Jiangsu Province from 1991 to 2010.The infection rate decreased progressively from 43.36% in 1991 to 0% in 2006.There were two rebounds of infection rate 1994-1998 and 2002-2005,which was largely related to the change of the water level of Changkiang river.The observed results showed that Jiangsu Province had made an outstanding progress in the control of schistosomaiasis,but it had been a long-term task to prevent Schistosoma japonicum infection.

  1. 日本血吸虫和斯氏并殖吸虫的rDNA-ITS2遗传多态性研究%Genetic polymorphism of rDNA-ITS2 of Schistosoma japonicum and Paragonimus skrjabini

    Institute of Scientific and Technical Information of China (English)

    何谐; 张锡林; 牛靖萱; 陈琳

    2009-01-01

    Objective To analyze the genetic variation of rDNA-ITS2 of Schistosoma japonicum and Para-gonimus skrjabini in order to avoid the cross reactions in immune diagnosis between schistosomiasis and paragon-imiasis. Methods The 2 species of fluke were collected, and then the genomic DNA was extracted, to amplify the gene of rDNA-ITS2 by polymerase chain reaction (PCR) and then sequenced. Multiple sequences were aligned by Clusterx, the genetic distance was analyzed by MEGA4, and the phylogentic tree of rDNA-ITS2 genes of Schistosoma and Paragonimus constructed with MP and NJ method. The motifis of the rDNA-ITS2 gene sequences of Schistosoma and Paragonimus were analyzed with DNAsis. Results The results of phylogentic trees of rDNA-ITS2 genes of Schistosoma and Paragonimus were similar. The genetic distance between Schisto-soma japonicum and Paragonimus skrjabini was longer, though there was 42. 7% similarity between their rDNA-ITS2 gene. There were 3 same transcription factors in the sequences of Schistosoma japonicum as well as Para-gonimus skrjabini, AP_2_CS6, bHLH_CS and CAP_site. Conclusion Though there are obviously differences between the Schistosoma japonicum and Paragonimus skrjabini at the level of gene, they have significantly simi-larity in transcription factors of rDNA-ITS2 sequences. It may provide valuable theoretical information for studies on cross-reacting antigen between Schistosoma japonicum and Paragonimus skrjabini at gene level.%目的 分析日本血吸虫和斯氏并殖吸虫的核糖体DNA第二内转录间隔区(rDNA-ITS2)基因的遗传变异.方法 采集2种吸虫成虫标本,提取其基凶组DNA,PCR特异性扩增rDNA-ITS2基因并测序;Clusterx软件进行多序列对比;MEGA 4软件计算遗传距离,NJ法和MP法构建系统进化树;DNAsis软件对rDNA-ITS2序列片段进行模序分析.结果 2种方法构建的遗传系统进化树基本结构相似,日本血吸虫与斯氏并殖吸虫间存在较远的遗传距离,但二吸虫ITS2

  2. 日本血吸虫基因组核糖体移码序列的预测分析%Prediction and analysis of frameshifts in the genome of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    王海彬; 杨忠; 刘锋; 胡薇

    2012-01-01

    目的 预测日本血吸虫基因组中核糖体移码的基因序列并进行鉴定.方法 挑选稳定并能够可靠预测RNA假结结构的软件,编写批量提交数据的程序并结合本地手段进行假结结构预测,计算序列最小自由能从而挑选稳定序列,进一步使用生物信息学软件Fsfinder分析序列中核糖体移码位点,进行开放性阅读框(open reading frame,0RF)分析,筛选出可能产生核糖体移码的日本血吸虫基因序列.利用日本血吸虫蛋白质组数据库中的肽段质谱数据进行比对,寻找对应的肽段信息.结果 从日本血吸虫的8452条基因编码序列中预测出26条可能含有促使核糖体移码假结结构的序列.经过日本血吸虫蛋白质组数据库中的肽段质谱数据进行比对,发现日本血吸虫输入蛋白(Sjimportin)移码之后产生的肽段.结论 整合已有的RNA假结预测软件以及核糖体移码预测软件,建立了日本血吸虫预测核糖体移码序列数据库,并成功获取日本血吸虫蛋白Sjimportin核糖体移码表达证据.%Objective To predict and identify the frameshift sequences in the genome of Schistosoma japonicum.Methods Perl software was used to predict candidate sequences with pseudoknots of S.japonicum automatically.The stable sequences were searched based on the calculation of the minimum free energy and the FSfinder software was used to predict frameshift sites of the sequences,which were compared with proteome database by BLAST.Results From the 8 452 sequences in the S.japonicum database,26 candidate sequences containing both pseudoknot and corresponding frameshift site were selected and the protein Sjimportin bearing frameshift with the evidence in proteome database of S.japonicum was found.Conclusion Combining the RNA pseudoknots prediction software and frameshift prediction software,the database of S.japonicum frameshift has been built and the evidence of frameshift for Sjimportin has been got.

  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. 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.

  5. 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.

  6. 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.

  7. 40 CFR 725.239 - Use of specific microorganisms in activities conducted outside a structure.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Use of specific microorganisms in activities conducted outside a structure. 725.239 Section 725.239 Protection of Environment ENVIRONMENTAL... Bradyrhizobium japonicum. (2) Modification of traits. (i) The introduced genetic material must meet the...

  8. Comparative Study on the Immunogenicity between Recombinant MS-Sj26GST Vaccine and Recombinant BCG-Sj26GST Vaccine in Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    戴五星; 高红; 黄海浪; 袁野; 胡佳杰; 皇甫永穆

    2003-01-01

    The BALB/c mice were immunized with rMS-Sj26GST and rBCG-Sj26GST vaccine inSchistosoma japonicum by subcutaneous injection. After they were immunized for 8 weeks, the eye-balls were removed to get blood and macrophages of abdominal cavity and spleen cells were harves-ted. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating abilityand NO release was used to measure the phagocytic activity of the macrophages. By using ELISAkit, the levels of interleukin-2 (IL-2) and interferon-γ(IFN-γ) in serum and the splenic lymphocyt-ic cultured supernatant were detected. The results showed that after the mice were immunized with106 CFU of rMS-Sj26GST and rBCG-Sj26GST vaccine separately by subcutaneous injection, prolif-erating ability of splenic lymphocytes in the mice showed no difference (P>0.05), but both weresignificantly increased as compared with that in the control group(P<0.05); The contents of NOin the intraperitoneal macrophages of rMS-Sj26GST vaccine group were significantly lower than inthe control group (P<0. 001) and rBCG-Sj26GST vaccine group (P<0. 01); The levels of serumIL-2 in the rMS-Sj26GST vaccine group were significantly increased as compared with that in thecontrol group (P<0. 001), vector group (P<0.01) and rBCG-Sj26GST vaccine group (P<0.05);The contents of serum IFN-γ in the rMS-Sj26GST vaccine group were significantly increased ascompared with that in the control group (P<0.01) and rBCG-Sj26GST vaccine group (P<0.05).The contents of IFN-γ in the cultured supernatant were significantly lower than those of rBCG-Sj26GST vaccine group (P<0. 001), but were significantly increased as compared with that in thecontrol group (P<0.01). It was indicated that both vaccines could enhance the immune response ofthe mice, but rMS-Sj26GST vaccine had stronger immunogenicity than rBCG-Sj26GST vaccine.

  9. 日本血吸虫糖基化磷脂酰肌醇锚定蛋白的鉴定%Identification of Glycosylphosphatidylinositol-anchored Protein from Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    曹勤燕; 薛艳凤; 沈利

    2012-01-01

    目的 鉴定日本血吸虫的糖基化磷脂酰肌醇锚定蛋白(GPI-AP). 方法 根据曼氏血吸虫的糖基化磷脂酰肌醇(GPI)锚定蛋白Sm200的编码基因(GenBank登录号为XM 002569560.1),运用生物信息学方法寻找日本血吸虫的同源基因,分析后选取基因蛋白质编码区(CDS)的部分基因序列(SjGPIs,长约933 bp)进行PCR扩增,并克隆入原核表达载体pET-28a(+),重组质粒转化大肠埃希菌(E.coli) BL21 (DE3)感受态细胞,异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,用镍柱Ni-NTA亲和层析纯化重组肽段SjGPIs.用纯化的重组肽段SjGPIs免疫新西兰大耳兔,以制备的重组肽段抗血清检测日本血吸虫GPI锚定蛋白.用磷脂酰肌醇特异性磷脂酶C(PI-PLC)鉴定检测到的蛋白在日本血吸虫虫体上的锚定方式.检测日本血吸虫感染小鼠的白细胞,确定其是否吞噬GPI锚定蛋白. 结果 日本血吸虫的基因组中存在与曼氏血吸虫GPI锚定蛋白Sm200基因的同源基因序列,经比对拼接后获得3495 bp含完整编码蛋白C末端的基因编码序列.以所选基因序列进行肽段原核表达,获得重组质粒pET-28a(+)-SjGPIs.通过对蛋白C末端序列分析、经蛋白质印迹(Western blotting)分析和PI-PLC酶切验证,发现日本血吸虫被膜存在以GPI形式锚定的蛋白,相对分子质量约为Mr 200000,命名为SjGPI200.感染日本血吸虫小鼠的白细胞中可检测到完整的SjGPI200蛋白. 结论 日本血吸虫存在锚定蛋白SjGPI200,并以GPI形式锚定于虫体被膜上.%Objective To identify glycosylphosphatidylinositol (GPI) anchored protein of Schistosoma japonicum. Methods 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

  10. 扇脉杓兰果实生长动态及胚胎发育过程观察%Observation on fruit growth dynamics and embryo development process of Cypripedium japonicum

    Institute of Scientific and Technical Information of China (English)

    刘芬; 田敏; 王彩霞; 龚茂江; 李全健

    2012-01-01

    对授粉后不同发育阶段扇脉杓兰(Cypripedium japonicum Thunb.)果实的生长动态进行了观察和分析,并分别采用TrC法和常规石蜡切片法研究了种子生活力及其胚胎发育过程.观察结果湿示:扇脉杓兰果实形态成熟时间约为110 d,其中,授粉后0~20 d为第1次迅速生长期,授粉后20~30 d为第1次缓慢生长期,授粉后30~50 d为第2次迅速生长期,授粉后50~110 d为第2次缓慢生长期;果实纵径和横径的生长动态变化过程相似,但横径的生长动态曲线较纵径平缓,形态成熟时果实的纵径和横径分别为48.87和13.59 mm.成熟种子由内外2层种皮和球形胚构成,不具胚乳,内外种皮间具空气腔;败育种子只具有内种皮和外种皮而无种胚.胚胎发育类型为石竹型,种胚自受精形成合子到发育为成熟球形胚约需95 d.种胚发育时合子第1次不均衡横裂形成基细胞和顶细胞;基细胞发育为胚柄细胞,胚柄细胞高度液泡化,在胚胎发育的过程中不进行分裂并逐渐退化消失;顶细胞不参与胚柄形成,并且经过有丝分裂最终形成球形胚;内珠被在种子成熟时发育成为1层致密的紧贴胚体的内种皮.种胚纵径和横径的生长动态变化相似,成熟球形胚的纵径和横径分别为208.71和106.19 μm.扇脉杓兰种子生活力较高,有生活力的种子占56%.根据研究结果推测:自然状态下扇脉杓兰种子萌发率较低,可能与致密的种皮、种子中较小的胚体以及无胚乳导致的营养成分不足有关.%Fruit growth dynamics of Cypripedium japonicum Thunb. At different development stages after pollination were observed and analyzed. And seed viability and embryo development process were also studied by TTC and normal paraffin section methods, respectively. The observation results show that morphological mature time of C. Japonicum fruit is about 110 d, in which, after pollination, 0-20 d is the first fast growth

  11. 日本血吸虫反式剪接前导RNA的鉴定%Identification and Characterization of an mRNA Trans-splicing Leader in Schistosoma Japonicum

    Institute of Scientific and Technical Information of China (English)

    李锐; 贺亮; 郝力力

    2012-01-01

    RNA trans-splicing occurs in a wide range of eukaryotes, from protozoa to chordates. To investigate the SL RNA-mediated trans-splicing in the zoonotic trematode Schistosoma japonicum, a spliced-leader (SL) RNA was cloned in adults using race. The 36-nucleotide SL in S. Japonicum was derived from a 90-nucleotide nonpolyadenylylated RNA transcript encoded by 55 copies of SL genes dispersed in the genome confirmed by blot and real time PCR. Differential transcription patterns of the SL gene in the parasite developmental stages were observed using real time PCR. Predominant expression of SL RNA was found in the two developmental stages, egg and cercariae. Expression of SL RNA transcripts in female was the least. No significant expression difference between male and schistosomulum (3 d and 14 d) was observed. In summary, trans-splicing of spliced leader might acted as an important mechanism of post-transcriptional gene regulation in S. Japonicum.%RNA反式剪接现象广泛存在于真核生物中,包括单细胞原虫以及低等脊索动物.为鉴定日本血吸虫中是否存在SL RNA介导的反式剪接,运用Race方法从成虫中克隆出了1个90 nt的SL RNA基因,36 nt的RNA前导序列正是来源于此90 nt的无PolyA结构的SL RNA,并通过Northern进一步证实了该基因的存在.同时采用荧光定量和Southern对其拷贝数、基因组上的分布方式以及虫体不同阶段的表达量进行了鉴定,发现SL RNA具有55个拷贝并在基因组上呈散在分布;在虫卵和尾蚴时期SL RNA基因的转录丰度最高,雌虫阶段最低,雄虫、3天童虫以及14天童虫阶段无明显差别.结果表明,SL RNA介导的反式剪接可能是日本血吸虫基因转录后重要的调控机制之一.

  12. SEQUENCE ANALYSIS AND PRELIMINARY IDENTIFICATION OF MOLECULES WITH EF-HAND DOMAIN OF SCHISTOSOMA JAPONICUM%日本血吸虫含EF手性结构域分子序列的分析和初步鉴定

    Institute of Scientific and Technical Information of China (English)

    张艳丽; 贾侃; 李莹; 苑纯秀; 杨健美; 林矫矫; 冯新港

    2011-01-01

    为分离鉴定日本血吸虫含EF手性结构域分子及分析其序列结构特征,首先找到含有EF—hand结构域的分子,按照序列一级结构进行分类,之后进行生物信息学分析。然后用PCR方法以虫卵和成虫cDNA文库为模板扩增分类后的分子,构建重组质粒,诱导蛋白表达并纯化,选取14个纯化蛋白用Western blot进行初步免疫原性鉴定。结果269个原始含EF-hand结构域分子按照序列一级结构分为70个;成功表达和纯化了49个含EF手性分子,进化树分析将其分为9类;Western blot显示,14个纯化蛋白均可被日本血吸虫感染小鼠阳性血清识别。本研究结果为下一步采用这类分子进行动物保护性效果评价以及保护性免疫机制的研究奠定了基础。%To isolate and identify the molecules with EF-hand domain of Schistosoma japonicum and analyse the sequence structural characteristics of them, the molecules with EF-hand domain were firstly found and classified according to the primary structure, and then analyzed by bioinformatics. PCR was used to amplify the classified molecules with the S.japonicum adult and egg cDNA library. The recombinant plasmids were constructed and the protein with EF-hand domain were expressed and purified. Western blot was used to identify the preliminary immunogenicity of 14 purified proteins. Two hundred and sixty nine original molecules with EF-hand domain were divided into 70 molecules. Forty nine molecules with EF-hand domain were expressed and divided into 9 categories by phylogenetic tree. Western blot analysis showed that 14 purified proteins were specifically recognized by the positive serum from mice infected with S.japonicum. The study provided the basis for further study of immune protection of the molecules.

  13. 日本血吸虫原肌球蛋白cDNA的克隆及其在大肠杆菌中表达%Cloning of cDNA encoding Schistosoma japonicum tropomyosin and its expression in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    曹建平; 刘述先; 宋光承; 徐馀信

    2002-01-01

    Objective To perform cloning of the gene encod ing Chinese Schistosoma japonicum tropomyosin (SjcTM) and its expression in Escherichia coli.Methods SjcTM cDNA fragment, except for 14 amino acids at the amino terminus, was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) with total RNA extracted from adult worms of S. Japonicum . The RT-PCR product was cloned into T vector and sequenced. The SjcTM cDNA, derived from the constructed TA clone pGEM-SjcTM, was then subcloned into the expressing vector pBV220. After characterization by agarose gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for expression under the temperature-dependent condition. Results The RT-PCR product, cloned into a Tvector, was sequenced and shown to be 96.5% identical at the nuclei acid level and 98.1 % identical in deduced amino acid sequence to that of S. Mansoni tropomyosi n. The target DNA fragment was then subcloned into a prokaryotic vector pBV220 . Induced expression in E. Coli DH5α cells resulted in a constant level of recombinant protein production. The results of SDS-PAGE and Western blot rev ealed that the molecular weight of non-fusion recombinant protein (rSjcTM) was approximately 32 kDa and could be recognized specifically by a polyclonal antise rum specific for native S. Japonicum tropomyosin (SjcTM). Conclusion The engineering of the cDNA encodingS. Japonicum tropomyosin and its bacterial expression was successfully made.%目的克隆日本血吸虫原肌球蛋白编码基因,并在大肠杆菌中表达.方法抽提日本血吸虫(大陆株)成虫总RNA,经逆转录-聚合酶链反应(RT-PCR)获得编码日本血吸虫原肌球蛋白的cDNA片段,该片段与全序列比较,缺氨基端14个氨基酸.该PCR产物克隆入T 载体并对插入片段进行序列测定后,亚克隆入表达载体pbV220,经琼脂糖凝胶电泳、限制性酶切反应和PCR鉴定后,选择克隆用于温控表达.结果 RT-PCR产物

  14. 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

  15. 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.

  16. Phenotype of dendritic cell loaded with antigens from Schistosoma japonicum%日本血吸虫抗原负载树突状细胞的表型研究

    Institute of Scientific and Technical Information of China (English)

    李小红; 曹建平; 汤林华; 周何军

    2010-01-01

    Objective To investigate the phenotype of dendritic cell( DC) loaded with GST and SEA from Schistosoma japonicum. Methods Bone marrow cells were cultured in the presence of IL-4 and GM-CSF to induce dendritic cells(DCs). These DCs were stimulated by purified GST and SEA antigen from Schistosoma japonicum, respectively. FITC labeled anti-GST monoclonal antibody was used to detect the loading of antigen. After that, the expression of CD40, CD80, CD86 and CD11c on the membrane of DC were analyzed by fluorescence activated cell sorting(FACS). Results The loading of antigen was confirmed by the detection of GST on the cells. GST was successfully loaded on DCs. The positive rates of CD40, CD80 and CD86 in the groups stimulated with GST or SEA were not raised significantly, comparing to the PBS control. Conclusion DC loaded with GST and SEA antigen from Schistosoma japonicum exhibited an immature phenotype.%目的 研究负载日本血吸虫谷胱甘肽S-转移酶(glutathione S-transferase,GST)和可溶性虫卵抗原(soluble egg antigen,SEA)的树突状细胞的表型. 方法 骨髓来源的细胞经白介素-4(interleukin-4,IL-4)、粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)诱导培养,获得树突状细胞,体外经GSI、SEA抗原刺激.用异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记的抗GST单克隆抗体染色法检测GST的负载情况.流式细胞仪检测血吸虫抗原负载后树突状细胞表面CD40、CD80、CD86、CD11c分子的表达情况,并与脂多糖(lipopolysaccharide,LPS)、PBS刺激组作比较. 结果 GST负载后在荧光显微镜下可观察到抗GST的特异荧光,表明抗原已被细胞摄取.与LPS相比较,GST、SEA抗原负载后,树突状细胞表面分子CD40、CD86上调不显著,而更类似于PBS刺激组. 结论 日本血吸虫抗原负载后,树突状细胞的表型类似于未成熟表型.

  17. 扇脉杓兰和无距虾脊兰的核型分析%Karyotype Analysis of Cypripedium japonicum Thunb. and Calanthe tsoongiana T. Tang et F. T. Wang

    Institute of Scientific and Technical Information of China (English)

    钱鑫; 连静静; 李全健; 刘芬; 王彩霞; 田敏

    2013-01-01

    为了解扇脉杓兰(Cypripedium japonicum Thunb.)和无距虾脊兰(Calanthe tsoongiana T. Tang et F. T. Wang)的核型,采用根尖压片法对扇脉杓兰和无距虾脊兰的染色体数目和核型进行了研究。结果表明,扇脉杓兰体细胞的染色体数为22,核型公式为2n=2x=22=16m+2sm+2st+2t,染色体相对长度组成为2n=22=2L+6M2+12M1+2S,核不对称系数为60.01%,核型分类为2B型;而无距虾脊兰体细胞的染色体数为40,核型公式为2n=2x=40=28m+10sm+2st,染色体相对长度组成为2n=40=8L+10M2+16M1+6S,核不对称系数为59.84%,核型分类为2B型;两者核型都较为对称。其中,无距虾脊兰的核型为首次报道。这为扇脉杓兰和无距虾脊兰的进化地位和种质保护提供了细胞学证据。%The aim was to understand the karyotype of Cypripedium japonicum Thunb. and Calanthe tsoongiana T. Tang et F. T. Wang. Their chromosome number and karyotype were studied by using ifngertip squashing method. The results showed that the chromosome number of Cypripedium japonium was 2n = 22, karyotype formula was 2n=2x=22=16m+2sm+2st+2t, constitution of relative length was 2n=22=2L+6M2+12M1+2S, asymmetry index was 60.01%, karyotype type was classiifed as 2B;while the chromosome number of Calanthe tsoongiana was 2n=40, karyotype formula was 2n=2x=40=28m+10sm+2st, constitution of relative length was 2n = 40 = 8L+ 10M2+ 16M1+ 6S, asymmetry index was 59.84%, karyotype type also belonged to 2B. Both of them have relatively symmetric karyotype. The karyotype of Calanthe tsoongiana was reported for the first time. These provided some cytological evidences for evolutionary status and germplasm conservation of Cypripedium japonicum and Calanthe tsoongiana.

  18. 日本血吸虫感染鼠巨噬细胞IL-13Rα2的表达%Expression on IL-13Rα2 in macrophages of Schistosoma japonicum-infected mice

    Institute of Scientific and Technical Information of China (English)

    王维; 郑胜生; 祁瑶; 闻惠琴; 刘丽丽; 沈继龙

    2011-01-01

    目的 检测IL-13的诱骗受体IL-13Rα2在血吸虫病巨噬细胞的表达,为从细胞水平探讨Th2型免疫性疾病分子病理机制奠定基础.方法 建立日本血吸虫病鼠模型,采用免疫荧光标记法分别检测肝肉芽肿和原代巨噬细胞IL-13Rα2蛋白的表达情况.结果 荧光显微镜下观察:肝肉芽肿内可见似"咖啡豆样"散在分布的IL-13Rα2免疫阳性信号,巨噬细胞膜内缘呈异常增强的环状荧光.结论 巨噬细胞是IL-13Rα2阳性表达细胞,IL-13Rα2是血吸虫病重要的免疫病理调节分子.%The purpose of this study was to investigate the expression on intracellular proteins of IL-13Rα2 in the macrophages of Schistosomiasis japonicum mice, and to offer the experiment foundation for elucidating IL-13 responding cell and mechanism of fibrogenesis in Th2-mediated diseases. A large number of mice were exposed to cercariae and established S. japonicum mouse model. IL-13Rα2 expressed in the granulomas of liver tissues and peritoneal macrophages was determined by immunofluorescent labeling with primary antibody of goat anti-mouse IL-13Rα2 antibody and monoclonal anti-CD68 antibody,respectively. Coffee bean-like IL-13Rα2 positive staining was showed in the hepatic granuloma in fluorescentmicroscopy. Co-expression of IL-13Rα2 and CD68 protein in peritoneal macrophages from 8-week infected mice was observed. It exhibited the distribution of IL-13Rα2 in cytoplasm concentrated on the inner layer of membrane in macrophages. It's suggested that IL-13Rα2 expressing macrophages might be a critical contributor to the immunopathogenesis of schistosomiasis.

  19. 东亚砂藓茎段长度对繁殖和生长能力影响的测定%Influences of Stem Lengthson on Reproduction and Growth Capacity of Racomitrium japonicum

    Institute of Scientific and Technical Information of China (English)

    夏乔莉; 于天泽; 胡治祥; 于晶; 郭水良

    2014-01-01

    东亚砂藓(Racomitriumjaponicum)是一种适合于立体绿化的观赏藓类植物。通过设置不同强度的切茎处理,形成平均为3.65、5.05和7.77mm的配子体茎段,撒播于表面有薄层泥炭土-蛭石的棕榈垫这一立体绿化载体上,经过85d的培养,比较不同茎段长度下东亚砂藓的生长情况。结果表明,以茎段为5.17mm处理的新枝/旧枝比率、总长度、表面积、枝与叶尖数、鲜重/干重比值最高。因此,今后在对东亚砂藓进行扩繁工作时,茎段长度是一个应重点考虑的因素。%Racomitrium japonicum is a ornamental moss species suitable for vertical greening.Gametophytes were cut into stem sections of 3.65、5.05 and 7.77 mm,respectively,then planted onto vertical planting carri-ers (a palm pad with a thin layer of peat and vermiculite).After a 85-day culture,their growth indices were measured to compare the influences of stem lengths on the growth and reproduction capacity of R.ja-ponicum.Our studies showed that the gametophytes of 5.17 mm in length have highest indices including the length ratio between newly-developed and old stems and branches,total stem and branch length,total area,branch and leaf projection number,fresh weight/dry weight.Therefore,the stem length of gametophytes should be paid attention to in the reproduction of R.japonicum.

  20. 风兰新品种‘红扇’、‘玉金刚’的比较试验研究%Comparative experimental Study on the New varieties of Aerides japonicum' Red fan' and Angraecum Falcatum 'Jade King'

    Institute of Scientific and Technical Information of China (English)

    邱玉宾; 赵庆柱; 林云弟; 韩霞; 赵靖杰; 王炳太

    2012-01-01

    Through the study on comparative experimental of several factors which effect the growth of Aerides japonicum 'Red fan' and Angraecum falcatum 'Jade King', we analyze the diffierence of them. The results showed that it needs to be disinfection treated using potassium permanganate solution before the seedling nutrition planting; Aerides japonieum ' Red fan' is higher than Angraecum falcatum ' Jade King' in survival rate ; The ideal culture medium of two varieties is "moss" and "stone with tree skins", Angraecum falcatum 'Jade King'can also grow well with the cultivation suhstrates of tree skins; The average temperature of culture sites is 11~20 centigrade,h can grow well under condition of 60% shading in summer, two va- rieties have no significant difference; Aerides japonicum ' ease resistance, but there has no significant difference. Red ran'shows batter than Angraecum falcatum 'Jade King' in dis%通过对不同影响因素下红扇、玉金刚风兰的生长比较试验,分析了两风兰新品种的生长差异性。结果表明:定植前用高锰酸钾消毒是保证成活率的有效措施,‘红扇’风兰成活率略高于‘玉金刚’风兰;两品种理想的栽培基质为“苔藓”以及“石块+树皮”,玉金刚采用树皮基质也能获得理想的栽培效果;两品种栽培场所的夜间越冬平均温度在11~20℃的范围内,夏秋季节需遮光60%左右方能正常生长,无差异性;红扇风兰在抗病性方面表现略优于玉金刚风兰,但没有明显的差异。

  1. 青蒿琥酯抗日本血吸虫童虫和成虫作用的研究%Study of the effects of artesunate on juvenile and adult Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    杜海林; 诸葛洪祥; 梁幼生; 魏莉; 周霞; 王廷安

    2010-01-01

    Objective To investigate the details of the therapeutic effects of artesunate against Schistosoma japonicum infection in mice and examine the damage to different stages of S. japonicum. Methods Mice were treated with artesunate 300 mg/kg for 18 d after infection. Twenty-four hours after treatment,schistosomula were collected to determine DNA and protein contents using ultraviolet ray-absorption and Bradford method, respectively. Every 10 schistosomula were maintained in drug-free medium containing 3H thymidine for 24 h ,the tritiated nucleoside uptake and incorporation into nucleic acid were measured using filter membrane and homogenate. Mice infected were orally administered with artesunate at 21 d,the changes of the size of reproduction organ and the development of reproductive cell were investigated. Mice infected with S. japonicum were treated with the same dose of artesunate at 33 d,then the liver tissue sections were examined under a light microscope to investigate the hepatic shift and artesunate-induced alterations in worms. Results One dose of 300 mg/kg artesunate against 18 d schistosomula caused reduction of the DNA and protein contents decreased by 23.0% and 33.6% compared to the control group(P < 0.01). When the above-mentioned schistosomula were in vitro exposed to the tritiated nucleoside for 24 h, apparent decrease in tritiated nucleoside uptake with reduction of 61. 1% (P <0. 01) was seen in the treated schistosomula. The incorporation of 3H thymidine into the schistosomula DNA 24 h after co-incubation was reduced by 40.8% (P <0.01). The significant changes in reproductive system of adult parasites treated at 21 d after infection,suggested that artesunate inhibits sexual maturation. Light microscopic examination revealed that artesunate caused morphological damage to adult ones at 33 d,especially on the worm tegument. Conclusion Artesunate can mitigate the pathogenetic condition and control the spread of schistosomiasis japonicum by

  2. Evaluation of early diagnostic value of 6 antigens from Schistosoma japonicum in mice%血吸虫感染小鼠早期诊断抗原的研究

    Institute of Scientific and Technical Information of China (English)

    王玠; 余传信; 殷旭仁; 钱春艳; 宋丽君; 许永良; 何伟; 曹国群

    2011-01-01

    值,免疫印迹试验的敏感性比酶联免疫吸附试验高.%Objective To find out the candidate antigen for immunoreagent, which could be used to diagnose Schistosoma ja-ponicum infection early in mice. Methods The mice were infected with cereariae of S. japonicum Chinese mainland strain. The sera of mice before and after infection at different time were collected. The recombinant fusion protein (GST-HD) of the large hydro-philic domain (HD) of 23 kDa membrane protein of 5. japonicum with the Glutathione-S-transferase (GST) of S. japonicum, soluble eggs antigen (SEA), TSP2 hydrophilic domain of S. japonkum (TSP2HD), IL4-inducing principle of S. mansoni eggs (IPSE), fusion protein GST-SjMPIO (SjMP-10), and recombinant S. japonicum (Chinese strain) signaling protein 14-3-3 (Sj 14-3-3) were used as diagnostic antigens, the specific IgG and IgM antibodies were measured respectively by enzyme linked immunosorbent assay (ELI-SA). The antigens with the value of diagnosing schistosomiasis early were screened by analyzing the changes of the levels of specific IgG (or IgM) antibodies and the positive rates of specific antibodies in the sera of mice before and post infection at different time. Moreover, the antigen' s value of early diagnosis was further validated by Immunoblot. Results On the 18th, 21st and 28th day post infection, the positive rates of specific antibody IgM against GST-HD were 60%, 70% and 100%, respectively; the positive rates of specific antibody IgG against GST-HD were 40%, 60% and 90%, respectively. The positive rates of antibody IgM againstSEA were 50%, 60% and 90%, respectively; the positive rates of antibody IgG against SEA were 20%, 50% and 70%, respectively. The positive rates of IgM against TSP2HD were 30%, 40% and 50%, respectively; the rates of IgG against TSP2HD were 20%, 30% and 70%, respectively. The positive rates of IgM against IPSE were 20%, 30% and 50%, respectively; the positive rates of IgG against IPSE were 20%, 30% and 60

  3. Protective immunity induced by the anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum%日本血吸虫单克隆抗独特型抗体NP30诱导保护性免疫的研究

    Institute of Scientific and Technical Information of China (English)

    冯振卿; 李玉华; 薛婉芬; 仇镇宁; 李芸茜; 朱昌亮; 管晓虹

    2002-01-01

    Objective To investigate the protective immunity induced by the anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum in mice.Methods An orthogonal table L16(4×212) was selected as the experimental design. Eight-week-old Kunming outbred mice (male and female) were randomly divided into 16 experimental groups and 2 control groups. Control groups were injected with SP2/0 ascites intraperitoneally. Mice from each group were infected with 100±2 cercariae of Schistosoma japonicum in the abdominal skin and were sacrificed on the thirtieth day postchallenge. Adult worms were recovered and counted by perfusion of the left ventricle-portal vein. The SP2/0 ascites injected mice were used as controls and the percentage of protection was calculated.Results Active immunization of mice with NP30 could produce protection levels ranging from 22.36% to 50.46% depending on the different immunity protocols. The best immunization protocol was established from the results.Conclusions Active immunization with NP30 can induce a degree of protection to infection with Schistosoma japonicum cercariae and NP30 is a potential vaccine candidate against Schistosoma japonicum.%目的 观察日本血吸虫单克隆抗独特型抗体NP30主动免疫小鼠诱导的保护性免疫力.方法 选择L16(4×212)正交设计,8周龄昆明种小鼠随机分为16个实验组和2个对照组.对照组腹腔接种SP2/0腹水.所有小鼠在腹部皮肤感染100±2条日本血吸虫尾蚴,尾蚴攻击后第30天处死小鼠,行左心室-门静脉灌注收集成虫,计算减虫率.结果 根据不同的免疫方案,NP30主动免疫可产生22.36%-50.46%的减虫率,并确定了最优免疫方案.结论 提示NP30主动免疫对尾蚴攻击可产生一定的保护力,具有血吸虫疫苗候选分子的潜能.

  4. Effects of Different Media on the Growth of Container Seedlings of Ligustrum japonicum ‘Howardii’%不同基质对金森女贞容器苗生长的影响

    Institute of Scientific and Technical Information of China (English)

    黄军华

    2012-01-01

    以珍珠岩、锯屑、稻壳、粗河砂和泥炭土为材料,采用单形重心混料试验设计方法设计7种轻型基质配方,以常规基质为对照,研究这7种基质对金森女贞容器苗的株高、地径、叶片数、总叶绿素含量等生长指标的影响.结果表明:不同基质配方对容器苗的生长有显著影响,M5(50%稻壳+40%泥炭土+5%珍珠岩+5%粗河砂)、M6(50%锯屑+20%泥炭土+20%稻壳+5%珍珠岩+5%粗河砂)、M7(50%稻壳+20%珍珠岩+20%蛭石+10%粗河砂)混合基质容器苗的生长优于常规基质容器苗,而M1(稻壳)、M2(泥炭土)、M3(锯屑)和M4(东北草炭)单一基质容器苗的生长则显著差于对照.不同基质中金森女贞容器苗的生长分析表明,M5、M6、M7混合基质有适宜的容重、总孔隙度、最大持水量,化学性质稳定,有机质含量较高,为金森女贞容器育苗的优良轻型培养基质,对金森女贞的容器苗生产具有重要意义.%Seven kinds of light media were prepared by using pearlier, sawdust, corn shell, rough sands and peat soil as materials with the simplex centroid mixture design. The growth (height, diameter, leaf number, content of chlorophyll) of the container seedlings was investigated to estimate the effects of 7 media on Ligustrum japonicum 'Howardii', with a traditional medium as the control. The results showed that 7 media significantly influenced the growth of the container seedlings. The growth situations of the seedlings in media M5(50% rice hull + 40 % peaty soil + 5% pearlite + 5% natural river sand) ,M6(50% sawdust + 20% peaty soil+ 20% rice hull+ 5% pearlite + 5% natural river sand) and M7(50% rice hull+ 20% pearlite+ 20% vermiculite + 10% natural river sand) were much better than that in the traditional medium. However,the growth situations in media Ml (rice hull) ,M2( peaty soil) ,M3(sawdust) and M4(dong-bei peat -moss) were worse than that in the control. The analyses of the growth of the

  5. 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.

  6. Construction and Identification of Recombinant Baculovirus with Schistosoma japonicum Sj16 Gene%日本血吸虫Sj16基因重组杆状病毒的构建和鉴定

    Institute of Scientific and Technical Information of China (English)

    胡少敏; 王海; 阮志燕; 余新炳; 吴忠道

    2005-01-01

    目的利用杆状病毒-昆虫细胞系统构建日本血吸虫 (Schistosoma japonicum, Sj)Sj16基因重组杆状 病毒.方法从日本血吸虫尾蚴提取总 RNA,通过 RT- PCR扩增出 Sj16基因全编码区序列,将其克隆到载体 pET30a(+ )中,通过 PCR将 Sj16基因连同 pET30a(+ )多克隆位点下游的 6× His· Tag一起扩增出来,插入 donor载体 pFastBacHTa中,构建重组杆状病毒 donor载体 pFastBacHTa- Sj16- His,转化大肠杆菌 DH10Bac- GFP进行转座,提取重组 Bacmid,用 Lipofectin法转染昆虫细胞 Sf9使其包装成有感染性的重组杆状病毒.结果构建了含日本血吸虫 Sj16基因的重组 Bacmid- GFP- Sj16- His,转染 Sf9细胞后,获得了有感染力的重组杆状病毒. 结论成功构建了日本血吸虫 Sj16基因重组杆状病毒,为下一步重组 Sj16蛋白表达和 Sj16基因功能研究打下了基础.

  7. Growth, nodulation and yield response of soybean to biofertilizers and organic manures

    International Nuclear Information System (INIS)

    A field experiment was conducted to investigate the effect of a symbiotic nitrogen fixing bacterium Bradyrhizobium japonicum strain TAL-102 and a commercial biofertlizer EM (effective microorganisms) on growth, nodulation and yield of soybean [Glycine max (L.) Wilczek] in soils amended either with farmyard manure or Trifolium alexandrinum L. green manure at the rate 20 tons ha/sup -1/ each. In green manure amendment, B. japonicum inoculation significantly enhanced number and biomass of nodules resulting in a significant increase of 27, 65 and 55% in shoot biomass and number and biomass of pods, respectively. In farmyard manure amended soil, B. japonicum inoculation significantly enhanced fresh biomass of nodules. As a result a significant increase of 45 and 47% in shoot biomass and number of pods was recorded, respectively. Generally, the effect of sole EM application on various studied parameters was insignificant in both the soil amendment systems. Combined application of EM and B. japonicum in green manure amended soil reduced shoot growth and number of pods as compared to sole B. japonicum inoculation. Conversely, in farmyard manure amendment, plants co-inoculated with B. japonicum and EM exhibited highest and significantly greater shoot biomass, and number and biomass of pods as compared to all other treatments. The present study concludes that soybean yield can be significantly enhanced by the application of B. japonicum and EM in farmyard manure amendment. (author)

  8. Ageing Down-modulates the Immune Responses to Schistosoma japonicum Infection in Mice%衰老下调小鼠对日本血吸虫感染的免疫应答

    Institute of Scientific and Technical Information of China (English)

    杨云帆; 李雍龙; 孙谦; 苏斌涛; 林琳; 李曼君; 陈琳; 徐宏; 雷家慧; 刘文琪

    2011-01-01

    目的 研究衰老对小鼠感染日本血吸虫后免疫应答的影响.方法 幼龄(2月龄)和老龄(18月龄)雌性BALB/c小鼠各8只,每鼠感染日本血吸虫尾蚴(40±1)条.感染后6周剖杀小鼠,经门静脉灌注收集成虫,计数虫荷;KOH消化法收集肝脏中的虫卵,并计数.制作肝脏连续病理切片,测量两组小鼠肝脏增生期平均单卵肉芽肿的大小,计算单卵肉芽肿体积.常规法制备脾淋巴细胞悬液进行T淋巴细胞增殖实验,计算刺激指数(SI).ELISA法检测脾淋巴细胞中γ干扰素(IFN-γ)和白细胞介素4(IL-4)的表达水平.结果 幼龄组小鼠体内的虫荷和每克肝组织虫卵数分别为26.00±2.42和(2.08±0.87)×104,老龄组虫荷和每克肝脏虫卵数分别为19.75±1.95和(1.59±1.05)×104,两组间的差异有统计学意义(P<0.05).幼龄组肝脏单卵肉芽肿平均体积[(47.02±24.13)×10-3 mm3]明显大于老龄组[(30.13±10.97)×10-3 mm3](P<0.05).T淋巴细胞增殖实验结果 显示,老龄组脾淋巴细胞对ConA的增殖反应(SI:1.08±0.12)低于幼龄组(SI:1.31±0.14),其脾细胞培养上清中IFN-γ和IL-4的含量[(24.05±6.24)、(4.15±0.68) pg/ml]也明显低于幼龄组[(34.25±8.69)、(7.25±0.83) pg/ml] (P<0.05).结论 衰老引起小鼠对日本血吸虫感染的免疫应答下降,从而减轻了日本血吸虫感染引起的免疫病理损害.%Objective To investigate the effect of ageing on the immune responses against Schistosoma japonicum infection in mice. Methods Female BALB/c mice were divided into young group (2 months) and old group (18 months),each composed of 8 mice. Each mouse was percutaneously infected with 40±1 S. japonicum cercanae. At 6 weeks post-infection,the mice were sacrificed, and the spleens were removed and single-cell suspensions of splenocytes were prepared. Worms were perfused from hepatic portal system and counted. The number of eggs in the liver was determined after KOH digestion. Mean single-egg granulomas

  9. IL-6RFP can protect liver function in the model of mice infected with Schistosoma japonicum%IL-6RFP 在小鼠血吸虫感染中对肝细胞的保护作用

    Institute of Scientific and Technical Information of China (English)

    平春光; 程海燕; 高闻达; 计永胜; 刘淼; 任翠平; 邵延靖; 华梦晴; 沈际佳

    2015-01-01

    目的:探讨鼠白介素6受体融合蛋白(mIL-6RFP)在BALB/c 小鼠日本血吸虫感染模型中对肝细胞的保护作用。方法通过亲和层析方法获得可阻断白介素-6(IL-6)的mIL-6RFP,经人类肝癌细胞(HepG2)IL-6刺激及阻断实验检测纤维蛋白原(fibrinogen,FGG)和结合珠蛋白(haptoglobin, HP)的 mRNA 表达变化,体外验证重组蛋白 mIL-6RFP 的生物活性。建立 BALB/c 小鼠日本血吸虫感染模型,于感染第24天起经尾静脉注射 mIL-6RFP,隔天注射每次100μg/只,感染第42天剖杀小鼠。HE 染色法检测小鼠肝脏肉芽肿面积,荧光定量 PCR 法检测小鼠肝组织 IL-1β、IL-13、肿瘤坏死因子-α(TNF-α)和趋化因子1(CXCL1)的 mRNA 表达,连续监测法检测小鼠肝功能指标谷丙转氨酶(ALT)和谷草转氨酶(AST)。结果 HepG2细胞实验显示 mIL-6RFP 可降低IL-6对该细胞的刺激作用,显著下调 FGG 和 HP 的表达(P<0.05)。在小鼠感染模型中阻断 IL-6后,肝功能指标 ALT和 AST 与溶剂对照组相比均显著下降(P <0.05),肉芽肿病变无显著性差异。结论在 BALB/c 小鼠日本血吸虫感染模型中 mIL-6RFP 可明显改善肝细胞功能,但对肉芽肿形成未见明显作用。%Objective To explore the role of mouse interleukin-6 receptor fusion protein (mIL-6RFP)for the pro-tection of liver cells in the model of BALB /c mice infected with Schistosoma japonicum.Methods Nickel ion col-umn was used to obtain purified recombinant mIL-6RFP protein.In vitro,HepG2 cells were used to verify the bio-logical activity of mIL-6RFP.BALB /c mice were administrated by tail vein injection with 100 μg of mIL-6RFP or equal volume solvent,at 24,26,28,30,32,34,36,38,and 40 days after infection.Mice were sacrificed 48 h after the last injection.The granuloma size was measured in the mouse liver by HE staining.Interleukin-1β(IL-1β),interleukin-13 (IL-13 ),tumor

  10. 日本血吸虫尾蚴及童虫可溶性抗原蛋白质组研究%The soluble antigenic proteome of Schistosoma japonicum cercariae and schistosmula

    Institute of Scientific and Technical Information of China (English)

    马安; 王越; 汤益; 杨再峰; 施晓华; 朱明东; 刘晓龙; 干小仙

    2011-01-01

    应用免疫蛋白质组研究方法筛选、鉴定日本血吸虫尾蚴、童虫可溶性抗原蛋白质。方法日本血吸虫尾蚴可溶性粗抗原(SCAP)、童虫可溶性粗抗原(SLAP)分别用双向凝胶电泳(2-DE)分离蛋白质,每样本同时制3块胶,1块胶进行银染,2块胶通过电转印后再分别用感染兔和正常兔血清作Western印迹分析,确定特异性阳性反应点,再从相应银染胶图上找到匹配的抗原蛋白质点;用MALDI-TOF/TOF串联质谱鉴定抗原蛋白质。结果SCAP、SLAP与感染兔血清免疫反应分别获得94个和68个阳性点,在相应的银染胶图上分别获得33个和31个匹配蛋白质点;用MALDI-TOF/TOF串联质谱鉴定和NCBI数据库检索,鉴定成功率分别为100.0%(20/20)和68.2%(15/22)。已获鉴定的SCAP抗原中蛋白酶占62.5%(10/16),SLAP抗原中蛋白酶占36.4%(4/11);2个抗原蛋白为SCAP和SLAP共有。结论2-DE能有效地分离日本血吸虫可溶性抗原蛋白质,2-D Western印迹法能较好地筛选特异性抗原;2-D Western印迹法阳性点与2-DE胶图蛋白质点匹配率低,低丰度的抗原蛋白质易被漏检;尾蚴和童虫的抗原蛋白质组差异较大。%Objective To screen and identify specific antigenic proteins of Schistosoma japonicum cercariae and schistosomula using immunproteomics approaches. Methods Soluble antigenic proteins of Schistosoma japonicun cercarie (SCAP) and 15 days lung-stage schistosomulum (SLAP) were separated by two-dimensional electrophoresis (2-DE). For each sample, three gels were run in parallel with one gel for silver stain and the other two gels for Western blot using Schistosoma japonicum infected rabbit sera and normal rabbit sera separately. The specific antigenic protein spots were determined on the membrane of Western blot. The matched antigenic protein spots on the sliver stained gels were subsequently analysed by MALDI-TOF/TOF-MS/MS respectively. Results 94 and 68 positive

  11. THE SCIENTIFIC BASIS FOR THE CREATION OF NEW FORMS OF MICROBIAL BIOCHEMICALS

    OpenAIRE

    A.P. KOZHEMYAKOV; Yu.V. LAKTIONOV; T.A. POPOVA; A.G. ORLOVA; A.L. KOKORINA; Vaishlya, O. B.; E.V. AGAFONOV; S.A. GUZHVIN; A.A. CHURAKOV; M.T. YAKOVLEVA

    2015-01-01

    Herein we summarize the results of a comprehensive study aimed on the creation of liquid form of biological products for symbiotic and associative rhizobacteria. The objects of study were nodule bacteria from the rhizosphere of galegae (Rhizobium galegae), soybean (Bradyrhizobium japonicum) and associative rhizobacteria (Arthrobacter mysorens, Azospirillum brasilense, Agrobacterium radiobacter), as well as plants of soybean (Glycine max L.), barley (Hordeum L.), alfalfa (Medicago L.), etc. As...

  12. Toxicity of various classes of insecticides to Serangium japonicum,a predator of Bemisia tabaci%不同类型杀虫剂对烟粉虱捕食性天敌日本刀角瓢虫的毒力

    Institute of Scientific and Technical Information of China (English)

    赵建伟; 郑宇; 栗丽娜; 何玉仙; 翁启勇

    2012-01-01

    采用试管药膜法,测定了不同类型杀虫剂对日本刀角瓢虫Serangium japonicum Chapin的毒力.结果表明,灭多威和敌敌畏对日本刀角瓢虫具有极强的毒力,其25%推荐剂量即可导致日本刀角瓢虫全部死亡;氯虫苯甲酰胺、吡虫啉和噻虫嗪对日本刀角瓢虫也具有很强的毒力,其田间推荐剂量可导致日本刀角瓢虫全部死亡,其50%推荐剂量对日本刀角瓢虫的致死率也高达73.33% ~ 100.00%;乐果、烯啶虫胺、丁醚脲田间推荐剂量对日本刀角瓢虫的致死率分别为40.00%、56.67%、50.00%,预示着上述这些杀虫剂的田间应用对刀角瓢虫具有很高的直接杀伤风险.高效氯氰菊酯、联苯菊酯、毒死蜱、甲氰菊酯、高效氯氟氰菊酯、虫螨腈、阿维菌素、吡蚜酮、噻嗪酮、定虫隆和氟虫脲对日本刀角瓢虫的毒力较低甚至没有直接致死作用.研究结果将为合理使用杀虫剂,协调利用化学防治与生物防治对烟粉虱进行综合防治提供依据.%The toxicity of various classes of insecticides to Serangium japonicum Chapin was tested by contact with dry residues on glass tubes. Both methomyl and dichlorovos were highly toxic to S. japonicum with only 25% of the recommended field dose could cause 100% mortality. Chlorantraniliprole, imidacloprid and thiamethoxam caused 100% mortality at the recommended doses and 73. 33% - 100. 00% mortality at 50. 06% of the recommended doses. The recommended doses of dimethoate, nitenpyram, and diafenthiuron caused 40.00% , 56.67% and 50.00% mortality, respectively. Thus, application of any of these insecticides can cause significant mortality in S. japonicum in the field. The other tested insecticides, beta-cypermethrin, bifenthrin, fenpropathrin, beta-cyfluthrin, chlorpyrifos, chlorfenapyr, avermectin, pymetrozine, buprofezin, chlorfluazuron and flufenoxuron, showed relatively low toxicity, or were even not fatal to, S. japonicum. These

  13. Generation and analysis of 113 adult stage Schistosoma japonicum (Chinese strain) expressed sequence tags%113个日本血吸虫(中国大陆株)成虫表达序列标签的获取及分析

    Institute of Scientific and Technical Information of China (English)

    卞国武; 余新炳; 吴忠道

    2002-01-01

    Objective To rapidly and economically obtain knowledge about adult stage Schistosoma japonicum (Chinese strain) expressed genes using expressed sequence tag (EST) .Methods A directional cDNA library constructed from Schis tosoma japonicum (Chinese strain ) adult stage RNA was used to generate expressed sequence tags(ESTs).These were compared against an EMBL-parasites database and GENBANK database by BLASTn and BLASTx. Results A total of 314 phage clones were randomly selected for generating expressed sequence tags(ESTs). From these clones, 132 EST-qu ality sequence were obtained. Among these EST-quality sequences,113 ESTs were successfully submitted to the dbEST at GenBanK. A total of 7.6% of these EST- quality sequences were previously identified sequence of Schistosoma japonicum, while 4.5% were putatively identified sequences of Schistosoma japonicum. A total of 23.5% of these EST-quality sequences were putatively identified sequence of Schistosoma mansoni or other organisms. 57.6% had no matches in the database and were classified as unknown sequences. Most ESTs with the putative prote in identified belonged to housekeeping proteins. Information about several inte resting genes was found. Conclusion Partial cDNA sequencing to generate express ed sequence tags (ESTs) has the potential to rapidly and economically increase our knowledge about adult stage Schistosoma japonicum(Chinese strain) expressed genes.%目的运用表达序列标签(Expressed Sequence Tag, EST)技术快速、节约、有效地获得有关日本血吸虫(中国大陆株)成虫表达基因的知识.方法随机挑取日本血吸虫(中国大陆株)成虫cDNA 文库单个重组克隆进行部分测序以获得EST,获得的EST通过BLAST程序同EMBL寄生虫数据库和GeneBank数据库进行比较及同源性分析.结果本研究共随机挑取日本血吸虫(中国大陆株)成虫cDNA 文库单个重组克隆314个,获得了132个有EST价值的序列,其中113个成功在GeneBank db

  14. Diversity and phylogeny of rhizobia associated with Desmodium spp. in Panxi, Sichuan, China.

    Science.gov (United States)

    Xu, Kai Wei; Zou, Lan; Penttinen, Petri; Zeng, Xiangzhong; Liu, Ming; Zhao, Ke; Chen, Cuiping; Chen, Yuan Xue; Zhang, Xiaoping

    2016-02-01

    Thirty-four rhizobial isolates were obtained from root nodules of four wild Desmodium species growing in Panxi, Sichuan, China. According to the combined ARDRA and IGS-RFLP (CACAI) cluster analysis, Rhizobium, Pararhizobium and Mesorhizobium isolates outnumbered Bradyrhizobium isolates. In general, the isolates representing the same species from the same site clustered together. Furthermore, the four Desmodium species were all nodulated by more than one rhizobial species. AFLP and phenotypic analyses showed that the 34 isolates represented at least 32 distinct strains. None of the strains were found from more than one site or host, indicating a high degree of rhizobial diversity in Panxi. In the multilocus sequence analysis, the isolates were assigned to Pararhizobium giardinii, Bradyrhizobium japonicum, Mesorhizobium septentrionale, and to undescribed species of the genera Rhizobium, Bradyrhizobium and Agrobacterium. PMID:26654528

  15. Amplification of 16S rRNA gene sequences to differentiate two highly related bradyrhizobia species Amplificação de seqüências do gene RNAr 16S para diferenciar duas espécies de bradirrizóbios altamente relacionadas

    Directory of Open Access Journals (Sweden)

    Adriana Giongo

    2007-09-01

    Full Text Available A 16S rRNA gene PCR-based assay was developed aiming at a fast molecular diagnostic method to differentiate the two phylogenetically closely related species Bradyrhizobium japonicum and B. elkanii, isolated from soybean nodules, in order to identify those more competitive and comprising greater nitrogen fixation ability for use in the formulation of commercial inoculants. The assay used was able to discriminate ten reference strains belonging to these two Bradyrhizobium species, as well as to efficiently identify 37 strains isolated from fields cultivated with soybean.Foi avaliado um método baseado em PCR do gene do RNAr 16S, desenvolvido com a finalidade de um diagnóstico molecular rápido na identificação das espécies filogeneticamente relacionadas Bradyrhizobium japonicum e B. elkanii, isoladas de nódulos de soja, visando ao seu emprego na identificação de estirpes mais competitivas e com maior capacidade de fixação de nitrogênio para uso na formulação de inoculantes comerciais. O método usado foi capaz de discriminar dez estirpes-padrão pertencentes a essas duas espécies de Bradyrhizobium, assim como identificar eficientemente 37 estirpes isoladas de lavouras cultivadas com soja.

  16. 大蓟提取物对植物病原真菌的抑菌活性成分研究%Anti-Phytopathogenic Fungi Active Constituent in Extract from Cirsium japonicum DC.

    Institute of Scientific and Technical Information of China (English)

    魏朝霞; 姚茹瑜; 唐嘉义

    2013-01-01

    [目的]大蓟有广泛的生物活性,具有生物农药的开发潜力.确定其中抗植物病原菌的主要活性成分,以期为其在农药领域的开发利用提供依据.[方法]通过溶剂提取法提取,用生长速率法测定提取物对5种植物病原真菌的抑菌活性,并用柱层析法分离活性物质.[结果]在质量浓度50g/L时石油醚提取物对石榴枯萎病菌的抑菌活性最高,抑制率达100%,EC50值为5.1g/L;从中分离出一个三萜类单体化合物,经鉴定为3β,21β-dihydroxyl-20(30)-entaraxastane.[结论]大蓟对植物病原真菌有良好的抑菌活性,所分离到的单体化合物为大蓟中的主要抑菌活性成分.%[Aims] Cirsium japonicum DC.has a wide-spoctrum biological activity and possess exploitation potential to be biopesticide.The research aims to explore the main antifungal component to provide basis for the developent and utilization in pesticide field.[Methods] Solvent extraction method was used to extract active components.A growth rate method was used for determining the antifungal activity of the extraction against 5 kinds of pathogens,and active material was separated by Column Chromatography.[Results] At 50 g/L,petroleum ether extract has the best inhibitory effect on Ceratocystis finbriata with inhibition rate of 100% and with ECh0 value of 5.1 g/L.A triterpene monomer compound was separated and identified as 3β,21β-dihydroxyl-20(30)-en-taraxastane.[Condusions] Daji has an excellent antifungal activity on plant pathogenic fungi,and the monomer compound is the main antifungal component in Daji.

  17. Effect of calcium cyanamid synthetic drug on Schistosoma japonicum egg morphology%氰氨化钙合成药物对血吸虫虫卵形态学影响

    Institute of Scientific and Technical Information of China (English)

    周义生; 彭国华; 胡主花; 冯小武; 朱蓉; 魏望远; 郭家钢

    2015-01-01

    目的:观察虫卵经氰氨化钙合成药物作用后的形态改变,为进一步研究氰氨化钙合成药物杀灭血吸虫虫卵作用机理提供依据。方法向含有血吸虫虫卵的阳性牛粪中加入氰氨化钙合成药物并搅拌,模拟野粪自然形态堆放于洲滩上;同时设空白对照。于1、2、3、7d后分别取样,收集血吸虫虫卵,于显微镜下观察虫卵形态。结果虫卵经氰氨化钙合成药物作用后,颜色逐步加深,毛蚴萎缩,卵壳变厚,3d后胚膜不完整,7d后毛蚴严重变形;对照组毛蚴未见明显萎缩。结论氰氨化钙合成药物对虫卵胚膜及毛蚴有损伤作用,且随时间延长而损伤加重。%Objective To study the morphological change of Schistosoma japonicum eggs processed by calcium cyanamide synthetic drug,so as to provide the basis for further study of the mechanism that calcium cyanamide synthetic drug to schisto⁃some eggs. Methods The calcium cyanamide synthetic drug was added to the cattle feces containing schistosome eggs and mixed up,and then the cattle feces was stacked as original shape on the marshland. Blank controls were set at the same time. The cattle feces samples were collected and the schistosome eggs were observed under a microscope on the 1st,2nd,3rd,7th day after the experiment. Results By the effect of calcium cyanamide synthetic drug,the color of eggs was deepening gradual⁃ly,the miracidia were atrophied,and the shells of eggs were thickened. The embryonic membrane of miracidia was no longer completed 3 days later,and the miracidia were deformed severely 7 days later. The atrophy of miracidia was not obvious in the blank controls. Conclusion The schistosome miracidia and embryonic membrane can be damaged by the calcium cyanamide synthetic drug,and worse damaged with time extending.

  18. Reduction of Total Antioxidant Capacity in Artemether-treated Female Schistosoma japonicum%蒿甲醚诱导日本血吸虫雌虫总抗氧化能力下降

    Institute of Scientific and Technical Information of China (English)

    翟自立; 梅静艳; 焦佩英; 肖树华

    2002-01-01

    目的观察蒿甲醚对日本血吸虫成虫总抗氧化能力的影响.方法体外将血吸虫在含蒿甲醚和氯化血红素的培养液内培养24 h后,或体内感染小鼠经蒿甲醚300 mg/kg治疗6~24 h后,测定虫体的总抗氧化能力.结果体外50μmol/L的蒿甲醚与氯化血红素伍用引起雌虫总抗氧化能力明显下降.体内蒿甲醚作用血吸虫6 h,即见雌虫的总抗氧化能力明显下降.体内、体外试验中,蒿甲醚对雄虫的总抗氧化能力均无影响.结论蒿甲醚诱导雌虫总抗氧化能力下降.%Objective To study the effect of artemether (Art) on total antioxidant capacity (T-AOC) in adult Schistosoma japonicum. Methods In vitro, the T-AOC was determined in five-week old worms incubated without or with Art and/or hemin for 24 h, and the worms were continuously incubated for 96 h, then worm survival was assessed. In vivo, T-AOC was determined in worms freshly recovered from mice 6 - 24 h after treatment with Art 300 mg/kg. Results Throughout 96 h incubation no worms were killed by 50 μmol/L Art or 50 μmol/L hemin alone, but approximatdy 80% of them were killed by Art plus hemin. Addition of reduced glutathione and vitamin E could significantly block the cidal action of the combined treatment. No effect on T-AOC was seen in the worms exposed to Art or heroin alone for 24 h, but the combined treatment led to a pronounced T-AOC reduction in female worms in vitro. Such a drug effect on female worms was demonstrated in vivo. After female worms were exposed to Art for 6 - 24 h in vivo, their T-AOC was significantly reduced by 40% - 64%. However, no drug effect on male worms' T-AOC was observed in vivo and in vitro exposed to Art plus hemin. Conclusion Art-induced T-AOC reduction in female worms may sensitize them to lethal damages of endogenous and exogenous oxygen radicals.

  19. 用关联度和聚类分析法研究连香树人工群落 与环境的关系%Grey correlation and cluster analysis on relationship between Cercidiphyllum japonicum community and its environ mant.

    Institute of Scientific and Technical Information of China (English)

    潘开文; 刘照光

    2001-01-01

    The relationship between 10 years old C. japonicurn plantation forest community and its environmental conditions was studied by grey correlation method and cluster analysis. The reuslts showed that there existed closer relations a mong soil organic matter, available Ca, Fe, Mg and N. Except for N and Mn, all soil available elements tested had no di reet impact on the absorption and accumulation of corresponding elements by the community. The tree species in differ ent habitats was found to absorb soil elements selectively to some extent. The factors affecting the basal diameter, DBH, height and biomass of C. japonicum could be lined as soil chemical characteristics > soil physical properties > cli mate factors, in which, available P had little impact on the growth of C. japonicum. It is suggested that C. japonicum could be restored in the ares with higher air moisture and with fertile porous soil between elevation of 2200 ~ 2700m.%对10年生连香树人工群落与环境关系的研究表明,土壤中有机质、有效Ca、Fe、Ms和N之间有较强 的相关性;土壤有效元素除N和Mn外,一般都不直接影响群落对土壤中各对应的有效元素的吸收和积累.连 香树在不同生境条件下,对土壤元素的吸收有一定的选择性.影响连香树地径、胸径、树高和生物量的因子顺序 表现为土壤化学性质>土壤物理性质>气候因子;土壤有效P对连香树生长的影响小.连香树适宜生长在海拔 2200~2700m,且土壤疏松多孔、富含有机质、自然含水率较高以及空气平均湿度较大的区域.

  20. Analysis of Simple Sequence Repeats in Genomes of Rhizobia

    Institute of Scientific and Technical Information of China (English)

    GAO Ya-mei; HAN Yi-qiang; TANG Hui; SUN Dong-mei; WANG Yan-jie; WANG Wei-dong

    2008-01-01

    Simple sequence repeats (SSRs) or microsatellites, as genetic markers, are ubiquitous in genomes of various organisms. The analysis of SSR in rhizobia genome provides useful information for a variety of applications in population genetics of rhizobia. We analyzed the occurrences, relative abundance, and relative density of SSRs, the most common in Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti genomes se-quenced in the microorganisms tandem repeats database, and SSRs in the three species genomes were compared with each other. The result showed that there were 1 410, 859, and 638 SSRs in B. japonicum, M. loti, and 5. meliloti genomes, respectively. In the genomes of B. japonicum, M. loti, and 5. meliloti, tetranucleotide, pentanucleotide, and hexanucleotide repeats were more abundant and indicated higher mutation rates in these species. The least abundance was mononucleotide repeat. The SSRs type and distribution were similar among these species.

  1. 日本血吸虫SIEA66-68kDa抗原的分离、纯化及免疫保护性研究%Isolation and purification of the 66-68 kDa soluble immature egg antigen Schistosoma japonicum and its protective immunity

    Institute of Scientific and Technical Information of China (English)

    姜孝新; 汪世平; 刘雪琴; 李庆华; 刘明社; 何卓; 彭先楚; 徐绍锐

    2011-01-01

    To investigate the immunoprotection of 66-68 kDa soluble antigen of immature egg of Schistosoma japonicum (SIEA66-68kDa) and to evaluate its feasibility of being used as a molecular vaccine antigen, the SIEA66-68kDa was prepared through SDS-PAGE electrophoresis, protein band resection, electro-elution and ultrafiltration, and the isolated product of protein was used as antigen to immunize Kunming strain of mice. After 3 courses of immunization, mice were challenged with cercariae of S. japonicum, and the reduction rates of worms and eggs were compared with those of the control group with inoculation of PBS. It was found that the mono-specific antibody response against SIEA66-68kDa protein could be induced in mice immunized with this protein. The SIEA66-68kDa could induce significant reduction in adult worms (41.70 %). The rates of egg reduction in the SIEA66-68kDa group reach 51.23%, 59.26% and 45.17% in the liver, intestine tissue and feces, respectively, with significant difference when compared with the control group. It is concluded that the 66-68 kDa immature egg antigen of S. japonicum could protect the Kunming strain of mice from the infection of this parasite, and it could be regarded as the target antigen for the preparation of vaccine against S. japonicum infection.%目的 研究日本血吸虫未成熟虫卵可溶性抗原66-68kDa (SIEA66-68kDa)的动物免疫保护力,并对其作为天然分子疫苗的可行性进行评估.方法 采用电泳切胶、电洗脱、超滤离心等技术,分离纯化SIEA66-68kDa天然分子抗原,免疫小鼠,待其产生抗体后进行尾蚴攻击感染(40尾/只),计算减虫率和减卵率.结果 SIEA66-68kDa天然蛋白质分子能刺激机体产生抗日本血吸虫的作用,其减虫率为41.70%,每克肝脏减卵率为51.23%,雌虫子宫减卵率为29.30%,每克大肠减卵率为59.26%,每克粪便减卵率为45.17%,与对照组相比差异具有统计学意义(P<0.05).结论 SIEA66-68kDa天然分子抗

  2. 3种保虫宿主日本血吸虫特征性差异表达基因的筛选与验证%Screening and verification on characteristic differentially expressed genes of Schistosoma japonicum from three reservoir hosts

    Institute of Scientific and Technical Information of China (English)

    杨健美; 林矫矫; 石耀军; 冯新港; 傅志强; 苑纯秀; 刘金明; 洪炀; 李浩; 陆珂

    2012-01-01

    Objective To get the characteristic differentially expressed genes of Schistosoma japonicum from three important reservoir hosts: yellow cattle, water buffalo and goat, so as to find the genetic markers to identify the various sources of the parasite reservoir hosts. Methods The 49 d worms were collected from artificially infected animals, and the total RNA(s) of worms were extracted and reverse-transcripted to cDNA, and then hybridized with custom-built microarray to screen characteristic differentially expressed genes of every hosl, and the microarray results were validated by the real-time PCR method. Results From results of microarray, we got 3 characteristic differentially expressed genes of S. japonicum from yellow cattle, 4 from water buffalo and 7 from goat. We verified schistosome samples from three reservoir hosts in another experiment, the results showed that 2 in yellow cattle, 3 in water buffalo, and 5 in goat were verified to be consistent with microarray results. Conclusions The ten characteristic differentially expressed genes of S. japonicum from three reservoir hosts screened by microarray might be used as genetic markers to identify the various sources of reservoir hosts for S. japonicum.%目的 筛选日本血吸虫3种重要保虫宿主来源虫体的特征性差异表达基因,寻找可以用于鉴别这3种宿主来源虫体的遗传标记.方法 日本血吸虫尾蚴人工感染黄牛、水牛和山羊,收集感染后49 d成虫,分别抽提RNA,逆转录成cDNA,体外转录为cRNA并进行片段化;采用定制的血吸虫芯片分别进行杂交,筛选每种宿主来源虫体的特征性差异表达基因.采用实时定量PCR法对所筛选差异表达基因的表达水平做进一步测定,验证芯片数据的结果,并对这些特征性差异表达基因进行验证.结果 通过芯片杂交,分析筛选获得实验黄牛组来源虫体特征性差异表达基因3个,水牛组来源虫体4个,山羊组来源虫体7个.其中黄牛组2

  3. 安徽泾县血吸虫病流行区主要传染源调查%Investigation on the infection sources of Schistosomiasis japonicum in Jingxian County, Anhui Province

    Institute of Scientific and Technical Information of China (English)

    夏麦英; 汤国新; 王金苗; 王晓红; 王强; 王玉晔; 胡宗建; 田金海

    2011-01-01

    目的 了解安徽泾县血吸虫病传染源种类,指导血吸虫病防治工作.方法 选择4个野外环境,其中2个有感染性钉螺分布,2个无感染性钉螺分布.调查野粪污染情况,同时调查附近人、家畜及野鼠血吸虫感染情况.人群采用间接血凝试验(IHA)查病;家畜和野鼠采取粪便孵化法进行病原学检查.结果 现场检获牛、狗、兔及不明野生动物阳性野粪,阳性率分别为14.3%、14.8%、8.9%、25.0%;野鼠、牛和人群IHA阳性率分别为,6.9%、15.1%和3.9%.除不明野生动物的野粪外,在有感染螺分布环境的野粪及宿主血吸虫阳性率均高于无感染螺分布的环境.结论 当地血吸虫病流行可能与牛、野鼠、狗、兔、人及其它野生动物有关,各种传染源在传播中的地位有待进一步研究.%Objective To understand the infectious species of Schistosomiasis japonicum in Jingxian county and provide evidence for making out schistosomiasis control strategies. Methods Four environment spots, of which two have infested snails distribution and two has not infested snails distribution, were selected as study pilots. The situation of wild excrement pollution was investigagted. The infection of schistosomiasis of human, domestic cattle and wild mice nearby the environment spots were examined with the method of IHA for human, and hatching technique for domestic cattle and wild mice. Results The positive rate of wild excrements of catde, dog, hare and another unknown wild animal was 14.3%, 14.8%, 8.9%, 25.0%, respectively. The infection rate of wild mice, cattle and human was 6.9%, 15.1%, 3.9%, respectively. The positive rates of wild excrements and host in the sites with infected snails distribution were higher than those in the sites with no infected snails distribution except the excrements of unknown animal. Conclusion It is suggested that cattle, mice, dog, hare, human and other wild animals may contribute to the endemic

  4. Phytocoenological Features of a Rare and Endangered Plant Cinnamomum japonicum var.chinii%珍稀濒危植物普陀樟的群落学特征研究

    Institute of Scientific and Technical Information of China (English)

    黄水文

    2013-01-01

    根据11块样地的野外调查材料,分析了浙江省舟山群岛普陀樟群落的植物区系成分、种类组成、群落结构特征、群落外貌.结果表明:该群落共有维管束植物55种,分属31科48属,其中以热带分布的属占绝对优势,占总属数的52.08%;其生活型中高位芽植物占72.73%,叶级中中型叶与小型叶各占47.27%、45.45%,叶型中单叶占81.82%,复叶占18.18%,叶质中纸质叶与革质叶各占50.91%、41.82%,说明该群落具有显著的热带特征;该群落中灌木层、乔木层、草本层的物种多样性、丰富度、均匀度均偏低,但优势度相对较高;群落垂直结构层次明显,可分为乔木层、灌木层和草本层,但层间植物极度缺乏.%According to the investigation data from 11 typical sampling plots in Zhoushan Islands of Zhejiang Province,the floristie element,the floristie composition,the structure features of community and the physiognomy of Cinnamomum japonicum var.chinii were analyzed.The results showed that the community was composed of 55 vascular plant species belonging to 31 families and 48 genera.The tropic genus was the main genus,which accounts for 52.08% of total genera.In the community,phanerophytes accounts for 72.73% of dominant life form; middle size leaf and small size leaf accounts for 47.27% and 45.45% of dominant leaf size,respectively; single leaf and herbaceous leaf accounts for 81.82% and 18.18% of dominant leaf form,respectively; papery leaf and coriaceous leaf accounts for 50.91% and 41.82%,respectively.Therefore,the community has significant tropic characters.The tree layer,shrub layer and herb layer in the community had relatively lower degree in diversity,abundance and uniformity of species,but had relatively higher degree in dominance.The community vertical structure could be divided into tree layer,shrub layer and herb layer,but interlayer plants extremely lack.

  5. Histological observations on the distribution of three types of haemo-lymphocytes in Oncomelania hupensis (Gastropoda: Pomatiopsidae) infected with Schistosoma japonicum%感染日本血吸虫的湖北钉螺三种血淋巴细胞分布的组织学观察

    Institute of Scientific and Technical Information of China (English)

    Theerakamol Pengsakul; Yassir A Suleiman

    2013-01-01

    Knowledge of defense mechanism of O. hupensis, particularly their haemo-lymphocytes, which are considered to be the main effectors of invertebrate defense system, should be beneficial to control the disease transmission. In this study, qualitative and quantitative investigations of haemo-lymphocytes in negative and positive O. hupensis after infection with S. japonicum was conducted by histological sectioning of infected snails at post-infection days 3, 5, 7, 12, 16, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, and 90. H & E stained haemo-lymphocytes had an appearance with circular nucleus and densely packed granules inside the cell with entire cytoplasm stained deep blue in color. The haemo-lymphocytes were divided into three types based on their nucleus sizes: large, medium, and small, with diameter ranges of 8.1-8.6 μm (large), 5.5-6.0 μm (medium), and 4.2-4.7 μm (small). Post-infection increase in numbers of haemo-lymphocytes in five organs of infected snails including foot, pericardial, gastrointestinal, alimentary gland, and genital organs regions was observed. The rich area of haemo-lymphocytes was in the pericardial region. Moreover, the fewest number of haemo-lymphocytes presented in foot region. The numbers of haemo-lymphocytes with three nucleus sizes in all snail body parts were clearly increased at beginning, then slightly decreased later, and the haemo-lymphocytes with medium nucleus size was the dominated population. The haemo-lymphocytes with large and small nucleus sizes were found but few in number, Data gathered during the present study should be useful in understanding the defense mechanism of O. hupensis in response to S. japonicum infection, and further the research with the aim of controlling the disease more effectively.%目的 湖北钉螺( O.hupensis)的内在防御机制,特别是它们的血淋巴细胞,被认为是无脊椎动物防御系统的主要效应因子,对于控制寄生虫病的传播很有意义.在本次实

  6. 日本血吸虫感染不同相容性动物宿主的比较研究%Comparative study on Schistosoma japonicum infection in different permissive animal hosts

    Institute of Scientific and Technical Information of China (English)

    杨健美; 林矫矫; 苑纯秀; 冯新港; 傅志强; 石耀军; 刘金明; 洪炀; 李浩; 陆珂

    2012-01-01

    The aim was to understand the difference for worm development , worm morphology and host's histopatholo-gy in Schistosoma j aponicum (S. j aponicum) infected animal hosts , including natural hosts and experimental animal hosts . We artificially infected six animal hosts (yellow cattle , water buffalo , goat, New Zealand rabbit , BALB/c mice , and Wistar rat) with the same source of S . japonicum cercariae . The parasites were perfused through the hepatic portal vein on day 49 after infection. The male and female worms were detached manually and counted , and the length and width of the worms were measured . The results showed that the worm recoveries in permissive hosts were higher than that in non permissive hosts , and the length of the worms from permissive hosts were greater than those from non permissive hosts . Parasites in all the hosts could develop into maturation and cause liver damage in their hosts except for Wistar rat . The livers from permissive hosts were fulfilled with white egg nodules , and composed of egg- granulomas ; while the livers from water buffalo were red but with few egg nodules ; the livers from Wistar rat had no damage at all . The histologies! sections from livers of infected natural hosts showed that in yellow cattle and goat , hepatocytes displayed mild swelling , and a large number of inflammatory cells were seen to be in -filtrating and aggregating , including eosinophils and lymphocytes , and typical striped eosinophilic deposits were observed . Compared with yellow cattle and goat , the structural integrity of the hepatic lobules in infected buffalo was intacted in the liv -ers . There was actinomorphous distribution of hepatic cord centered on central veins , polygonal hepatocytes without edema and inflammatory cell infiltration , leaving only scattered neutrophils and monocytes . This study provides reference data for labora- tory and field studies in understanding the differences of S. j aponicum infection in different

  7. Progress in establishment of Microtus fortis experimental model and its applicafion in the rearch of schistosomiasis japonicum%东方田鼠实验动物模型的建立及其在日本血吸虫病研究中的应用

    Institute of Scientific and Technical Information of China (English)

    费正弦; 袁忠英; 胡媛; 曹建平

    2010-01-01

    Microtus fortis is a valuable resource of wild animals in China.To develop an experimental model of Microtus fortis has been listed as a research project in national program and under extensive study.Up to date,schistosomiasis japonicum is an important zoonosis,which remain8 an important public health issue in China.Microtus fortis is a unique wild mammalian resistant to schistosomiasis 80 far.At present,more intensive studies are needed in its taxonomy,anatomy,purification,embryo cryopreservation and transplantation,especiany in molecular genetics to develop a useful experimental model for researches on the resistant mechanism to S.japonicum infection and transgenic breeding of schistosomiasis-resistant animal in laboratory.In this paper.the progress in establishment of Microtus fortis experimental model and its application in the research of schistosomiasis japonica were reviewed.%东方田鼠是我国非常有价值的野生动物资源之一.东方田鼠实验动物模型的建立已被列为国家的有关研究项目并得到了广泛全面的研究.日本血吸虫病是重要的人兽共患寄生虫病,仍是我国一个重要的公共卫生问题.而东方田鼠是迄今为止唯一发现具有天然抗血吸虫病的哺乳动物.目前,在东方田鼠牛物学特性、实验动物化研究、分子遗传学研究方面均有一定的进展,以期为东方田鼠的抗日本血吸虫特性研究提供理想的动物模型.该文就东方田鼠实验动物模型的建立及其在血吸虫病研究中的进展作一综述.

  8. 江苏省血吸虫病监测预警系统的研究Ⅴ长江水域血吸虫毛蚴感染性的监测%Surveillance and forecast system of schistosomiasis in Jiangsu Province V Monitoring of infectivity of Schistosoma japonicum miracidia of Yangtze River

    Institute of Scientific and Technical Information of China (English)

    戴建荣; 洪青标; 梁幼生; 李洪军; 孙乐平; 邢云天; 汪伟; 李幼子; 高扬; 张联恒; 高原

    2011-01-01

    目的 建立长江水体血吸虫毛蚴监测方法,监测水体受血吸虫虫卵污染情况,为追踪与控制传染源、切断传播途径及灭螺提供参考依据.方法 在长江江苏段选择45个监测点,采用浮瓶-尼龙袋哨螺测定法,每个点投放500只阴性钉螺,于2009年5-9月每月投放1次,每次投放28 h,钉螺收回后置于25℃恒温箱内饲养.首次回收2个月后每月采用群体逸蚴法检测1次钉螺感染性,在末次投放3个月后,采用压碎法解剖全部钉螺,观察钉螺血吸虫感染情况,并调查出现感染螺环境的人畜活动情况.建立江苏省长江水域哨螺监测数据库,绘制哨螺阳性点地理分布图,计算江水中钉螺单位时间被血吸虫毛蚴感染的阳性率.结果 45个点5个月共投放哨螺44 717只,回收43 477只,回收率为97.23%;钉螺逸蚴81 410只次,未发现感染性钉螺;解剖钉螺13 033只,发现5只感染血吸虫,钉螺感染率为0.038%,水体中钉螺感染血吸虫的阳性率为4.11/100万.在45个监测点中检出阳性点5个,阳性点出现率为11.11%;其中哨螺阳性点分布在长江南岸3个、北岸1个、江心洲1个,其阳性点出现率分别为21.43%、5.56%和7.69%,南岸哨螺阳性点出现率是北岸的3.8倍.5个阳性点中有3个为渔船民集散地.结论 江苏省长江南岸水域受血吸虫虫卵污染程度高于北岸,渔船民集散地是重要的污染地之一.浮瓶-尼龙螺袋哨螺测定法是监测水体受血吸虫虫卵污染的有效方法.%Objective To establish an approach to monitor the Schistosoma japonicum miracidia in waterbody of the Yangtze River, and monitor the infectivity of water body, so as to provide the evidence for tracing and controlling infection source and interrupting the transmission of schistosomiasis, and Oncomelania snail control. Methods The floating bottle-nylon bag sentinel snails method was employed. A total of 45 surveillance sites were selected along the

  9. Immunoprotection of SjP14 DNA vaccine against Schistosoma japonicum infection in mice%日本血吸虫调宁蛋白样蛋白P14基因疫苗对小鼠免疫保护作用研究

    Institute of Scientific and Technical Information of China (English)

    唐小牛; 汪礼文; 姚勇; 汪学龙

    2012-01-01

    目的 研究日本血吸虫调宁蛋白样蛋白P14基因DNA疫苗对小鼠免疫保护作用.方法 制备无内毒素DNA疫苗,用于免疫小鼠.将雌性BALB/e小鼠随机分为4组,每组10只.生理盐水组给予100μL/鼠/次;空质粒组100 μg/鼠/次、pcDNA3.1 (+)-SjP14组、pcDNA3.1(+ )-SjP14+pcDNA3.1(+)-SjGST组分别给予100μg/鼠/次;每2w免疫一次,共3次.末次免疫后2w.经腹部皮肤感染日本血吸虫尾蚴(30±1)条/鼠.尾蚴攻击6周后解剖小鼠,收集成虫,计算减虫率;同时留取肝脏,部分消化后在显微镜下行虫卵计数,计算减卵率,部分肝脏用于组织病理学分析(HE染色法).观察肝细胞变化及肉芽肿情况.结果 与NS对照组比较,pcDNA3.1(+)-Sj P14p核酸疫苗组减虫率和减卵率分别达到44.6%和61.6%;sjP14核酸疫苗与SjGST核酸疫苗联合免疫组减虫率和减卵率分别提高至56.2%和73.5%.肝组织切片镜下显示,pcD-NA3.1(+)-SjP14组肝脏病变较生理盐水组和空质粒组明显减轻,pcDNA3.1(+)-SjP14+pcDNA3.1(+)-SjGST组肝脏损伤程度最轻,虫卵肉芽肿周围炎症反应轻,肉芽肿面积较小.结论 pcDNA3.1(+)-Sj P14核酸疫苗有一定程度的抗血吸虫感染作用,pcDNA3.1(+)-SjP14与pcDNA3.1 (+)-SjGST疫苗联合免疫能增强小鼠对血吸虫感染的保护.%The aim of this research was to study the immunoprotection of SjVU DNA vaccine against Schistosoma japonicum (S. japonicum) infection in mice. The endotoxin-free DNA vaccines were prepared for mice immunization. The BALB/c mice were randomly divided into 4 groups (n=10), which including saline control group, pcDNA3. 1( + ) group, pcDNA3. K + )-SjP14 group and pcDNA3. K + )-SjP14 + pcDNA3. l( + )-S>GST group. Except for those in control groups, all mice were immunized intramuscularly three times with a 2-week"s intervals. Mice in saline and pcDNA3. 1( + ) groups were given 100 jtg saline and pcDNA3.1(+) per mouse respectively. The pcDNA3. l( + )-5jP14 group was

  10. 3种青蒿素衍生物对日本血吸虫吡喹酮抗性株童虫的体内作用效果观察%In-vivo efficacy of three artesiminin derivatives against schistosomulum of praziquantel-resistant Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    卢萍; 汪伟; 曲国立; 戴建荣; 梁幼生

    2014-01-01

    目的:观察3种青蒿素衍生物双氢青蒿素、青蒿琥酯和蒿甲醚对日本血吸虫吡喹酮抗性株童虫的体内作用效果。方法以经11轮亚治疗剂量吡喹酮筛选的日本血吸虫为吡喹酮抗性株,以未暴露于吡喹酮的日本血吸虫作为吡喹酮敏感株,收集2虫株尾蚴感染小鼠,以300 mg/kg双氢青蒿素、青蒿琥酯和蒿甲醚对感染后7~8 d童虫分别进行2次灌服用药(总剂量600 mg/kg),所有小鼠于感染后45 d解剖,收集小鼠体内成虫并计数,计算减虫率和减雌率。结果300 mg/kg双氢青蒿素、蒿甲醚和青蒿琥酯2日疗法(总剂量600 mg/kg)对日本血吸虫吡喹酮敏感株7~8 d童虫的减虫率为69.8%~71.0%,减雌率为75.4%~79.8%;对日本血吸虫吡喹酮抗性株7~8 d童虫的减虫率为64.6%~66.1%,减雌率为69.3%~71.1%,差异均无统计学意义(均P>0.05)。结论日本血吸虫吡喹酮抗性株对青蒿素类衍生物双氢青蒿素、青蒿琥酯和蒿甲醚依然敏感,青蒿素衍生物与吡喹酮在日本血吸虫中不存在交叉抗药性。%Objective To evaluate the in-vivo efficacy of three artemisinin derivatives artemether, artesunate and dihydroartemisinin against schistosomulum of praziquantel-resistant Schistosoma japonicum,. Methods The S. japonicum subjected to subcurative dose of praziquantel for 11 rounds was served as a praziquantel-resistant strain, while the field-derived strain without exposure to praziquantel was served as a praziquantel-susceptible strain. The mice infected with the two strains of cercaria were treated with artemether, artesunate and dihydroartemisinin at a single dose of 300 mg/kg by gavage in days 7 and 8 post-infection, respectively. All mice were dissected 45 days post-infection, and the adult worms were collected to estimate worm and female worm burden reductions caused by drug treatment. Results Administration of artemether, artesunate and dihydroartemisinin at a single

  11. 日本血吸虫中国株次黄嘌呤鸟嘌呤磷酸核糖转移酶的克隆、表达及其免疫保护性研究%Cloning, Expression and Immunization of The Hypoxanthine-guanine Phosphoribosyltransferase for Schistosoma japonicum Chinese Strain

    Institute of Scientific and Technical Information of China (English)

    余俊龙; 吕志跃; 彭先楚; 周松华; 刘雪琴; 汪世平; 何卓; 戴橄; 李文凯; 姜孝新; 曾少华; 肖小芹; 徐绍锐

    2006-01-01

    A 1 270 bp full-length cDNA fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the '3' and 5' ends of the incomplete expression sequence tag (EST) of hypoxanthine-guanine phosphoribosyltransferase of Schistosoma japonicum (SjHGPRT) were amplified by the anchored PCR with 2 pairs of primer that were designed according to the published incomplete SjHGPRT EST and the sequence of multiclone sites of library λgt1 1 vector. Sequence analysis indicated that this fragment, with an identity of 82% to hypoxanthine-guanine phosphoribosyltransferase ofSchistosoma mansoni (SmHGPRT), contained a complete open reading frame(ORF). The deduced amino acid sequence showed 83% identity to that of SmHGPRT. This fragment was cloned into the prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coli. SDS-PAGE revealed that M of the recombinant protein was about 28 ku. Western-blot analysis showed that the recombinant protein was recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Mice vaccinated with recombinant protein revealed significant worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs of the female worms reduction percentage, compared with the controls. Taken together, the SjHGPRT full-length cDNA can be cloned and expressed in E. coli as a recombinant protein that elicited immunity against the challenge infection with Schistosoma japonicum, indicating its potential as a partia1 protection vaccine candidate.%根据基因库中日本血吸虫次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)EST(BU803192)以及日本血吸虫成虫cDNA文库载体λgt11多克隆位点邻近核苷酸序列设计引物,以日本血吸虫成虫cDNA文库为模板,采用锚式PCR对SjHGPRT基因不完整的3'端和5'端进行扩增、测序,用电子软件拼接,获得SjHGPRT全长cDNA(1 270 bp),经序列分析,推断该片段

  12. Effects of Schistosoma japonicum infection on the expression of Arg-1 and Fizz-1 proteins in liver tis-sues of mice with high-fat diet induced obesity%日本血吸虫感染对高脂饮食小鼠肝组织Arg-1和 Fizz-1表达的影响

    Institute of Scientific and Technical Information of China (English)

    孙科; 赵娜; 王维; 李素梅

    2014-01-01

    Objective To investigate the effects of Schistosoma japonicum infection on selective activation of macrophages and insulin resistance in liver tissues of mice with high-fat diet induced obesity and the possible mechanism. Methods Thirty-six male C57BL/ 6J mice were randomly divided into 3 groups in-cluding normal control group(NC group,n=12),high-fat diet feeding group(HF group,n=12)and high-fat diet feeding with Schistosoma japonicum infection group(HSj group,n=12). Specimens were collected 6 and 12 weeks after high-fat diet feeding. The levels of fasting blood glucose(FBG),fasting plasma insulin (FINS)and homeostasis model assessment for insulin resistance index(HOMA-IR)were detected. The ex-pression of interleukin-6(IL-6),arginase-1(Arg-1)and found in inflammatory zone-1(Fizz-1)at mRNA and protein levels were detected by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry,respectively. Results The mice from HF group showed higher levels of HOMA-IR than those form NC group and HSj group by the end of 6 and 12 weeks after infection(P﹤0. 05). The levels of HOMA-IR in mice from HF group were increased by the end of week 12 after infection as com-pared with those at week 6(P>0. 05). The levels of IL-6 in mice from both HF group and HSj group were higher than those from NC group by the end of week 6 after infection(P﹤0. 05). Higher levels of IL-6 were detected in mice from HF group as compared with those from HSj group and NC group by the end of week 12 after infection(P﹤0. 05). The expression of Arg-1 and Fizz-1 in mice form HF group were lower than those from NC group by the end of 6 and 12 weeks after infection(P﹤0. 05). Arg-1 was highly expressed in mice form HSj group,followed by those from NC and HF groups 12 weeks after infection. The expression of Fizz-1 in mice from HSj group was the highest among the three groups by the end of week 6 and 12 after infection (P﹤0. 05). Conclusion The proinflammatory

  13. 日本血吸虫虫卵、童虫和雌雄成虫膜蛋白的双向电泳%Analysis of membrane proteins from egg, schistosomulum, adult male and female worm of Schistosoma japonicum by two dimensional electrophoresis

    Institute of Scientific and Technical Information of China (English)

    程国锋; 冯新港; 林矫矫; 石耀军; 陆柯; 周元聪; 蔡幼民

    2005-01-01

    Membrane proteins were extracted from eggs, schistosomulum, adult male and female worms of Schistosoma japonicum in order to analyze the differently expressed profile by two dimensional electrophoresis. Schistosomulum and adult worms were obtained from rabbits infected with 1 500 cercariaes on 14 and 42 days after challenge, respectively. Adult male and female worms on 42 days were manually detached and stored into liquid nitrogen until use. Eggs were collected by PercollTM from the liver of rabbits. ProteoPrep Membrane Extraction KitTM was employed to extracted membrane proteins by reducing and alkylating with TBP and iodoacetamide from 200mg of eggs, schistosomulums, adult male worm and female worms, respectively. Immobilized pH gradient strips with a linear pH range of 3-10(130mm) were rehydrated together with membrane proteins (30μg) in 250μl solution containing 7mol urea, 2mol thiourea, 2% SB3-10, 4% CHAPS, 40mmol Tris, 30mmol DTT, then separated on 12.5% SDS polyacrylamide gel for the second dimensional electrophoresis. Gels were stained with silver, scanned by Labscan, and analyzed using ImageMasterTM Analysis software. The 2D maps of egg, schistosomulum, adult female worm and male worm were showed 78±3, 67±3, 108±4 and 122±4 spots respectively. There were 35±1 spots which showed specific expression in female worm as compared with male worm, but 45±2 spots were in male worms. Most differently expressed spots between male and female worms were located in the area of 40-70kD and pI 4-7. The large number of unique spots from sehistosomulum was located in the area of alkalescence. The 2D map of for adult male worms uniquely showed 5 spots as compared with that of schistosomulum and female worm. The female worm showed 4 unique spots as compared with that of schistosomulum, egg and male worm. The unique spots between male and female worms were identified by the database of SWISS 2D-PAGE according to the molecular weight and isoelectronic point

  14. Avaliação da atividade de microrganismos do solo em diferentes sistemas de manejo de soja Evaluation of nitrogen fixation and soil microorganisms in soybean under conventional and minimal cultivation regimes

    OpenAIRE

    O.M. de Castro; H. do Prado; A.C.R. Severo; E. J. B. N. Cardoso

    1993-01-01

    Para avaliar o efeito do manejo do solo e de culturas sobre a atividade de Bradyrhizobium japonicum e de outros microrganismos importantes na ciclagem de nutrientes do solo, montou-se um experimento em casa de vegetação utilizando solo de um ensaio de campo com sistemas de manejo de resíduos e preparo bem distintos, como plantio direto e preparo convencional com arado de discos e diferentes rotações de cultura. O solo é um latossolo roxo distrófico, A moderado, textura argilosa. As culturas u...

  15. STUDIES ON NEW FERTILIZATION AND INOCULATION TECHNIQUES FOR PROMOTION OF GROWTH, SEED YIELD AND QUALITY OF SOYBEAN(Glycine max (L.)Merr.)PLANTS

    OpenAIRE

    Tewari, Kaushal; ティワリ, カウサル

    2006-01-01

    Average soybean seed yield in Japan is less than 2t ha-1, although the seed yield is sometimes much higher up to 6t ha-1 in a well managed field under good climatic conditions. Therefore, the present study was carried out in view to obtain better seed yield by employing the deep placement of different feritilizers such as coated urea, lime nitrogen and urea as well as by using different inoculation methods. For this purpose, ten days old seedling with Bradyrhizobium japonicum inoculated paper...

  16. ProMEX – a mass spectral reference database for Plant Proteomics

    Directory of Open Access Journals (Sweden)

    Stefanie eWienkoop

    2012-06-01

    Full Text Available The ProMEX database is one of the main collection of annotated tryptic peptides in plant proteomics. The main objective of the ProMEX Database is to provide experimental MS/MS-based information for cell type-specific or subcellular proteomes in Arabidopsis thaliana, Medicago truncatula, Chlamydomonas reinhardtii, Lotus japonicus, Lotus corniculatus, Phaseolus vulgaris, Lycopersicon esculentum, Solanum tuberosum, Nicotiana tabacum, Glycine max, Zea mays, Bradyrhizobium japonicum and Sinorhizobium meliloti. Direct links at the protein level to the most relevant databases are present in ProMEX. Furthermore, the spectral sequence information are linked to their respective pathways and can be viewed in pathway maps.

  17. Studies on the effect and mechanism of CD4+ CD25+ regulatory T calls in Schistosoma japonicum immune evasion%CD4+CD25+调节性T细胞在血吸虫免疫逃避中的作用及机制研究

    Institute of Scientific and Technical Information of China (English)

    唐春莲; 郭思洁; 杨进; 祝青; 刘晓宏

    2014-01-01

    目的 探讨CD4+ CD25+调节性T细胞(Tregs)在日本血吸虫免疫逃避中的作用及其机制.方法 雌性BALB/c小鼠随机分成3组,即正常对照组、感染对照组和抗CD25单克隆抗体(anti-CD25 mAb)组,各感染组每只小鼠均经腹部皮肤感染日本血吸虫尾蚴40条,感染后两周anti-CD25 mAb组每只小鼠经腹腔注射anti-CD25 mAb 300 μg,其它组注射等体积的PBS,感染后5周杀鼠冲虫,计数每只小鼠虫荷.收集脾细胞及培养上清,流式细胞术检测脾淋巴细胞中CD4+ CD25+ Tregs百分比.双抗夹心ELISA法测定脾细胞培养上清中的γ-干扰素(IFN-γ)、IL-4、IL-5、IL-10的含量.结果 Anti-CD25mAb组虫荷(23.17 ±6.94)明显低于感染对照组[(30.17 ±5.85),P=0.047];感染对照组脾淋巴细胞中CD4+ CD25+ Tregs百分比(2.68 ±0.12)%明显高于正常对照组[(1.98±0.33%),P=0.049],而anti-CD25mAb组脾淋巴细胞中CD4+ CD25+ Tregs百分比(1.28±0.30)%明显低于感染对照组(P=0.000);anti-CD25mAb组脾细胞培养上清中IFN-γ的含量(386.87±24.85) pg/mL明显高于感染对照组[(61.32±8.75) pg/mL,P=0.000],其余细胞因子组间无统计学意义.结论 anti-CD25 mAb能部分封闭CD4+ CD25+ Tregs后有利于机体清除日本血吸虫,其机制可能为增强Th1型免疫反应,宿主CD4+ CD25+ Tregs有助于日本血吸虫逃避宿主的免疫攻击.%Objective To explore the effect and mechanism of CD4 + CD25 + Tregs in S.japonicum immune evasion.Methods Female BALB/c mice were randomly divided into normal control group,infected control and anti-CD25 mAb group.Each mouse was infected percutaneously with 40 S.japonicum cercaria.After 2 weeks infection,anti-CD25 mAb group was injected intraperitoneal with 300 μg anti-CD25 mAb each mouse.After 5 weeks infection,all mice were succumbed to measure worm burden.The percent of CD4+ CD25 + Tregs in spleen was measured with flow cytometer.The expression of gamma interferon (IFN-γ),interleukin-4 (IL-4),interleukin-5

  18. Scanning electron microscope observation on tegumental damage of 21-d-old Schistosoma japonicum induced by praziquantel%吡喹酮对21-d日本血吸虫童虫皮层损害的扫描电镜观察

    Institute of Scientific and Technical Information of China (English)

    肖树华; 沈炳贵

    1995-01-01

    AIM: To study the effect of praziquantel (Pra) on the tegument of 21-d-old schistosomula, mice infected with Schistosoma japonicum cercariae for 21 d were treated ig with Pra at a single dose. METHOD: Groups of mice were killed at different intervals within 48 h, and the worms were collected by perfusion for scanning electron microscopic observation. RESULTS: When the dose used was moderate swelling, fusion or even erosion and collapsed of the tegumental ridges, which was characterized by swelling of the discoid sensory structures. At higher dose of 500 mg were seen, but more extensive and serious.d, severe swelling, erosion and peeling of the tegument accompanied by the attachment of the host leukocytes on the damaged surface were seen. CONCLUSION: Pra exhibited a direct killing effect on 21-d-old schistosomula.%目的:观察吡喹酮对21-d童虫皮层的作用.方法:小鼠于感染日本血吸虫尾蚴达21 d时,ig1剂吡喹酮,并在治疗后1-48 h的不同时间内剖杀取虫,作扫描电镜观察.结果:吡喹酮的剂量为300 mg·kg-1时,宿主体内的21-d童虫示有轻度或中度的皮层褶嵴肿胀、融合、糜烂或破溃,且以盘状感觉器的肿胀为特征.用吡喹酮的较高剂量500 mg·kg-1治疗,虫的体表亦有相似的变化,但较广泛和严重.若每dig吡喹酮500 mg·kg-1,连给3 d,则虫的皮层严重肿、糜烂和剥落,并伴有宿主的白细胞附着.结论:结果表明,吡喹酮对21-d童虫有直接杀死作用.

  19. 蒿甲醚或吡喹酮早期治疗感染血吸虫尾蚴兔和犬的肝脏显微镜检查%Microscopic observations on livers of rabbits and dogs infected with Schistosoma japonicum cercariae and early treatment with artemether or praziquantel

    Institute of Scientific and Technical Information of China (English)

    肖树华; 杨元清; 张超威; 尤纪青

    1996-01-01

    To study the histopathological change of the liver of the hosts treated with artemether (Art) or praziquantel (Pra) in early stage after infection with Schistosoma japonicum cercariae. METHODS:Dogs infected once with schistosome cercariae were by the repeated dosing once every 1 or 2 wk for 2 -4 times. In rabbits, infected with 48 - 52 schistosome cercariae once every other day for 5 times,infection, followed by the repeated dosing every 1 or 2 wk for 2 - 3 times. RESULTS: After above mentioned dogs or rabbits were treated ig with Art,ArtC or Pra, the female worm reduction rates were 92.1% - 100 %. Histopathological examination showed that the reduction rates of total granuloma in the liver sections of the dogs and rabbits were 70.9 % - 97.3 % and 76.5 % - 97.4 %, respectively. Meantime, the structure of the hepatic lobules was normal with normal arrangement of the liver bundle. CoNCLusIoNS: Early treatment with Art or Pra exhibited a promising effect of protection of the liver of the dogs and rabbits infected with schistosome cercariae.%观察感染血吸虫尾蚴后早期用蒿甲醚(Art)或吡喹酮(Pra)治疗,对宿主肝组织的影响.方法:犬感染198-202条尾蚴后d 7ig Art 10 mg·kg-1,Art胶囊(ArtC)15 mg·kg-1或感染后d 21ig Pra 30-40 mg·kg-1,1-2 wk重复给药1次,共2-4次;兔每隔日感染48-52条尾蚴,共5次,并于第1次感染后d 7或d 21 ig 上述剂量的Art和Pra,停药后4-5 wk剖杀取肝作切片观察.结果:犬与兔经Art或Pra早期治疗后,减♀虫率达92.1%-100%,肝切片中的总虫卵肉芽肿数分别减少70.9%-97.3%和76.5%-97.4%,肝叶结构和肝索排列正常.结论:2种药物早期治疗对宿主肝脏有保护作用.

  20. Investigation on the role of praziquantel against Schistosoma japonicum induced liver fibrosis and expression change of host miRNA%吡喹酮对日本血吸虫病肝纤维化的治疗作用及宿主miRNA表达变化

    Institute of Scientific and Technical Information of China (English)

    陈超; 白瑞璞; 何兴; 潘卫庆

    2015-01-01

    目的 研究吡喹酮对日本血吸虫虫卵引起的肝纤维化的治疗作用及其可能的机制. 方法 选用BALB/c小鼠建立日本血吸虫感染小鼠模型,实验组以250 mg/(kg·d)吡喹酮连续用药3d进行杀虫,再以600 mg/(kg·d)连续给药30 d进行抗纤维化治疗;对照组仅进行吡喹酮杀虫.以肝组织羟脯氨酸含量等指标评定小鼠肝纤维化的程度,并通过实时荧光定量PCR (real-time PCR)检测各组肝组织中某些微小RNA (microRNA,miRNA)的表达水平.各组检测结果以t检验进行统计学分析.结果 经过吡喹酮治疗后,小鼠肝脏羟脯氨酸含量等肝纤维化指标明显降低,小鼠肝脏羟脯氨酸含量:感染后第75天,感染组、杀虫组、治疗组依次为(0.86±0.07)、(0.66±0.06)、(0.25±0.05) mg/g肝湿重,治疗组低于感染组和杀虫组(t=12.86,P<0.01).同时小鼠肝脏的部分miRNA表达水平发生了明显改变,治疗组与杀虫组相比较,Col I、miR-223、miR-146b、miR-142-5p、miR-199a-5p、miR-34c*、miR-195依次下调了62%、38%、75%、77%、40%、54%和56%. 结论 吡喹酮对日本血吸虫病肝纤维化有一定的治疗作用,其作用可能与调节某些宿主miRNA的表达水平有关.%Objective To investigate the role and mechanism of praziquantel against Schistosoma japonicum egg induced liver fibrosis.Methods BALB/c mice were infected with Schistosoma japonicum as experimental mouse model.In the experimental group,praziquantel at 250 mg/(kg·d) for consecutive 3 days as insecticide treatment and at 600 mg/(kg·d) for 30 days of continuous administration as antifibrotic therapy while in the control group,the mice only given praziquantel insecticide treatment.The extent of liver fibrosis was evaluated by detection on liver hydroxyproline content and other indicators,and by detection on certain micro RNA (microRNA,miRNA) expression levels in liver tissues with real-time quantitative PC R (real-time PCR).Test results of each

  1. 血吸虫病肝纤维化小鼠中肝星状细胞迁移功能改变的实验研究%Alterations of hepatic stellate cells on movement abilities in mice infected with Schistosoma ;japonicum

    Institute of Scientific and Technical Information of China (English)

    李兰; 王亚琦; 王洪武; 孙颖; 黄加权; 范翔雪; 宁琴

    2014-01-01

    目的:探讨影响日本血吸虫病肝纤维化小鼠肝组织中肝星状细胞(HSCs)迁移运动功能变化的相关因素。方法 SPF级6~8周龄Balb/c小鼠16只,随机分为模型组(8只)和对照组(8只),以血吸虫尾蚴腹部贴附法建立感染模型,正常组予以生理盐水代替。于感染后8周末处理小鼠,取部分肝组织石蜡包埋,进行病理学评估,免疫荧光染色检测HSCs(α-SMA,红光)运动蛋白Fascin(绿光)的表达;另取部分肝组织,采用Real-time PCR方法检测迁移诱导因子转化生长因子β1(TGF-β1)、血小板源性生长因子(PDGF)以及单核细胞趋化因子1(MCP-1)的表达以及HSCs运动蛋白α-SMA、Fascin的表达。结果8周末时,模型组小鼠肝组织中已形成明显肝纤维化。模型组小鼠肝组织中TGF-β1、PDGF 以及MCP-1的基因表达水平分别是对照组的30倍、14倍及14倍,差异具有统计学意义(P=0.033、P=0.039以及P=0.037);同时,模型组中HSCs运动相关蛋白α-SMA和Fascin的基因表达水平分别是对照组的9倍和5倍,差异具有统计学意义(P=0.004、P=0.018);荧光共聚焦结果提示,模型组小鼠肝组织中α-SMA(红色)和Fascin(绿色)表达部位一致,集中在虫卵周围肝纤维化区域,较对照组二者表达明显增加,且红绿光分布多重叠。结论诱导HSCs运动迁移的因子表达增加和HSCs自身的运动相关蛋白表达增加均有利于HSCs运动迁移能力增强。%Objective To analyze the relevant changes of hepatic stellate cells (HSCs) migration in mice with Schistosoma japonicum infection. Methods A total of 16 SPF balb/c mice aged 6-8 weeks, were randomly divided into two groups, namely, control group (n=8) and infected group (n=8). The mice from the infected group were suffered from skin infection by schistosome cercariae, while the mice in control group were given an equal volume of saline

  2. CONSTRUCTION OF EUKARYOTIC EXPRESSION PLASMID AND SEQUENCE ANALYSIS OF Sj16 GENE OF SCHISTOSOMA JAPONICUM CHINESE STRAIN%日本血吸虫(中国大陆株)Sj16基因真核表达载体的构建及序列分析

    Institute of Scientific and Technical Information of China (English)

    卞国武; 余新炳; 吴忠道; 徐劲; 单志新; 马长玲; 邵筱

    2001-01-01

    目的为了进一步研究日本血吸虫(中国大陆株)Sj16的免疫调节功能,丰富有关血吸虫免疫逃避知识.方法根据曼氏血吸虫Sm1 6基因已知序列设计合成一对引物,用PCR技术从日本血吸虫(中国大陆株)成虫cDNA文库中扩增Sj16基因;将Sj16基因定向克隆入pcDNA3,转化感受态DH5a菌;用酶切、PCR扩增鉴定筛选得到的重组阳性克隆.以重组pcDNA3-Sj16质粒为模板,用双脱氧链末端终止法测定Sj16基因序列,应用软件辅助分析Sj16序列及进行Sj16与曼氏血吸虫的Sm16同源性比较.结果从日本血吸虫(中国大陆株)成虫cDNA文库中获取Sj16基因,重组质粒中含有Sj16基因,成功构建日本血吸虫(中国大陆株)Sj16基因真核表达重组质粒pcDNA3-Sj16;Sj16基因开放读码框有354碱基,编码¨7氨基酸,N端18个氨基酸可能为信号肽序列;Sj16与Sm16有高度同源性,只存在一个氨基酸的差异.结论成功克隆了Sj16基因的真核表达载体,并测定、分析了Sj16基因序列,为深入研究Sj16的免疫调节功能,丰富有关血吸虫免疫逃避知识打下基础.%Objective To further study the immunodulatory function of Schistosoma japonicum Sj16 and more obtain theknowledge about the immune evasion of schistosome. Methods A couple of primers were designed according to theknown sequence of Schistosoma mansoni Sm16 and the Sj16 gene obtained by amplification from cDNA library of S. japon-icum Chinese strain by using PCR technique. By cloning Sj16 gene into a eukaryotic expression vector, pcDNA3, a recombi-nant pcDNA3-Sj16 was constructed and transferred into DH5α. The positive recombinant pcDNA3-Sj16 was screened and i-dentified by agarose gel electrophoresis, endonuclease digestion and PCR technique. The pcDNA3-Sj1 6 recombinant plasmidwas used as template and nucleotide sequence of Sj16 gene was determined by the dideoxy chain termination method. Usingsoftware to analyze the structure and sequence homology of Sj16

  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. Microsymbiont diversity and phylogeny of native bradyrhizobia associated with soybean (Glycine max L. Merr.) nodulation in South African soils.

    Science.gov (United States)

    Naamala, Judith; Jaiswal, Sanjay K; Dakora, Felix D

    2016-07-01

    The genetic diversity and identification of slow- and fast-growing soybean root nodule bacterial isolates from different agro-climatic regions in Mpumalanga, Limpopo and Gauteng Provinces of South Africa were evaluated. The 16S-rDNA-RFLP analysis of 100 rhizobial isolates and eight reference type strains placed the isolates into six major clusters, and revealed their site-dependent genomic diversity. Sequence analysis of single and concatenated housekeeping genes (atpD, glnII and gyrB), as well as the symbiotic gene nifH captured a considerably higher level of genetic diversity and indicated the dominance of Bradyrhizobium diazoefficiens and Bradyrhizobium japonicum in Mpumalanga, Limpopo and Gauteng Provinces. Gene sequence similarities of isolates with type strains of Bradyrhizobium ranged from 97.3 to 100% for the 16S rDNA, and 83.4 to 100% for the housekeeping genes. The glnII gene phylogeny showed discordance with the other genes, suggesting lateral gene transfer or recombination events. Concatenated gene sequence analysis showed that most of the isolates did not align with known type strains and might represent new species from South Africa. This underscores the high genetic variability associated with soybean Bradyrhizobium in South African soils, and the presence of an important reservoir of novel soybean-nodulating bradyrhizobia in the country. In this study, the grouping of isolates was influenced by site origin, with Group I isolates originating from Limpopo Province and Groups II and III from Mpumlanga Province in the 16S rDNA-RFLP analysis. PMID:27324571

  5. A study of the effects of Schistosoma japonicum soluble egg antigen on CD4+CD25+regulatory T cells in patients with asthma%CD4+CD25+调节性T细胞在血吸虫可溶性虫卵抗原影响哮喘中的作用研究

    Institute of Scientific and Technical Information of China (English)

    朱云娟; 刘佩梅; 杨秀珍; 刘霞; 吴增强; 纪伟华; 安桂珍; 沈悦云; 刘金霞; 李健

    2011-01-01

    Objective To study the relationship between Schistosoma japonicum soluble egg antigen (SEA) and asthma and the effects of SEA on CD4+ CD25+ regulatory T cells (CD4+ CD25+ Treg) and expression of the Foxp3 gene. Methods BALB/C mice were each injected with 50 μg SEA peritoneally and through the foot pad once a week for 4 weeks. In the control group, all injections were with normal saline. Then asthma was induced with ovalbumin (OVA) in all mice. After mice were sacrificed, the lungs were subjected to pathologic examination; the bronchoalveolar lavage fluid (BALF) was collected and different cells were classified and counted after smearing and staining. Spleen cells were separated and the percentage of CD4+ CD25+ Treg out of total CD4+ T cells was determined using flow cytometry. Total spleen RNA was prepared and synthesized into cDNA through reverse transcription; cDNA was subjected to PCR amplification to determine the level of Foxp3 mRNA expression. Results Mild pulmonary inflammation was observed in the SEA immunization group, whereas severe inflammation was observed in the control group. Staining of the BALF revealed that the SEA immunization group had a much lower BALF cell density than did the control group. In the SEA immunization group, the percentage of eosinophils out of total cells was(2. 22± 1. 52)% while it was (19. 93±4. 08)% in the control group. The difference between the 2 groups was statistically significant (P<0. 05). Flow cytometry revealed that the percentage of CD4+ CD25+ Treg out of total CD4+ T cells was (32. 24±2. 19) % in the SEA immunization group while it was (27. 41±2. 87) % in the control group. The difference between the 2 groups was statistically significant (P<0. 05). The level of Foxp3 mRNA expression was higher than that in the control group as well. Conclusion SEA inhibits the development of asthma to some extent and it seems influence immune regulation through its effect on CD4+CD25+Treg.%目的 研究血吸虫可溶性虫

  6. Protective effect of SjC23-Hsp70 DNA vaccine and interleukin-12 on Schistosoma japonicum infection in water buffalos%日本血吸虫SjC23-Hsp70 DNA疫苗与IL-12对水牛保护性作用的研究

    Institute of Scientific and Technical Information of China (English)

    胡平成; 夏大; 崔虹艳; 张苹芳; 何永康; 喻鑫玲; 孙振球

    2012-01-01

    目的:研究日本血吸虫中国大陆株23 kD膜蛋白-热休克蛋白(SjC23-Hsp70)DNA疫苗联合佐剂白细胞介素12(IL-12)质粒DNA对水牛的免疫保护作用.方法:将血吸虫病非流行区8~10月龄健康水牛45头随机分为A组(SjC23-Hsp70+IL-12)、B组(SjC23+IL- 12)和C组(pVAX+IL-12),每组15头.每头牛经肩部肌注免疫3次,每次间隔28 d.末次免疫后28 d,每头牛感染日本血吸虫尾蚴1000条.解剖前2天及当天分别收集粪便1次,用定量法检测虫卵和毛蚴数.攻击感染后56天解剖所有水牛,经胸主动脉灌冲法收集成虫,计数成虫数,检测每克肝组织虫卵数.结果:A,B组与C组相比,分别获得45.70%和26.61%的减雌率,44.51%和25.84%的减虫率,41.10%和31.63%的减粪卵率,48.11%和38.07%的减毛蚴率及43.39%和31.95%的减肝卵率.A组的5个率均比B组高(P<0.05).结论:用SjC23-Hsp70 DNA疫苗和IL-12联合免疫水牛可获得明显的免疫保护作用.%Objective: To determine the immune-protective effect of Japan Schistosoma (Chinese mainland strain) 23 kD membrane protein-heat shock protein (SjC23-Hsp70) DNA vaccine plus adjuvant-induced interleukin-12 (IL-12) plasmid DNA on Schistosoma japonicum infection in water buffalos. Methods: Forty-five health water buffalos (8-10 months old) in non-endemic area of schistosomiasis were randomly assigned into group A (SjC23-Hsp70+IL-12, 300 μg), group B (SjC23+IL-12, 300 μg) and group C (pVAX+IL-12, 300 μg), 15 in each group. Each buffalo was immuned by shoulder intramuscular injection for 3 times, at an interval of 28 days. Twenty-eight days after the last immunization, each buffalo was infected with 1000 Japan cercariae of Schistosoma. Fecal examinations were conducted 2 days and 1 day before the perfusion, and on the day of perfusion. The number of hatching miracidia and eggs per gram feces was recorded. Fifty-six days after the infection, the buffalos were sacrificed and perfused via the descending

  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-U14747-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 9e-08 CP000614_1743( CP000614 |pid:none) Burkholderia vietnamiensis G4 c... 60 9e-08 (Q9NYR8) RecName: Full=Retinol...enase/reductase SDR family member... 56 2e-06 CP000615_1850( CP000615 |pid:none) Burkholderia vietnamiensis ...16 |pid:none) Thermotoga neapolitana DSM 4359,... 57 1e-06 CU640366_578( CU640366 |pid:none) Podospora anserina genomic...40 |pid:none) Bradyrhizobium japonicum USDA 1... 56 2e-06 BA000030_774( BA000030 |pid:none) Streptomyces avermitili... 2 ( AC006909 ) Caenorhabditis elegans clone Y82E9, *** SEQUENCIN... 38 1.5 8 ( AC215467 ) Solanum lycopersic

  9. Dicty_cDB: Contig-U14902-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 0 CP000614_1201( CP000614 |pid:none) Burkholderia vietnamiensis G4 c... 38 0.30 CP001400_215( CP001400 |pid:none) Sulfol...ma heat shock pro... 50 6e-05 CP000916_305( CP000916 |pid:none) Thermotoga neapolitana DSM 4359,....7961( BA000040 |pid:none) Bradyrhizobium japonicum USDA 1... 50 6e-05 AF106330_2( AF106330 |pid:none) Thermotoga mariti...512 |pid:none) Thermotoga maritima MSB8, comple... 50 8e-05 AP008934_503( AP008934 |pid:none) Staphylococcus saprophytic...furreducens PCA, co... 43 0.009 AE017226_1020( AE017226 |pid:none) Treponema denticol

  10. 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.

  11. PHENOTYPIC AND BIOCHEMICAL CHARACTERIZATION OF BRADYRHIZOBIUM AND ENSIFER SPP. ISOLATED FROM SOYBEAN RHIZOSPHERE

    Directory of Open Access Journals (Sweden)

    Harpreet Kaur

    2012-01-01

    Full Text Available A total of 15 rhizobia were isolated from soybean rhizosphere on yeast extract mannitol agar (YEMA medium. The isolates were further subjected to morphological, cultural and biochemical characterization along with two reference culture (DS-1, National check and SB271(Local check. Out of 15 isolates, 10 were selected as rhizobia on ketolactose medium (circular, light pink colonies, further 6 (LSER4,LSER5,LSER6,LSER7,LSER8,LSER9 were selected as fast and 4 (LSBR1, LSBR2, LSBR3, LSBR10 as slow growing isolates based on bromothymol blue (BTB test. Fast and slow growing rhizobia were found to be positive for oxidase, urease, citrate utilization and catalase whereas negative reactions for methyl red and voges-proskauer test. The optimum physical condition for growth of fast and slow growing rhizobia was 28˚C at neutral pH (7.0. Four bradyrhizobial spp. and one Ensifer spp. showed hydrogen uptake (Hup+ positive system with 0.01% TTC dye.

  12. Nylig isolerte stammer av Bradyrhizobium viser stor diversitet i fenotypiske karakterer ved denitrifikasjon

    OpenAIRE

    Heen, Guro

    2012-01-01

    Sammendrag Denitrifikasjon er en respiratorisk prosess som utføres av mange prokaryoter under anoksiske forhold. Under denitrifikasjonsprosessen reduseres nitrat (NO3 - ) til nitritt (NO2 - ) og videre til gassene nitrogenmonooksid (NO), lystgass (N2O) og molekylært nitrogen (N2). For å utføre en fullstendig denitrifikasjon kreves de fire reduksjonsenzymene: nitrat reduktase (Nar/Nap), nitritt reduktase (Nir), nitrogenmonooksid reduktase (Nor) og dinitrogenoksid...

  13. Cloning of nod gene regions from mesquite rhizobia and bradyrhizobia and nucleotide sequence of the nodD gene from mesquite rhizobia.

    Science.gov (United States)

    Thomas, P M; Golly, K F; Virginia, R A; Zyskind, J W

    1995-09-01

    Nitrogen-fixing symbiosis between bacteria and the tree legume mesquite (Prosopis glandulosa) is important for the maintenance of many desert ecosystems. Genes essential for nodulation and for extending the host range to mesquite were isolated from cosmid libraries of Rhizobium (mesquite) sp. strain HW17b and Bradyrhizobium (mesquite) sp. strain HW10h and were shown to be closely linked. All of the cosmid clones of rhizobia that extended the host range of Rhizobium (Parasponia) sp. strain NGR234CS to mesquite also supported nodulation of a Sym- mesquite strain. The cosmid clones of bradyrhizobia that extended the host range of Rhizobium (Parasponia) sp. strain NGR234CS to mesquite were only able to confer nodulation ability in the Sym- mesquite strain if they also contained a nodD-hybridizing region. Subclones containing just the nodD genes of either genus did not extend the host range of Rhizobium (Parasponia) sp. to mesquite, indicating that the nodD gene is insufficient for mesquite nodulation. The nodD gene region is conserved among mesquite-nodulating rhizobia regardless of the soil depth from which they were collected, indicating descent from a common ancestor. In a tree of distance relationships, the NodD amino acid sequence from mesquite rhizobia clusters with homologs from symbionts that can infect both herbaceous and tree legumes, including Rhizobium tropici, Rhizobium leguminosarum bv; phaseoli, Rhizobium loti, and Bradyrhizobium japonicum. PMID:7574650

  14. EFECTO DE DIFERENTES INOCULANTES SOBRE LA NODULACIÓN DE LA SOYA CULTIVADA EN CONDICIONES DE ESTRÉS

    Directory of Open Access Journals (Sweden)

    María C. Nápoles García

    2014-01-01

    Full Text Available La soya es un cultivo que se asocia naturalmente con bacterias del género Bradyrhizobium , a través de una simbiosis en la que la planta garantiza fuente de nutrientes y energía al microorganismo y este le aporta nitrógeno que fija de la atmósfera. Además de múltiples señales moleculares que rigen la interacción entre la planta y el microorganismo, las condiciones del suelo influyen sobre el éxito o el fracaso en la nodulación. En este trabajo se estudió el efecto de diferentes inoculantes a base de Bradyrhizobium japonicum sobre la nodulación de la soya, variedad Pioneer 94M30, ante condiciones adversas como bajas temperaturas, acidez y exceso de agua. Se observó que el empleo de inoculantes inducidos influyó positivamente sobre los parámetros de nodulación evaluados en las condiciones de estrés impuestas, así como que el efecto de diferentes dosis del inoculante depende del tipo de estrés y de su intensidad. Estos resultados, aunque preliminares, mostraron un posible efecto antiestrés de los inoculantes inducidos ante tales condiciones ambientales.

  15. Cloning of nod gene regions from mesquite rhizobia and bradyrhizobia and nucleotide sequence of the nodD gene from mesquite rhizobia.

    Science.gov (United States)

    Thomas, P M; Golly, K F; Virginia, R A; Zyskind, J W

    1995-09-01

    Nitrogen-fixing symbiosis between bacteria and the tree legume mesquite (Prosopis glandulosa) is important for the maintenance of many desert ecosystems. Genes essential for nodulation and for extending the host range to mesquite were isolated from cosmid libraries of Rhizobium (mesquite) sp. strain HW17b and Bradyrhizobium (mesquite) sp. strain HW10h and were shown to be closely linked. All of the cosmid clones of rhizobia that extended the host range of Rhizobium (Parasponia) sp. strain NGR234CS to mesquite also supported nodulation of a Sym- mesquite strain. The cosmid clones of bradyrhizobia that extended the host range of Rhizobium (Parasponia) sp. strain NGR234CS to mesquite were only able to confer nodulation ability in the Sym- mesquite strain if they also contained a nodD-hybridizing region. Subclones containing just the nodD genes of either genus did not extend the host range of Rhizobium (Parasponia) sp. to mesquite, indicating that the nodD gene is insufficient for mesquite nodulation. The nodD gene region is conserved among mesquite-nodulating rhizobia regardless of the soil depth from which they were collected, indicating descent from a common ancestor. In a tree of distance relationships, the NodD amino acid sequence from mesquite rhizobia clusters with homologs from symbionts that can infect both herbaceous and tree legumes, including Rhizobium tropici, Rhizobium leguminosarum bv; phaseoli, Rhizobium loti, and Bradyrhizobium japonicum.

  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. 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.

  18. Fe-phyllosilicate redox cycling organisms from a redox transition zone in Hanford 300 Area sediments

    Energy Technology Data Exchange (ETDEWEB)

    Benzine, Jason; Shelobolina, Evgenya S.; Xiong, Mai Yia; Kennedy, David W.; McKinley, James P.; Lin, Xueju; Roden, Eric E.

    2013-01-01

    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. 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.

  20. A bioinformatics insight to rhizobial globins: gene identification and mapping, polypeptide sequence and phenetic analysis, and protein modeling. [v1; ref status: indexed, http://f1000r.es/5ai

    Directory of Open Access Journals (Sweden)

    Reinier Gesto-Borroto

    2015-05-01

    Full Text Available Globins (Glbs are proteins widely distributed in organisms. Three evolutionary families have been identified in Glbs: the M, S and T Glb families. The M Glbs include flavohemoglobins (fHbs and single-domain Glbs (SDgbs; the S Glbs include globin-coupled sensors (GCSs, protoglobins and sensor single domain globins, and the T Glbs include truncated Glbs (tHbs. Structurally, the M and S Glbs exhibit 3/3-folding whereas the T Glbs exhibit 2/2-folding. Glbs are widespread in bacteria, including several rhizobial genomes. However, only few rhizobial Glbs have been characterized. Hence, we characterized Glbs from 62 rhizobial genomes using bioinformatics methods such as data mining in databases, sequence alignment, phenogram construction and protein modeling. Also, we analyzed soluble extracts from Bradyrhizobium japonicum USDA38 and USDA58 by (reduced + carbon monoxide (CO minus reduced differential spectroscopy. Database searching showed that only fhb, sdgb, gcs and thb genes exist in the rhizobia analyzed in this work. Promoter analysis revealed that apparently several rhizobial glb genes are not regulated by a -10 promoter but might be regulated by -35 and Fnr (fumarate-nitrate reduction regulator-like promoters. Mapping analysis revealed that rhizobial fhbs and thbs are flanked by a variety of genes whereas several rhizobial sdgbs and gcss are flanked by genes coding for proteins involved in the metabolism of nitrates and nitrites and chemotaxis, respectively. Phenetic analysis showed that rhizobial Glbs segregate into the M, S and T Glb families, while structural analysis showed that predicted rhizobial SDgbs and fHbs and GCSs globin domain and tHbs fold into the 3/3- and 2/2-folding, respectively. Spectra from B. japonicum USDA38 and USDA58 soluble extracts exhibited peaks and troughs characteristic of bacterial and vertebrate Glbs thus indicating that putative Glbs are synthesized in B. japonicum USDA38 and USDA58.

  1. Resistance to niclosamide in Oncomelania hupensis, the intermediate host of Schistosoma japonicum: should we be worried?

    Science.gov (United States)

    Dai, Jian-Rong; Li, You-Zi; Wang, Wei; Xing, Yun-Tian; Qu, Guo-Li; Liang, You-Sheng

    2015-02-01

    As the currently only available molluscicide, niclosamide has been widely used for snail control for over 2 decades in China. There is therefore a concern about the emergence of niclosamide-resistant snail populations following repeated, extensive use of the chemical. The purpose of this study was to investigate the likelihood of niclosamide resistance in Oncomelania hupensis in China. Active adult O. hupensis snails derived from 20 counties of 10 schistosomiasis-endemic provinces of China, of 10 snails in each drug concentration, were immersed in solutions of 1, 0.5, 0.25, 0.125, 0.063, 0.032, 0.016 and 0.008 mg L-1 of a 50% wettable powder of niclosamide ethanolamine salt (WPN) for 24 and 48 h at 25 °C, and the median lethal concentration (LC50) was estimated. Then, the 24- and 48-h WPN LC50 values were compared with those determined in the same sampling sites in 2002. The results indicated that the 24- and 48-h WPN LC50 values for O. hupensis were not significantly different from those determined in 2002 (P = 0.202 and 0.796, respectively). It is concluded that the current sensitivity of O. hupensis to niclosamide has not changed after more than 2 decades of repeated, extensive application in the main endemic foci of China, and there is no evidence of resistance to niclosamide detected in O. hupensis. PMID:25003984

  2. Establishment of Experimental Model of Hepatic Schistosoma Japonicum Egg Granulomas in Mice

    Institute of Scientific and Technical Information of China (English)

    冯振卿; 李玉华; 仇镇宁; 薛婉芬; 管晓虹

    1998-01-01

    IntroductionSchistosomaegggranulomasaretheprinci-palpathologicbasisofschistosomiasis.Toes-tablishanappropriateexperimentalmod...

  3. Study of the basis for the competitiveness of Rhizobium japonicum in the nodulation of soybean

    Energy Technology Data Exchange (ETDEWEB)

    Bauer, W.

    1990-06-01

    The overall goal of our studies was to identify cellular and molecular characteristics of rhizobia that are important to root colonization and symbiotic infection in competition with indigenous soil microbes. Rhizobia were found to respond to specific host flavonoids as chemoattractants as well as nodulation gene inducers. Motility and chemotaxis contributed significantly to the efficiency of symbiotic interactions, but not to colonization of root tips, where bacterial dispersal depended primarily on passive movement by root cell elongation. Co-inoculation of Rhizobium nod gene mutants with each other and with the wild-type showed that the mutants could help the wild-type, but not each other, to initiate nodules. Mutants with defective host specificity genes were the only ones capable of helping the wild-type in an additive or synergistic manner. Alfalfa was shown to have rapid, systemic feedback regulation suppressing nodule formation in younger parts of the root system in response to initiation of the first nodules. Root mucilage and exudate components were found to affect attachment of rhizobia to root surfaces. Methods for optimal surface sterilization of legume seeds were examined. 5 refs.

  4. Effect of artmether, hemin and Fe3+ on recombinant lactate dehydrogenase from Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    Gang Lu; Xuchu Hu; Can Huang; Yajun Lu; Lixian Wu; Lihua Li; Jing Xu; Xinbing Yu

    2010-01-01

    Objective: To explore antischitosome effects of artemether, hemin and Fe3+ on SjLDH. Methods:Enzyme activity of rSjLDH was assayed in the standard reaction system by adding different concentration of reagents (0.00-0.10 mM artemether, 0.00-0.02 mM hemin, 0.00-0.50 mM Fe3+). Same solvents of the each reagent were used as control. Results: There was no enzyme activity inhibition observed at 0.10 mM artemther; obivious inhibition for lactate oxidation reaction and pyruvate reduction reaction were detected at 0.002 mM and 0.004 mM of hemin, respectively;comparing with that of the control (P<0.05). The relative enzymatic activity inhibitions for pyruvate reduction reaction and lactate oxidation reaction at 0.02 mM hemin were 93.48% and 100.00%, respectively, comparing with that of the control (P<0.01); both pyruvate reduction and lactate oxidation reaction were inhibited completely at 0.50 mM Fe3+, comparing with that of the control (P<0.01). Conclusions: The results implied that SjLDH was not the direct molecular target of artemether. Hemin and Fe3+ are inhibitors of SjLDH.

  5. Isolation and Characterization of 44.6 kDa Protein from Schistosoma japonicum Male Worm

    Institute of Scientific and Technical Information of China (English)

    陈欲晓; 易新元; 曾宪芳; 袁仕善; 张顺科; LarryMCREYNOLDS

    2004-01-01

    Soluble male worm antigen of Schistosoma japonicurn (Sj) was investigated for development of new vaccine candidate. SDS--PAGE and Western blotting were performed to compare the difference between soluble antigens from worms of different sex. Mice vaccination with the testing purified protein was followed by Sj cereariae challenge to detect the protective effect against Sj. Sixteen bands were seen for the soluble male worm antigen and 12 for the female worm. In addition, a distinct band of 44.6 kDa from the male worm antigen was observed, and its antigenicity was demonstrated by Western blotting. This 44.6 kDa protein could induce significant worm and egg reduction rate in mice (39.31%, 41.98%, P < 0.001). Inthis study a 44.6 kDa protein was isolated and partially characterized. Its antigenicity, immunogenicity and the partial immune protection suggest its potential vaccine candidte against Sj.

  6. Local Antiglycan Antibody Responses to Skin Stage and Migratory Schistosomula of Schistosoma japonicum

    NARCIS (Netherlands)

    Smit, C.H.; Kies, C.L.; McWilliam, H.E.; Meeusen, E.N.; Hokke, C.H.; Diepen, A. van

    2015-01-01

    Schistosomiasis is a tropical disease affecting over 230 million people worldwide. Although effective drug treatment is available, reinfections are common, and development of immunity is slow. Most antibodies raised during schistosome infection are directed against glycans, some of which are thought

  7. Reconstitution of coronary vasculature by an active fraction of Geum japonicum in ischemic hearts

    Science.gov (United States)

    Chen, Hao; Cheng, Lei; Lin, Xiaoli; Zhou, Xiaping; Cai, Zhiming; Li, Ming

    2014-02-01

    Chronic coronary heart disease (cCHD) is characterized by atherosclerosis, which progressively narrows the coronary artery lumen and impairs myocardial blood flow. Restoration of occluded coronary vessels with newly formed collaterals remains an ideal therapeutic approach due to the need for redirecting blood flow into the ischemic heart. In this study, we investigated the effect of an active fraction isolated from Geum joponicum (AFGJ) on angiogenesis in cCHD hearts. Our results demonstrated that AFGJ not only enhanced capillary tube formation of endothelial cells, but also promoted the growth of new coronary collaterals (at the diameter 0.021-0.21 mm) in the ischemic region of hearts in rat cCHD model. Our study also indicated that the growth of new collaterals in ischemic hearts resulted in improved functional recovery of the cCHD hearts as demonstrated by ECG and echocardiography analyses. These data suggest that AFGJ may provide a novel therapeutic method for effective treatment of cCHD.

  8. Schistosoma japonicum:establishment of a mouse model that demonstrates concomitant immunity

    Institute of Scientific and Technical Information of China (English)

    HE Li 何立; YI Xin-yuan 易新元; JIANG Ming-sen 蒋明森; ZENG Xian-fang 曾宪芳; OUYANG Li 欧阳理; Larry McReynolds

    2004-01-01

    @@ Concomitant immunity to schistosome reinfection in vivo is generally thought to be the immune response induced by and relying on living adult worms.1,2 However, a few authors3,4 have suggested that the mechanism of resistance may be related to the hepatic pathology caused by the primary infection. The vascular pathology might enable the challenge schistosomula to escape from the hepatic portal system, thus preventing normal parasite sequestration and maturation in the liver.

  9. Crescimento de Bradyrhizobium elkanii estirpe Br 29 em meios de cultivo com diferentes valores de pH inicial Growth of Bradyrhizobium elkanii strain Br 29 in culture media with different pH values

    OpenAIRE

    Alexandre Barberi; Moreira, Fátima M. S.; Ligiane Aparecida Florentino; Maria Isabel D. Rodrigues

    2004-01-01

    A soja, atualmente, é a leguminosa de maior expressão econômica no Brasil. Em razão da demanda crescente no mercado internacional, a cultura encontra-se em larga expansão em todo território brasileiro. Em alguns ensaios realizados em solos ácidos brasileiros, sob sistema de plantio direto, essa cultura tem mostrado baixa resposta à aplicação de calcário. Porém, a acidez é um dos fatores que limitam a eficiência da simbiose rizóbio-leguminosas. Para contornar esses problemas, poderiam ser util...

  10. 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

  11. Physiological and Biochemical Responses to Drought Stress and Subsequent Rehydration in the Symbiotic Association Peanut-Bradyrhizobium sp.

    OpenAIRE

    Ana Furlan; Analía Llanes; Virginia Luna; Stella Castro

    2012-01-01

    Drought stress is one of the most important environmental factors that regulate plant growth and development and limit its production. Peanut (Arachis hypogaea L.) is an agriculturally valuable plant with widespread distribution in the world serving as a subsistence food crop as well as a source of various food products. The aims of this work were to evaluate growth and nodulation as well as some physiological and biochemical stress indicators in response to drought stress and subsequent rehy...

  12. 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.

  13. 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.

  14. Avaliação da atividade de microrganismos do solo em diferentes sistemas de manejo de soja Evaluation of nitrogen fixation and soil microorganisms in soybean under conventional and minimal cultivation regimes

    Directory of Open Access Journals (Sweden)

    O.M. de Castro

    1993-09-01

    Full Text Available Para avaliar o efeito do manejo do solo e de culturas sobre a atividade de Bradyrhizobium japonicum e de outros microrganismos importantes na ciclagem de nutrientes do solo, montou-se um experimento em casa de vegetação utilizando solo de um ensaio de campo com sistemas de manejo de resíduos e preparo bem distintos, como plantio direto e preparo convencional com arado de discos e diferentes rotações de cultura. O solo é um latossolo roxo distrófico, A moderado, textura argilosa. As culturas utilizadas no ensaio de campo foram soja contínua e em rotação com milho, sempre após aveia preta e Crolalaria juncea no outono-inverno, perfazendo portanto oito sistemas de manejo. Como testemunha, utilizou-se o solo de uma mata localizada próxima do experimento de campo. Prepararam-se seis vasos com solo de cada tratamento, sendo que em três foi semeada soja com sementes inoculadas com B. japonicum e nos outros três soja sem inoculação. No início do florescimento fez-se a colheita das plantas para análise de N na parte aérea, determinação dos nódulos e micorrização. Em amostras de solo de todos os tratamentos, guardadas em geladeira, fez-se a quantificação de bactérias e fungos totais, bem como de microrganismos celulolíticos, amonifícadores, nitritadores e nitratadores. Os resultados obtidos revelaram que o solo do plantio direto e o sistema de rotação levaram a significativos aumentos da nodulação. O solo do plantio direto apresentou também maiores populações de celulolíticos, não havendo diferença para outros tipos de microrganismos. Não houve diferença entre tratamentos para a micorrização, que apresentou valores muito baixos, provavelmente devido ao alto nível de fósforo no solo.A greenhouse experiment was set up to evaluate the effect of different cultural practices on the activity of Bradyrhizobium japonicum and of other soil microorganisms involved in nutrient cycling. The soil used was from a field

  15. 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

  16. 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.

  17. Use of arbitrary DNA primers, polyacrylamide gel electrophoresis and silver staining for identity testing, gene discovery and analysis of gene expression

    International Nuclear Information System (INIS)

    To understand chemically-induced genomic differences in soybean mutants differing in their ability to enter the nitrogen-fixing symbiosis involving Bradyrhizobium japonicum, molecular techniques were developed to aid the map-based, or positional, cloning. DNA marker technology involving single arbitrary primers was used to enrich regional RFLP linkage data. Molecular techniques, including two-dimensional pulse field gel electrophoresis, were developed to ascertain the first physical mapping in soybean, leading to the conclusion that in the region of marker pA-36 on linkage group H, 1 cM equals about 500 cM. High molecular weight DNA was isolated and cloned into yeast or bacterial artificial chromosomes (YACs/ BACs). YACs were used to analyze soybean genome structure, revealing that over half of the genome contains repetitive DNA. Genetic and molecular tools are now available to facilitate the isolation of plant genes directly involved in symbiosis. The further characterization of these genes, along with the determination of the mechanisms that lead to the mutation, will be of value to other plants and induced mutation research. (author)

  18. 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.

  19. 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

  20. 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.

  1. Structure and biological roles of Sinorhizobium fredii HH103 exopolysaccharide.

    Science.gov (United States)

    Rodríguez-Navarro, Dulce N; Rodríguez-Carvajal, Miguel A; 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

  2. Improvement of Rhizobial Inoculants: A Key Process in Sustainable Soybean Production

    Directory of Open Access Journals (Sweden)

    Mihaela Blažinkov

    2016-01-01

    Full Text Available Symbiotic nitrogen fixation has important role in sustainable soybean production because of utilization of atmospheric nitrogen for soybean nutrition. Pre-sowing soybean seed inoculation with selected rhizobial strains is used to improve the amount of symbiotically fixed nitrogen. Besides strain selection, suitable inoculant formulation is important for the success of inoculant application. The aim of this research is the evaluation of symbiotic efficiency and compatibility of Bradyrhizobium japonicum strains with soybean cultivar as well as possibility of using different inoculant formulation in soybean production. During two years of field trials in eastern Slavonia, nodule dry weight, nitrogen content in plant, seed yield, 1000 seeds weight, protein and oil content in seed were determined. Results of this study indicate that inoculant formulation as well as the use of selected strains affects nodulation, symbiotic and agronomic properties of soybean. Despite the differences in results in both experimental years, it can be concluded that the strains used as well as inoculant formulations are suitable for soybean inoculation in agroecological conditions of eastern Slavonia.

  3. 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.

  4. 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)

    LAZZARETTI E

    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.

  5. [Cloning and functional analysis of glnB from Azospirillum brasilense Yu62].

    Science.gov (United States)

    Li, Z H; Chen, S F; Li, J L

    2001-01-01

    The glnB gene of A. brasilense Yu62 was determined in a 3.7 kb EcoRI + PstI fragment. The glnA is located downstream of glnB and an ORF for hypothetical protein is on upstream of glnB. The deduced amino acid sequence of PII encoded by glnB is 71%, 77%, 79% and 69% identical to that of K. pneumoniae, Bradyrhizobium japonicum, Rhizobium leguninosarum and E. coli, respectively. A Km-casette was inserted into BglII site of glnB coding region and GlnB- mutant was obtained by homologous recombination. The GlnB- mutant has lost the nitrogenase activity, i.e.: Nif-. For the functional confirmation of glnB gene, a complementary test was carried out and it was shown that C-glnB(glnB::Km/glnB) can restore the nitrogenase activity. When the recombinant plasmid pVK-II which containined the coding region of glnB was introduced into A. brasilense Yu62 and A. brasilense Yu62 DraT-, respectively, the Yu62-II (containing pVK-II) and draT-II(containing pVK-II) showed higher nitrogenase activity than wild type. These results confirmed that glnB plays an important role in the regulation of nitrogen in A. brasilense.

  6. Complementation analyses of Sinorhizobium meliloti nifA mutant with different originated nifA genes

    Institute of Scientific and Technical Information of China (English)

    YAO Zhenhua; R(U)VERG Silvia; WANG Yiping; ZOU Huasong; TIAN Zhexian; DAI Xiaomi; BECKER Anke; LI Jian; YAN Haiqin; XIAO Yan; ZHU Jiabi; YU Guanqiao

    2006-01-01

    A previous work inferred that the nifA gene of Enterobacter cloacae did not restore the symbiotic phenotype of Sinorhizobium meliloti nifA mutant. In the present study, two nifA genes of Bradyrhizobium japonicum and Mesorhizobium huakuii also did not restore the symbiotic phenotype of S.meliloti nifA mutant. In whole genomic microarray experiments, 238 genes were found to be differentially expressed after S. meliloti nifA had been constitutively expressed in its nifA mutant. In contrast,only 20, 7 and 9 genes changed their transcriptional levels when expressing B. japonium, M. huakuii and Enterobacter cloacae nifA genes in Sm nifA mutant,separately. These genes were classified into several functional groups including house keeping, energy and central intermediary metabolism, transport systems and symbiosis. Interestingly, the genes that of nifH operons showed high expression levels in the presence of either B. japonium or M. huakuii NifA,which was confirmed by subsequent lacZ fusion experiments.

  7. Evolution of retinoic acid receptors in chordates: insights from three lamprey species, Lampetra fluviatilis, Petromyzon marinus, and Lethenteron japonicum

    OpenAIRE

    Campo-Paysaa, Florent; Jandzik, David; Takio-Ogawa, Yoko; Cattell, Maria V; Neef, Haley C; Langeland, James A.; Kuratani, Shigeru; Medeiros, Daniel M.; Mazan, Sylvie; Kuraku, Shigehiro; Laudet, Vincent; Schubert, Michael

    2015-01-01

    Background Retinoic acid (RA) signaling controls many developmental processes in chordates, from early axis specification to late organogenesis. The functions of RA are chiefly mediated by a subfamily of nuclear hormone receptors, the retinoic acid receptors (RARs), that act as ligand-activated transcription factors. While RARs have been extensively studied in jawed vertebrates (that is, gnathostomes) and invertebrate chordates, very little is known about the repertoire and developmental role...

  8. Molecular characterization and ligand binding specificity of the PDZ domain-containing protein GIPC3 from Schistosoma japonicum

    OpenAIRE

    Mu Yi; Huang Haiming; Liu Shuai; Cai Pengfei; Gao Youhe

    2012-01-01

    Abstract Background Schistosomiasis is a serious global health problem that afflicts more than 230 million people in 77 countries. Long-term mass treatments with the only available drug, praziquantel, have caused growing concerns about drug resistance. PSD-95/Dlg/ZO-1 (PDZ) domain-containing proteins are recognized as potential targets for the next generation of drug development. However, the PDZ domain-containing protein family in parasites has largely been unexplored. Methods We present the...

  9. 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.

  10. Gene : CBRC-ETEL-01-1376 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available 61 protein [Schistosoma japonicum] 2e-10 33% MPSRAKAQAYALAFPLPPCTMVFPPLPCTVVFLPLPCTMVFPPLPCTQRSLLSHAQWCSLFSHAQRCSLPLPCTMVFLPLPCT...MVVLSSSPXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXSPMHNSVALLSHVQWCSLLSPCTMIFLPLPCTMVFPPLPCTSLVCHAQWCSFLCLAQWCFLPLPCT...MVFPPLLFTMVFPPPLPCTTVFPPLPCTMVFPPLPCTMVFPPLPCTMVFPPLPCTMVFPRLPCTMVFPLLPCTMVFPPPLSCTMVFLPLPCTTVFPPLPYTIWYLGTQIILR ...

  11. Gene : CBRC-CJAC-01-0715 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available protein [Schistosoma japonicum] 3e-09 24% MRCYKYSTNTIPHYTDTLLLILIWCYKYSTNTIPHYTDTLLILILIWCYKYSTNTIPHYTETLLLILMRCYKYSTNT...IPYYTDTLLLILIWCYKYSTNTIPHYTDTLLILILIWCYKYSTNTIPHYTDTLLLILIRCYKYSTNTIPCYTETLLLLLYGATNTLLILYHTILIALLLILIWCYKYSTILXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXILY ...

  12. 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

    OpenAIRE

    Mana Kanjanamaneesathian; Paranee Sawangsri; Ashara Pengnoo; Jira Suwanprasert

    2007-01-01

    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 co...

  13. 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

  14. 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

  15. 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

  16. Development of a Microemulsion Formulation for Antimicrobial SecA Inhibitors

    Science.gov (United States)

    Wang, Nian

    2016-01-01

    In our previous study, we have identified five antimicrobial small molecules via structure based design, which inhibit SecA of Candidatus Liberibacter asiaticus (Las). SecA is a critical protein translocase ATPase subunit and is involved in pre-protein translocation across and integration into the cellular membrane in bacteria. In this study, eleven compounds were identified using similarity search method based on the five lead SecA inhibitors identified previously. The identified SecA inhibitors have poor aqueous solubility. Thus a microemulsion master mix (MMX) was developed to address the solubility issue and for application of the antimicrobials. MMX consists of N-methyl-2-pyrrolidone and dimethyl sulfoxide as solvent and co-solvent, as well as polyoxyethylated castor oil, polyalkylene glycol, and polyoxyethylene tridecyl ether phosphate as surfactants. MMX has significantly improved the solubility of SecA inhibitors and has no or little phytotoxic effects at concentrations less than 5.0% (v/v). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the SecA inhibitors and streptomycin against eight bacteria including Agrobacterium tumefaciens, Liberibacter crescens, Rhizobium etli, Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti phylogenetically related to Las were determined using the broth microdilution method. MIC and MBC results showed that the 16 SecA inhibitors have antibacterial activities comparable to that of streptomycin. Overall, we have identified 11 potent SecA inhibitors using similarity search method. We have developed a microemulsion formulation for SecA inhibitors which improved the antimicrobial activities of SecA inhibitors. PMID:26963811

  17. POST-EFFECT OF MYCORRHIZATION ON SOME INDICATORS OF PHYSICAL PROPERTIES FROM A RED FERRALITIC SOIL

    Directory of Open Access Journals (Sweden)

    M. C. Riera

    2003-01-01

    Full Text Available Se desarrolló un estudio en el área de experimentación del Instituto Nacional de Ciencias Agrícolas (INCA, provincia La Habana, sobre un suelo Ferralítico Rojo compactado, utilizando un diseño de parcelas divididas con cuatro repeticiones, para evaluar cuatro frecuencias de inoculación de HMA en dos secuencias de cultivos: soya (Glycine max. L-maíz (Zea mays-boniato (Ipomea batata L. y soyagirasol (Helianthus annus-sorgo (Sorghum vulgaris. La siembra se inició en la primavera de 1998 con 32 parcelas de 100 m2 (10 x 10 m, utilizando un área de cálculo de 56 m2. La aplicación de los biofertilizantes se realizó mediante el recubrimiento de las semillas antes de la siembra, siendo inoculada la soya con Bradyrhizobium japonicum (cepa ICA 8001 y Glomus clarum, mientras los restantes cultivos de cada secuencia se inocularon con Glomus clarum y Burkholderia cepacia. Para cada cultivo se realizaron evaluaciones del rendimiento, porcentaje de colonización micorrízica y la masa del endófito, mientras en el suelo se evaluaron la densidad real (Dr, densidad aparente (Da, textura, los micro y macroagregados, el coeficiente de estabilidad y el índice de dispersión. Los resultados mostraron una influencia marcada de las frecuencias de inoculación y de las secuencias de cultivos empleados sobre los macroagregados y otros indicadores del estado de las propiedades físicas del suelo.

  18. Absence of ancient DNA in sub-fossil insect inclusions preserved in 'Anthropocene' Colombian copal.

    Directory of Open Access Journals (Sweden)

    David Penney

    Full Text Available 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.

  19. 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

  20. 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.

  1. Subpicosecond oxygen trapping in the heme pocket of the oxygen sensor FixL observed by time-resolved resonance Raman spectroscopy.

    Science.gov (United States)

    Kruglik, Sergei G; Jasaitis, Audrius; Hola, Klara; Yamashita, Taku; Liebl, Ursula; Martin, Jean-Louis; Vos, Marten H

    2007-05-01

    Dissociation of oxygen from the heme domain of the bacterial oxygen sensor protein FixL constitutes the first step in hypoxia-induced signaling. In the present study, the photodissociation of the heme-O2 bond was used to synchronize this event, and time-resolved resonance Raman (TR(3)) spectroscopy with subpicosecond time resolution was implemented to characterize the heme configuration of the primary photoproduct. TR(3) measurements on heme-oxycomplexes are highly challenging and have not yet been reported. Whereas in all other known six-coordinated heme protein complexes with diatomic ligands, including the oxymyoglobin reported here, heme iron out-of-plane motion (doming) occurs faster than 1 ps after iron-ligand bond breaking; surprisingly, no sizeable doming is observed in the oxycomplex of the Bradyrhizobium japonicum FixL sensor domain (FixLH). This assessment is deduced from the absence of the iron-histidine band around 217 cm(-1) as early as 0.5 ps. We suggest that efficient ultrafast oxygen rebinding to the heme occurs on the femtosecond time scale, thus hindering heme doming. Comparing WT oxy-FixLH, mutant proteins FixLH-R220H and FixLH-R220Q, the respective carbonmonoxy-complexes, and oxymyoglobin, we show that a hydrogen bond of the terminal oxygen atom with the residue in position 220 is responsible for the observed behavior; in WT FixL this residue is arginine, crucially implicated in signal transmission. We propose that the rigid O2 configuration imposed by this residue, in combination with the hydrophobic and constrained properties of the distal cavity, keep dissociated oxygen in place. These results uncover the origin of the "oxygen cage" properties of this oxygen sensor protein.

  2. pHairyRed: A Novel Binary Vector Containing the DsRed2 Reporter Gene for Visual Selection of Transgenic Hairy Roots

    Institute of Scientific and Technical Information of China (English)

    Meng-Han Lin; Peter M. Gresshoff; Arief Indrasumunar; Brett J. Ferguson

    2011-01-01

    T We developed a new plant transformation vector, pHairyRed, for enabling high throughput, non-destructive selection of Agrobacterium rhizogenes-mediated 'hairy-root' transformation. pHairyRed allows easy in planta visualization of transgenic tissue with minimal disturbance to the plant. The DsRed2 reporter gene, encoding a red fluorescent protein, was cloned to yield pHairyRed (harbouring a multiple cloning site), which was used with the highly efficient K599 A. rhizogenes strain to infect soybean (G/ycine max L. Merrill) plants. DsRed2 fluorescence was easily detected in planta for the duration of a 5-week study with negligible levels of background autofluorescence. This enabled visual selection of transformed roots and subsequent excission of non-transformed roots. pHairyRed-transformed roots nodulated normally when inoculated with Bradyrhizobium japonicum. Within the nodule, DsRed2 fluorescence was plant-specific, being absent in the bacteroid-dominated nodule infected zone. To test the reliability of pHairyRed as a high-fidelity binary vector reporter system, the gene encoding the soybean Nod factor receptor, GmNFR1α, was cloned into the vector for use in a complementation study with a non-nodulating nfr1α mutant of soybean. Complementation was achieved and, without exception, DsRed2 fluorescence was detected in all hairy roots that successfully formed nodules (100%, n = 34).We anticipate broad application of this reporter system for the further analysis of root-related events in soybean and related legumes.

  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. Functional analysis of the cysteine motifs in the ferredoxin-like protein FdxN of Rhizobium meliloti involved in symbiotic nitrogen fixation.

    Science.gov (United States)

    Masepohl, B; Kutsche, M; Riedel, K U; Schmehl, M; Klipp, W; Pühler, A

    1992-05-01

    The Rhizobium meliloti fdxN gene, which is part of the nifA-nifB-fdxN operon, is absolutely required for symbiotic nitrogen fixation. The deduced sequence of the FdxN protein is characterized by two cysteine motifs typical of bacterial-type ferredoxins. The Fix-phenotype of an R. meliloti fdxN::[Tc] mutant could be rescued by the R. leguminosarum fdxN gene, whereas no complementation was observed with nif-associated genes encoding ferredoxins from Bradyrhizobium japonicum, Azotobacter vinelandii, A. chroococcum and Rhodobacter capsulatus. In addition to these heterologous genes, several R. meliloti fdxN mutant genes constructed by site-directed mutagenesis were analyzed. Not only a cysteine residue within the second cysteine motif (position 42), which is known to coordinate the Fe-S cluster in homologous proteins, but also a cysteine located down-stream of this motif (position 61), was found to be essential for the activity of the R. meliloti FdxN protein. Changing the amino acid residue proline in position 56 into methionine resulted in a FdxN mutant protein with decreased activity, whereas changes in positions 35 (Asp35Glu) and 45 (Gly45Glu) had no significant effect on the function of the FdxN mutant proteins. In contrast to bacterial-type ferredoxins, which contain two identical cysteine motifs of the form C-X2-C-X2-C-X3-C, nif-associated ferredoxins, including R. meliloti FdxN, are characterized by two different cysteine motifs. Six "additional" amino acids separate the second (Cys42) and the third cysteine (Cys51) in the C-terminal motif (C-X2-C-X8-C-X3-C).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1603075

  5. Subpicosecond oxygen trapping in the heme pocket of the oxygen sensor FixL observed by time-resolved resonance Raman spectroscopy.

    Science.gov (United States)

    Kruglik, Sergei G; Jasaitis, Audrius; Hola, Klara; Yamashita, Taku; Liebl, Ursula; Martin, Jean-Louis; Vos, Marten H

    2007-05-01

    Dissociation of oxygen from the heme domain of the bacterial oxygen sensor protein FixL constitutes the first step in hypoxia-induced signaling. In the present study, the photodissociation of the heme-O2 bond was used to synchronize this event, and time-resolved resonance Raman (TR(3)) spectroscopy with subpicosecond time resolution was implemented to characterize the heme configuration of the primary photoproduct. TR(3) measurements on heme-oxycomplexes are highly challenging and have not yet been reported. Whereas in all other known six-coordinated heme protein complexes with diatomic ligands, including the oxymyoglobin reported here, heme iron out-of-plane motion (doming) occurs faster than 1 ps after iron-ligand bond breaking; surprisingly, no sizeable doming is observed in the oxycomplex of the Bradyrhizobium japonicum FixL sensor domain (FixLH). This assessment is deduced from the absence of the iron-histidine band around 217 cm(-1) as early as 0.5 ps. We suggest that efficient ultrafast oxygen rebinding to the heme occurs on the femtosecond time scale, thus hindering heme doming. Comparing WT oxy-FixLH, mutant proteins FixLH-R220H and FixLH-R220Q, the respective carbonmonoxy-complexes, and oxymyoglobin, we show that a hydrogen bond of the terminal oxygen atom with the residue in position 220 is responsible for the observed behavior; in WT FixL this residue is arginine, crucially implicated in signal transmission. We propose that the rigid O2 configuration imposed by this residue, in combination with the hydrophobic and constrained properties of the distal cavity, keep dissociated oxygen in place. These results uncover the origin of the "oxygen cage" properties of this oxygen sensor protein. PMID:17446273

  6. Prokaryotic squalene-hopene cyclases can be converted to citronellal cyclases by single amino acid exchange.

    Science.gov (United States)

    Siedenburg, Gabriele; Breuer, Michael; Jendrossek, Dieter

    2013-02-01

    Squalene-hopene cyclases (SHCs) are prokaryotic enzymes that catalyse the cyclisation of the linear precursor squalene to pentacyclic hopene. Recently, we discovered that a SHC cloned from Zymomonas mobilis (ZMO-1548 gene product) has the unique property to cyclise the monoterpenoid citronellal to isopulegol. In this study, we performed saturation mutagenesis of three amino acids of the catalytic centre of ZMO-1548 (F428, F486 and W555), which had been previously identified to interact with enzyme-bound substrate. Replacement of F428 by tyrosine increased hopene formation from squalene, but isopulegol-forming activity was strongly reduced or abolished in all muteins of position 428. W555 was essential for hopene formation; however, three muteins (W555Y, W428F or W555T) revealed enhanced cyclisation efficiency with citronellal. The residue at position 486 turned out to be the most important for isopulegol-forming activity. While the presence of phenylalanine or tyrosine favoured cyclisation activity with squalene, several small and/or hydrophobic residues such as cysteine, alanine or isoleucine and others reduced activity with squalene but greatly enhanced isopulegol formation from citronellal. Replacement of the conserved aromatic residue corresponding to F486 to cysteine in other SHCs cloned from Z. mobilis (ZMO-0872), Alicyclobacillus acidocaldarius (SHC(Aac)), Acetobacter pasteurianus (SHC(Apa)), Streptomyces coelicolor (SHC(Sco)) and Bradyrhizobium japonicum (SHC(Bja)) resulted in more or less strong isopulegol-forming activities from citronellal. In conclusion, many SHCs can be converted to citronellal cyclases by mutagenesis of the active centre thus broadening the applicability of this interesting class of biocatalyst. PMID:22526778

  7. Development of a Microemulsion Formulation for Antimicrobial SecA Inhibitors.

    Directory of Open Access Journals (Sweden)

    Jiahuai Hu

    Full Text Available In our previous study, we have identified five antimicrobial small molecules via structure based design, which inhibit SecA of Candidatus Liberibacter asiaticus (Las. SecA is a critical protein translocase ATPase subunit and is involved in pre-protein translocation across and integration into the cellular membrane in bacteria. In this study, eleven compounds were identified using similarity search method based on the five lead SecA inhibitors identified previously. The identified SecA inhibitors have poor aqueous solubility. Thus a microemulsion master mix (MMX was developed to address the solubility issue and for application of the antimicrobials. MMX consists of N-methyl-2-pyrrolidone and dimethyl sulfoxide as solvent and co-solvent, as well as polyoxyethylated castor oil, polyalkylene glycol, and polyoxyethylene tridecyl ether phosphate as surfactants. MMX has significantly improved the solubility of SecA inhibitors and has no or little phytotoxic effects at concentrations less than 5.0% (v/v. The minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC of the SecA inhibitors and streptomycin against eight bacteria including Agrobacterium tumefaciens, Liberibacter crescens, Rhizobium etli, Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti phylogenetically related to Las were determined using the broth microdilution method. MIC and MBC results showed that the 16 SecA inhibitors have antibacterial activities comparable to that of streptomycin. Overall, we have identified 11 potent SecA inhibitors using similarity search method. We have developed a microemulsion formulation for SecA inhibitors which improved the antimicrobial activities of SecA inhibitors.

  8. Development of a Microemulsion Formulation for Antimicrobial SecA Inhibitors.

    Science.gov (United States)

    Hu, Jiahuai; Akula, Nagaraju; Wang, Nian

    2016-01-01

    In our previous study, we have identified five antimicrobial small molecules via structure based design, which inhibit SecA of Candidatus Liberibacter asiaticus (Las). SecA is a critical protein translocase ATPase subunit and is involved in pre-protein translocation across and integration into the cellular membrane in bacteria. In this study, eleven compounds were identified using similarity search method based on the five lead SecA inhibitors identified previously. The identified SecA inhibitors have poor aqueous solubility. Thus a microemulsion master mix (MMX) was developed to address the solubility issue and for application of the antimicrobials. MMX consists of N-methyl-2-pyrrolidone and dimethyl sulfoxide as solvent and co-solvent, as well as polyoxyethylated castor oil, polyalkylene glycol, and polyoxyethylene tridecyl ether phosphate as surfactants. MMX has significantly improved the solubility of SecA inhibitors and has no or little phytotoxic effects at concentrations less than 5.0% (v/v). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the SecA inhibitors and streptomycin against eight bacteria including Agrobacterium tumefaciens, Liberibacter crescens, Rhizobium etli, Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti phylogenetically related to Las were determined using the broth microdilution method. MIC and MBC results showed that the 16 SecA inhibitors have antibacterial activities comparable to that of streptomycin. Overall, we have identified 11 potent SecA inhibitors using similarity search method. We have developed a microemulsion formulation for SecA inhibitors which improved the antimicrobial activities of SecA inhibitors. PMID:26963811

  9. Evaluation of the tepary bean (Phaseolus acutifolius) diversity panel for response to the NL 3 strain of Bean Common Mosaic Necrosis Virus (BCMNV) and for biological nitrogen fixation with Bradyrhizobium strains

    Science.gov (United States)

    Aphid-transmitted Bean Common Mosaic Necrosis Virus (BCMNV) and Bean Common Mosaic Virus (BCMV) are potyviruses that are seed transmitted in tepary bean. Developing resistance to these viruses will be critical for expanding production in areas where they are endemic. Biological nitrogen fixation (BN...

  10. Reintroducción del altramuz endémico valenciano Lupinus mariaejosephae mediante la inoculación de cepas seleccionadas de bacterias del género Bradyrhizobium

    OpenAIRE

    Albert NAVARRO; Oltra Benavent, Josep E.; Duran Wendt, David Ricardo; Rey Navarro, Luis; Sanchez Cañizares, Carmen; Ruiz Argüeso, Tomas-Andres; Imperial Ródenas, Juan; Emilio LAGUNA LUMBRERAS; Escribá, Mari Carmen; Albert, Francisco J.; Ferrando, Inmaculada; Ferrer, Pedro Pablo (S.I.); Fos, Simón

    2013-01-01

    Lupinus mariae-josephae es un altramuz endémico de la provincia de Valencia de reciente descubrimiento para la ciencia. Se conocen solo 4 poblaciones, algunas con miles de individuos pero todas ellas con grandes fluctuaciones interanuales, tanto demográficas como de éxito reproductivo. Es por ello que está incluida en el Catálogo Valenciano de Especies de Flora Amenazadas como "Especie Vulnerable". Con finalidad conservacionista se realizó una reintroducción de la especie dentro de su área de...

  11. Phylogenomics and signature proteins for the alpha Proteobacteria and its main groups

    Directory of Open Access Journals (Sweden)

    Mok Amy

    2007-11-01

    Full Text Available Abstract Background Alpha proteobacteria are one of the largest and most extensively studied groups within bacteria. However, for these bacteria as a whole and for all of its major subgroups (viz. Rhizobiales, Rhodobacterales, Rhodospirillales, Rickettsiales, Sphingomonadales and Caulobacterales, very few or no distinctive molecular or biochemical characteristics are known. Results We have carried out comprehensive phylogenomic analyses by means of Blastp and PSI-Blast searches on the open reading frames in the genomes of several α-proteobacteria (viz. Bradyrhizobium japonicum, Brucella suis, Caulobacter crescentus, Gluconobacter oxydans, Mesorhizobium loti, Nitrobacter winogradskyi, Novosphingobium aromaticivorans, Rhodobacter sphaeroides 2.4.1, Silicibacter sp. TM1040, Rhodospirillum rubrum and Wolbachia (Drosophila endosymbiont. These studies have identified several proteins that are distinctive characteristics of all α-proteobacteria, as well as numerous proteins that are unique repertoires of all of its main orders (viz. Rhizobiales, Rhodobacterales, Rhodospirillales, Rickettsiales, Sphingomonadales and Caulobacterales and many families (viz. Rickettsiaceae, Anaplasmataceae, Rhodospirillaceae, Acetobacteraceae, Bradyrhiozobiaceae, Brucellaceae and Bartonellaceae. Many other proteins that are present at different phylogenetic depths in α-proteobacteria provide important information regarding their evolution. The evolutionary relationships among α-proteobacteria as deduced from these studies are in excellent agreement with their branching pattern in the phylogenetic trees and character compatibility cliques based on concatenated sequences for many conserved proteins. These studies provide evidence that the major groups within α-proteobacteria have diverged in the following order: (Rickettsiales(Rhodospirillales (Sphingomonadales (Rhodobacterales (Caulobacterales-Parvularculales (Rhizobiales. We also describe two conserved inserts in DNA

  12. АГРОТЕХНОЛОГИЧЕСКИЕ ОСНОВЫ СОЗДАНИЯ УСОВЕРШЕНСТВОВАННЫХ ФОРМ МИКРОБНЫХ БИОПРЕПАРАТОВ ДЛЯ ЗЕМЛЕДЕЛИЯ

    OpenAIRE

    Кожемяков, А.; Лактионов, Ю.; Попова, Т.; Орлова, А.; Кокорина, А.; Вайшля, О.; Агафонов, Е.; Гужвин, С.; Чураков, А.; Яковлева, М.

    2015-01-01

    Выполнены комплексные исследования по созданию жидкой формы биопрепаратов для симбиотических и ассоциативных ризобактерий. Объектами изучения служили клубеньковые бактерии, обитающие в ризосфере козлятника (Rhizobium galegae), сои (Bradyrhizobium japonicum), и ассоциативные ризобактерии (Arthrobacter mysorens, Azospirillum brasilense, Ag-robacterium radiobacter), а также растения сои (Glycine max), кукурузы (Zea L.) и пшеницы (Triticum L.). В качестве носителей бактерий использовали гамма-сте...

  13. The effect of mulching and soil compaction on fungi composition and microbial communities in the rhizosphere of soybean

    Science.gov (United States)

    Frac, M.; Siczek, A.; Lipiec, J.

    2009-04-01

    The soil environment is the habitat of pathogenic and saprotrophic microorganisms. The composition of the microbial community are related to biotic and abiotic factors, such as root exudates, crop residues, climate factors, mulching, mineral fertilization, pesticides introduction and soil compaction. The aim of the study was to determine the effect of the mulching and soil compaction on the microorganism communities in the rhizosphere soil of soybean. The studies were carried out on silty loam soil (Orthic Luvisol) developed from loess (Lublin, Poland). The experiment area was 192m2 divided into 3 sections consisted of 6 micro-plots (7m2). Three levels of soil compaction low, medium and heavy obtained through tractor passes were compared. The soil was compacted and loosened within seedbed layer 2 weeks before sowing. Soybean "Aldana" seeds were inoculated with Bradyrhizobium japonicum and were sown with interrow spacing of 0.3m. Wheat straw (as mulch) was uniformly spread on the half of each micro-plot at an amount of 0.5kg m-1 after sowing. Rhizosphere was collected three times during growing season of soybean. Microbiological analyses were conducted in 3 replications and included the determination of: the total number of bacteria and fungi, the number of bacteria Pseudomonas sp. and Bacillus sp., the genus identification of fungi isolated from rhizosphere of soybean. Results indicated a positive effect of mulching on the increase number of all groups of examined rhizosphere microorganisms (fungi, bacteria, Pseudomonas sp., Bacillus sp.). The highest number of the microorganisms was found in the low and medium compacted soil and markedly decreased in the most compacted soil. Relatively high number of antagonistic fungi (Penicillium sp., Trichoderma sp.) was recorded in the rhizosphere of low and medium compacted soil, particularly in mulched plots. The presence of these fungi can testify to considerable biological activity, which contributes to the improvement of

  14. The tep1 gene of Sinorhizobium meliloti coding for a putative transmembrane efflux protein and N-acetyl glucosamine affect nod gene expression and nodulation of alfalfa plants

    Directory of Open Access Journals (Sweden)

    Soto María

    2009-01-01

    Full Text Available Abstract Background Soil bacteria collectively known as Rhizobium, characterized by their ability to establish beneficial symbiosis with legumes, share several common characteristics with pathogenic bacteria when infecting the host plant. Recently, it was demonstrated that a fadD mutant of Sinorhizobium meliloti is altered in the control of swarming, a type of co-ordinated movement previously associated with pathogenicity, and is also impaired in nodulation efficiency on alfalfa roots. In the phytopathogen Xanthomonas campestris, a fadD homolog (rpfB forms part of a cluster of genes involved in the regulation of pathogenicity factors. In this work, we have investigated the role in swarming and symbiosis of SMc02161, a S. meliloti fadD-linked gene. Results The SMc02161 locus in S. meliloti shows similarities with members of the Major Facilitator Superfamily (MFS of transporters. A S. meliloti null-mutant shows increased sensitivity to chloramphenicol. This indication led us to rename the locus tep1 for transmembrane efflux protein. The lack of tep1 does not affect the appearance of swarming motility. Interestingly, nodule formation efficiency on alfalfa plants is improved in the tep1 mutant during the first days of the interaction though nod gene expression is lower than in the wild type strain. Curiously, a nodC mutation or the addition of N-acetyl glucosamine to the wild type strain lead to similar reductions in nod gene expression as in the tep1 mutant. Moreover, aminosugar precursors of Nod factors inhibit nodulation. Conclusion tep1 putatively encodes a transmembrane protein which can confer chloramphenicol resistance in S. meliloti by expelling the antibiotic outside the bacteria. The improved nodulation of alfalfa but reduced nod gene expression observed in the tep1 mutant suggests that Tep1 transports compounds which influence nodulation. In contrast to Bradyrhizobium japonicum, we show that in S. meliloti there is no feedback regulation

  15. Genome organization and characteristics of soybean microRNAs

    Directory of Open Access Journals (Sweden)

    Turner Marie

    2012-05-01

    Full Text Available Abstract Background microRNAs (miRNAs are key regulators of gene expression and play important roles in many aspects of plant biology. The role(s of miRNAs in nitrogen-fixing root nodules of leguminous plants such as soybean is not well understood. We examined a library of small RNAs from Bradyrhizobium japonicum-inoculated soybean roots and identified novel miRNAs. In order to enhance our understanding of miRNA evolution, diversification and function, we classified all known soybean miRNAs based on their phylogenetic conservation (conserved, legume- and soybean-specific miRNAs and examined their genome organization, family characteristics and target diversity. We predicted targets of these miRNAs and experimentally validated several of them. We also examined organ-specific expression of selected miRNAs and their targets. Results We identified 120 previously unknown miRNA genes from soybean including 5 novel miRNA families. In the soybean genome, genes encoding miRNAs are primarily intergenic and a small percentage were intragenic or less than 1000 bp from a protein-coding gene, suggesting potential co-regulation between the miRNA and its parent gene. Difference in number and orientation of tandemly duplicated miRNA genes between orthologous genomic loci indicated continuous evolution and diversification. Conserved miRNA families are often larger in size and produce less diverse mature miRNAs than legume- and soybean-specific families. In addition, the majority of conserved and legume-specific miRNA families produce 21 nt long mature miRNAs with distinct nucleotide distribution and regulate a more conserved set of target mRNAs compared to soybean-specific families. A set of nodule-specific target mRNAs and their cognate regulatory miRNAs had inverse expression between root and nodule tissues suggesting that spatial restriction of target gene transcripts by miRNAs might govern nodule-specific gene expression in soybean. Conclusions Genome

  16. Soybean after the consortium corn with tropical forages in the savannah

    Directory of Open Access Journals (Sweden)

    Cássia Maria de Paula Garcia

    2013-12-01

    Full Text Available The biggest limitation to sustainable tillage in the Savannah region is the low biomass accumulation during autumn up to spring, due to low water availability and high temperatures characterizing these regions as having a warm and dry winter. The consortium of grains crops with tropical forages mainly of the genus Panicum and Urochloa is an alternative, to this situation. This study aimed to evaluate the effect of the straw production of Panicum and Urochloa after consortium with corn in the crop-livestock integration system on the soybean crop irrigated in succession. The experiment was conducted at the Farm for Teaching, Research and Extension, Faculty of Engineering - UNESP, Ilha Solteira in an Oxisol in Savannah conditions, being in no-tillage for 8 years (previous corn crop. The experimental design was a randomized block with four replications and four treatments: Panicum maximum cv. Tanzania sown during the nitrogen fertilization (CTD of the corn; Panicum maximum cv. Mombasa sown during the nitrogen fertilization (CMD of the corn; Urochloa brizantha cv. Xaraés sown during the occasion of nitrogen fertilization (CBD of the corn; Urochloa ruziziensis cv. Comum sown during the nitrogen fertilization (CRD of the corn. After harvesting the corn and forages cuts, soybean was planted with seeds inoculated with Bradyrhizobium japonicum (Turfy with 600,000 viable cells/seed, at distances of 0.45 m with 24 seeds-1 in the no tillage system (NTS. The height of the first pod, plant population and yield components of soybean (number of pods per plant, number of seeds per pod and mass of 100 grains and grain yield of soybean were not significantly influenced by the straws of forages. These results were found, probably because the area already has a history of tillage for 8 years since this system was stabilized in the area. Therefore, in this soil that was managed correctly, the straw production of the forage of genus Panicum and Urochloa does not

  17. Isolation and application of effective nitrogen fixation rhizobial strains on low-phosphorus acid soils in South China

    Institute of Scientific and Technical Information of China (English)

    CHENG FengXian; CAO GuiQin; WANG XiuRong; ZHAO Jing; YAN XiaoLong; LIAO Hong

    2009-01-01

    Soybean (Glycine max L.) is a very important food and oil crop in China. Legume-rhizobium symbiotic nitrogen (N) fixation is an important biological character and also the base of improving soil fertility of soybean. However, soybean production and development is severely limited in tropical and subtropical areas in China due to a lack of effective rhizobial inoculants adapting to low-phosphorus (P) acid soils. In the present study, 12 soybean rhizobial strains were isolated and purified from the nodules of two soybean genotypes contrasting in P efficiency, which were grown on different Iow-P acid soils with different soybean cultivation histories. Results from 16S rDNA sequence analysis showed that these 12 rhizobial strains belonged to the genus of Bradyrhizobium, which had higher nitrogenase activities compared to the control strain, Bradyrhizboium japonicum USDA110. A field experiment was carried out by applying rhizobial inoculants, a mixture of three rhizobial strains that showed the highest ni-trogenase activity, on a typical Iow-P acid soil in South China. The results showed that, without inocu-lation, no nodules were formed in the three soybean genotypes tested; with inoculation, the nodulation rates in all were 100%. Inoculation with rhizobial inocuiants not only made many nodules formed, but also increased soybean shoot biomass and yield, and improved nitrogen (N) and P nutrient status. Among which, shoot dry weight, N and P content of a soybean genotype, Huachun 3, inoculated with rhizobium were increased 154.3%, 152.4% and 163.2% compared to that without inoculation, respec-tively. We concluded that: (i) The effective indigenous rhizobial strains isolated in this study from soybeans on Iow-P acid soils in South China have the characters of broad host range, high nodulation efficiency, efficient N fixation, great low pH and low P tolerance. (ii) Soil environment and host types are the key factors to screen the effective rhizobial strains. Considering

  18. ADDITION OF PEARL MILLET STRAW TO SOIL FOR REDUCTION OF SOUTHERN BLIGHT IN COMMOM BEAN ADIÇÃO DE PALHA DE MILHETO AO SOLO PARA REDUÇÃO DA PODRIDÃO DO COLO EM FEIJOEIRO

    Directory of Open Access Journals (Sweden)

    João Vicente Pereira Neto

    2010-08-01

    Full Text Available

    Many plants, such as pearl millet (Pennisetum glaucum, have been cultivated with the purpose of covering or being incorporated to soil. This practice might improve soil chemical, physical and biological characteristics. These changes in soil features can affect diseases, especially those caused by soil-borne pathogens, such as Sclerotium rolfsii. In this study, a group of experiments was conducted in a randomized block design to evaluated the effects of pearl millet straw addition (N = 0.8%; P = 0.14%; K = 1.76%; Ca = 0.52%; Mg = 0.22%; S = 0.12%; B = 6.8 mg kg-1; Cu = 12.7 mg kg-1; Fe = 412 mg kg-1; Mn = 39.5 mg kg-1; Zn = 30.3 mg kg-1; and Na = 360 mg kg-1 and seed inoculation with Bradyrhizobium japonicum on common bean (Phaseolus vulgaris southern blight, cultivated in a greenhouse, and also on the fluorescent Pseudomonas spp. native population. The addition of straw (10 t ha-1, 20 t ha-1, and 30 t ha-1 reduced the severity of the disease, while the seed inoculation with Rhizobium did not. There was a significant increase of fluorescent Pseudomonas populations in soils with straw. These results indicate that the addition of pearl millet straw to soil can reduce the disease and favour the fluorescent Pseudomonas population growth.

    KEY-WORDS: Sclerotium rolfsii; Pennisetum glaucum; Phaseolus vulgaris; fluorescent

  19. Research progress on Sj14-3-3 vaccine of Schistosoma japonicum%日本血吸虫Sj14-3-3疫苗研究进展

    Institute of Scientific and Technical Information of China (English)

    谭建蓉; 李文桂

    2014-01-01

    日本血吸虫病是由日本血吸虫引起的一类严重危害人类健康的人兽共患寄生虫病,研制疫苗防治该病是目前的研究热点.Sj14-3-3蛋白是一种有效的疫苗分子,该文就Sj14-3-3蛋白疫苗和核酸疫苗的研究进展进行综述.%Schistosomiasis japonica is a serious health-threatening parasitic zoonosis to human beings,which is caused by Schistosomajaponicum.Developing vaccines for schistosomiasis is a hot spot in the present studies.Sj14-3-3 protein is an effective vaccine.This article reviewed the progress on Sj14-3-3 protein vaccines and DNA vaccines.

  20. 慢生大豆根瘤菌与竞争结瘤相关的Irp基因的克隆%Cloning of Irp Gene Related to Nodule Competitiveness in Bradyrhizoblum Japonicum

    Institute of Scientific and Technical Information of China (English)

    武波; 唐成来; 柏学亮; 唐东阶; 吕安国; 唐纪良; 马庆生

    2002-01-01

    通过对重组质粒pGXN300中的2.3kb EcoRI片段测序分析发现,其中含有一完整的lrp基因和部分putA基因,与King等报道的B.japonicum的lrp基因DNA序列有88%同源性.应用Tn5gusA5定位诱变的方法获得了gusA基因表达的根瘤菌lrp基因突变体GX20108.植株试验表明,突变株结瘤时间明显推迟,竞争结瘤能力也显著下降.

  1. 日本血吸虫成虫肠道内容物中金属蛋白酶的鉴定%Identification of Metalloproteinase in Intestinal Contents Of Adult Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    袁仕善; 易新元; 曾宪芳; 张顺科; 黄跃龙; Larry MeReynolds

    2002-01-01

    目的鉴定日本血吸虫成虫肠道内容物中的金属蛋白酶.方法以明胶作底物,利用明胶-SDS-PAGE分离日本血吸虫成虫肠道内容物,将电泳后的凝胶于不同pH缓冲液和酶抑制剂中进行孵育,对其中的金属蛋白酶进行分析和鉴定.结果日本血吸虫成虫肠道内容物中存在降解明胶的金属蛋白酶,其最适pH值为7~9.金属蛋白酶抑制剂EDTA抑制其活性.电泳分离获得有活性的酶蛋白,该酶是血吸虫感染血清识别的弱抗原.结论日本血吸虫成虫肠道内容物存在金属蛋白酶.

  2. Construction and analysis of Sj - Rho GTPase DNA vaccine and subunit vaccine against Schistosoma japonicum%日本血吸虫Rho GTPase DNA疫苗及亚单位疫苗的构建和分析

    Institute of Scientific and Technical Information of China (English)

    张冉; 易新元; 曾宪芳; 黄跃龙; 蔡春; 张顺科; Larry McReynolds

    2003-01-01

    目的构建日本血吸虫大陆株Sj-Rho GTPase-like真核及原核表达重组质粒,并进行鉴定分析.方法用PCR法将Si-Rho GTPase-like基因从已剪切阳性克隆中扩增出来,亚克隆至pcDNA3.1和pGEX-5X-3中,分别经PCR、双酶切、测序、SDS-PAGE和Western blot等方法鉴定.结果PCR和测序均证明Sj-Rho GTPase-like基因疫苗构建成功,重组蛋白经SDS-PAGE电泳可观察到与预期分子量相应的条带,转印后可被水牛感染血清识别.结论成功构建了Si-Rho GTPase-like基因的两种重组载体,为保护性免疫研究打下基础.

  3. STUDY ON PROTECTION AGAINST SCHISTOSOMA JAPONICUM INDUCED BYPASSIVELY TRANSFERED SERA FROM MICROTUS FOTIS IN THE MOUSE%东方田鼠血清被动转移抗日本血吸虫的保护力研究

    Institute of Scientific and Technical Information of China (English)

    蒋守富; 魏梅雄; 林矫矫; 潘彩娥; 李浩; 曹琳; 何艳燕; 傅志强

    2004-01-01

    目的探讨被动转移东方田鼠血清抗日本血吸虫感染力及其作用机理. 方法将东方田鼠血清通过尾静脉注射途径被动转移至小鼠,观察攻击感染日本血吸虫尾蚴后的减虫率、减卵率,并采用ELISA分别检测抗日本血吸虫童虫、成虫和虫卵的8种抗体. 结果与生理盐水对照组比较, 东方田鼠血清受体小鼠获得的减虫率为36.2%,减卵率为54.0%;血清IgE、IgM、IgG及其亚类抗体均有升高,其中抗童虫抗原的IgG1抗体水平增幅最大.各试验组小鼠血清抗体水平差异均无显著性(P>0.05). 结论东方田鼠抗日本血吸虫感染的天然抵抗力可通过血清被动转移至小鼠,使之获得部分抗血吸虫感染的保护力,该保护力可能是通过同时诱导受体小鼠Th1和Th2型免疫应答发挥作用的.

  4. EFFECT OF pH ON THE MUTAGENIC POTENCY OF ICR-170 IN THE REPRODUCTIVE GLANDS OF FEMALE SCHISTOSOMA JAPONICUM%PH对ICR-170诱变日本血吸虫雌虫生殖腺的影响

    Institute of Scientific and Technical Information of China (English)

    蒋守富; 潘彩娥; 陆钦淹

    2003-01-01

    目的探讨溶液pH对诱变剂ICR-170诱变日本血吸虫雌虫生殖腺的影响程度.方法用5种pH值(pH 7.2-8.0)的ICR-170溶液处理日本血吸虫尾蚴30 min或45 min后,经皮肤接种小鼠观察雌虫诱变率和成虫回收率.结果溶液pH值对ICR-170的诱变作用影响较大,诱变率随着溶液pH值的上升而下降.10μg/ml 45 min组和15μg/ml 30 min组在pH 7.2时的雌虫诱变率分别为pH 7.8时的13倍和6倍,当pH>7.4时,2种浓度的ICR-170引起的诱变率基本接近.溶液pH值对于成虫回收率的影响程度远低于对诱变率的影响程度.结论在诱变日本血吸虫雌虫生殖腺时,ICR-170具有较强的pH依赖型特性.

  5. 日本血吸虫精氨酸酶基因启动子序列的扩增及序列分析%Amplification and Sequence Analysis of Schistosoma japonicum Arginase Novel Gene's Promoter Sequence

    Institute of Scientific and Technical Information of China (English)

    李孜; 余新炳; 吴忠道

    2005-01-01

    目的获取日本血吸虫(Sj)精氨酸酶(ARG)新基因的DNA编码序列及其启动子序列,实验验证ARG基因编码序列的完整性.方法 以日本血吸虫成虫 DNA做模板,PCR扩增 ARG基因编码区的DNA序列并测序;根据Sj ARG基因已扩增的 DNA序列设计2条巢式引物,用TaKaRa LA Taq PCR Cloning in vitro Kit,扩增 Sj ARG基因的启动子序列;对已扩增得到的序列进行 TATA盒的寻找以分析启动子序列的位置,实验验证我们曾获取的 Sj ARG新基因编码序列的完整性. 结果 扩增得到长约 1 000bp ARG基因 DNA序列 , Sj ARG基因 DNA序列内没有内含子,与 cDNA序列完全一致.对启动子序列扩增后,得到一略大于 250bp的序列,测序后分析发现其中有 36bp与 ARG基因 DNA序列的 5′端重叠.因此 ,将精氨酸酶基因的 DNA序列又向前延伸了 221bp,与前面得到的序列拼接后得到一条 1 486bp的总序列,将该序列在 NCBI上进行 ORF的寻找发现其最长的 ORF达 1 164bp,起始密码子在第 156位,终止密码子在第 1 319位 ,该起始密码子前 32位为 TATA盒的位置.因此,确定该 Sj ARG基因全长 cDNA编码序列长应为 1 164bp.结论 成功扩增获得了日本血吸虫精氨酸酶基因编码区的 DNA序列,其不含有内含子;并扩增得到了该基因的启动子序列,从而获得了 Sj ARG基因真正的全长 cDNA序列,为进一步的功能鉴定奠定了基础.

  6. 烟粉虱天敌日本刀角瓢虫的捕食行为%Feeding behavior of Serangium japonicum (Coleoptera: Coccinellidae), a predator of Bemisia tabaci (Homoptera: Aleyrodidae)

    Institute of Scientific and Technical Information of China (English)

    姚松林; 任顺祥; 黄振

    2005-01-01

    研究了日本刀角瓢虫雌成虫和幼虫捕食粉虱时的各种行为时间分配.结果表明,日本刀角瓢虫成虫的捕食行为可分为爬行、取食、清洁、静息、整翅和排泄6个部分.饥饿后的日本刀角瓢虫幼虫捕食烟粉虱卵时,幼虫各龄期间的取食和爬行时间无显著差异,对烟粉虱卵的处置时间随瓢虫幼虫虫龄的增大而缩短.日本刀角瓢虫雌成虫捕食烟粉虱卵时,饥饿后的雌成虫用于取食的时间显著长于非饥饿的雌成虫,而用于清洁、静息和爬行的时间显著短于非饥饿的雌成虫;饥饿后的瓢虫雌成虫对卵的处置时间显著长于非饥饿的雌成虫.饥饿后的日本刀角瓢虫雌成虫捕食烟粉虱若虫时,瓢虫雌成虫的取食时间随烟粉虱若虫虫龄的增大而增加,清洁和静息时间随烟粉虱若虫虫龄的增大而缩短,对若虫的处置时间随烟粉虱若虫虫龄的增大而增加.

  7. 曼氏和日本血吸虫副肌球蛋白的抗原肽研究%Studies on Antigenic Peptides of Paramyosins of Schistosoma mansoni and Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    杨俊海; 许家喜

    2002-01-01

    副肌球蛋白是世界卫生组织选取的用来发展血吸虫疫苗的重要候选蛋白之一.在计算机辅助下,应用PCGene程序,根据曼氏和日本血吸虫副肌球蛋白的氨基酸序列,通过对其亲水性、柔韧性、可接近性和二级结构分析预测出8个抗原肽,并用固相法进行了合成.经Dot-ELISA法测定,其中的2个对抗日本血吸虫免疫球蛋白多克隆抗体(抗-Sj-IgG-PcAb)显示出抗原性,可作为抗血吸虫病合成多肽疫苗的候选肽段.

  8. 日本血吸虫TGF-β受体I基因胞外段的克隆与表达%Cloning and expression of extracelluar fragment of TGF-β receptor Ⅰ gene of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    高志岩; 李孜; 黎小妍; 马长玲; 陈姗; 黄俊

    2011-01-01

    Objective To clone and express the extracelluar region ofTGF-β receptor-Ⅰ in Schistosomajaponicum(SjTβRIout),and to identify its immunologic functions. Methods Part of the coding region of the SjTβRI gene was obtained by rapid amplification of cDNA ends (RACE). The extracelluar fragment of SjTβRI gene was amplified by RT-PCR, cloned into the prokaryotic expression vector pET28a(+), and then confirmed by sequencing method. The plasmid was then transfected into E.coli and the expression was induced by IPTG. The recombinant protein was purified using NJ-ion affinity chromatography. SD rats were immunized with the purified protein and Freund's adjuvant. ELISA was used to detect the production of the SjTβRIout specific antibody. Results The pET28a(+)-SjTβRIout recombinant expression plasmid was successfully constructed. Sequence analysis showed that the SjTβRIout gene had 399 bp (encoding 133 amino acids) and the gene was highly homologous with Schistosoma mansoni TGF-β receptor Ⅰ. SjTβRIout recombinant protein with a molecular weight of 20 kDa was expressed in E. coli. The recombinant protein immunized SD rats produced a high titer (> 1:100 000 000) of antigen-specific IgG antibody.Conclusion The pET28a(+)-SjTβRIout recombinant protein was proved to be immunogenic.%目的 获得日本血吸虫TGF-β受体I胞外段(SjTβ3RIout)的序列,行原核克隆和表达,并分析其蛋白的免疫学功能.方法 应用cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)方法获得SjTβRI基因cDNA部分序列,用RT-PCR方法扩增SjTβRI胞外段(SjTβRlout)基因,构建pET28a(+)重组原核表达载体,并测序鉴定.将质粒转入大肠埃希菌,IPTG诱导表达,并使用Ni离子亲和层析并透析处理获得纯化重组蛋白.使用纯化蛋白与弗氏佐剂共同免疫SD大鼠获得抗血清,使用ELISA方法检测血清中SjTBRIout特异抗体IgG含量.结果 成功构建SjTβRIout原核表达载体.测序鉴定显示SjTβ3RIout包含399 bp,编码133个氨基酸,并且与曼氏血吸虫TGF-β受体I的同源性较高.重组蛋白经SDS-PAGE分析,显示其相对分子质量约为20 000,与目的 基因编码蛋白的预测分子量相符.经重组蛋白免疫的SD大鼠产生高效价(>1:100 000000)的抗原特异性IgG抗体.结论 首次获得pET28a(+)-SjTβRIout的重组蛋白,并且证明其具有良好的免疫原性.

  9. 肝片形吸虫长菲策吸虫及日本血吸虫DNA的多态性检测%Detection of DNA polymorphism among Fasciola hepatica, Fischoederius elongates and Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    胥全彬; 沈永林; 刘耀兴

    2001-01-01

    本试验用随机扩增DNA多态性技术对肝片形吸虫、长菲策吸虫和日本血吸虫虫体总DNA的多态性进行检测,结果筛选出三条具有种鉴别意义的RAPD谱带.这些特征性条带可望标记成探针用于粪便学检查中虫卵的鉴定.

  10. Dicty_cDB: CHS126 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SJS Schistosoma japonicum cDNA, mRNA sequence. 56 8e-04 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 8e-04 1 CV682854 |CV682854.1 service

  11. Dicty_cDB: SHI893 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available osoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H0...6_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G

  12. Dicty_cDB: SHD168 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  13. Dicty_cDB: SHA558 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosom...a japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sj

  14. Dicty_cDB: SHF830 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available japonicum cDNA, mRNA sequence. 56 9e-04 1 CV682907 |CV682907.1 service43_H06_sjs2...-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04 1 CV682854 |CV682854.1 service30_G09_sjs

  15. Environ: E00430 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00430 Cirsium root, rhizome or herb Crude drug Cirsium japonicum [TAX:516546], Cirsium... [TAX:41549] Asteraceae (daisy family) related species to Cirsium japonicum, Cirsium root, rhizome or he...rb (dried) Crude drugs [BR:br08305] Dicot plants: asterids Asteraceae (daisy family) E00430 Cirsium root, rhizome or herb ...

  16. Environ: E00620 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00620 Cirsium radix Crude drug Cirsium maritimum, Cirsium japonicum [TAX:516546], Cirsium suzukii, Cirsium... [TAX:41549] Asteraceae (daisy family) Cirsium maritimum, Cirsium japonicum, Cirsium... suzukii radix (dried) Crude drugs [BR:br08305] Dicot plants: asterids Asteraceae (daisy family) E00620 Cirsium radix ...

  17. Parasitosis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008013 Cloning, expression of schistosoma japonicum elastase gene and its stage-specific transcription. HUANG Chengyu(黄成玉), et al. Biotechnol Sch, East China Sci & Technol Univ, Shanghai 200237. Chin J Parasitol Parasit Dis 2007;25(5):359-363. Objective To clone, express and purify Schistosorna japonicum elastase-2b gene(SjCE-2b),

  18. Environ: E00579 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available onitum [TAX:49188] Ranunculaceae (buttercup family) Aconitum japonicum tuberous root soaked in brine and coa...t with caustic lime (dried) Crude drugs [BR:br08305] Dicot plants: others Ranunculaceae (buttercup family) E00579 Processed aconitum japonicum tuberous root ...

  19. Dicty_cDB: SHC422 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available histosoma japonicum cDNA, mRNA sequence. 56 8e-04 1 CV682907 |CV682907.1 service4...3_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 8e-04 1 CV682854 |CV682854.1 service

  20. PRODUTIVIDADE DA SOJA EM RESPOSTA Á INOCULAÇÃO PADRÃO E COINOCULAÇÃO

    Directory of Open Access Journals (Sweden)

    Ivana Marino Bárbaro

    2009-06-01

    Full Text Available The objectives of this research were to evaluate the response to a standard inoculation with Bradyrhizobium, as well as the co-inoculation with Azospirillum brasilense and Bradyrhizobium in soybean sown area already established with the culture. The rehearsal was installed in Colina-SP, in november of 2007, with the use of the soybean cultivar MG BR 46 (Conquista. The treatments were: 1 - control without N and without inoculation of Bradyrhizobium, 2-control with 200 kg N / ha split, 3-inoculation with Bradyrhizobium, 4-coinoculation with Bradyrhizobium and Azospirillum brasilense peat and 5 - co-inoculation with Bradyrhizobium and Azospirillum brasilense liquid. The experimental design was randomized blocks, with four replications. The experimental plots were constituted by six rows of four meters of length, with spacing of 0,50 m. In V6, were evaluated the nodulation, root dry mass and shoot dry mass. The crop was accomplished manually, in the useful area of the plot, being dear the yield of grains in kg/ha, after obtaining of the mass of grains for portion, being determined the text of water, calculated in 13% of humid base. Besides, the mass of the thousand seeds/plot was evaluated, in grams. In general, it was found that inoculation as well as the coinoculation did not cause an increase in most parameters assessed and soybean yield when grown in area already established with the culture, in Colina-SP.

  1. Hydroponic cultivation of soybean for Bioregenerative Life Support Systems (BLSSs)

    Science.gov (United States)

    De Pascale, Stefania; De Micco, Veronica; Aronne, Giovanna; Paradiso, Roberta

    evaluate the adaptation of soybean plants to hydroponics under controlled environment, as well as the plant response to changing cultural parameters, in order to identify the best cultivation protocol for BLSSs. The optimisation of growth conditions in hydroponics has been pursued being aware that environmental factors acting at sub-optimal levels may also increase the sensitivity of plants to space factors. The influence of the following parameters on plant growth and yield was also studied: - the hydroponic system: sole liquid solution (Nutrient Film Technique, NFT) vs solid substrate (rockwool); - the source of nitrogen in the nutrient solution: nitrate fertilizers vs urea; - the root symbiosis with atmospheric nitrogen-fixing bacteria: absence or presence of Bradyrhizobium japonicum; - the influence of microbes in the rhizosphere: inoculation with a mix containing mycorrhizal and trichoderma species, and beneficial bacteria vs a non-inoculated control. All the treatments were evaluated in terms of agronomic traits (e.g. plant size and seed production), physiological traits (gas exchange, nutrient uptake), chemical composition of seeds and their products, and technical parameters such as resource use efficiency and non-edible biomass production (waste).

  2. Whole genome co-expression analysis of soybean cytochrome P450 genes identifies nodulation-specific P450 monooxygenases

    Directory of Open Access Journals (Sweden)

    Pandey Sona

    2010-11-01

    Full Text Available Abstract Background Cytochrome P450 monooxygenases (P450s catalyze oxidation of various substrates using oxygen and NAD(PH. Plant P450s are involved in the biosynthesis of primary and secondary metabolites performing diverse biological functions. The recent availability of the soybean genome sequence allows us to identify and analyze soybean putative P450s at a genome scale. Co-expression analysis using an available soybean microarray and Illumina sequencing data provides clues for functional annotation of these enzymes. This approach is based on the assumption that genes that have similar expression patterns across a set of conditions may have a functional relationship. Results We have identified a total number of 332 full-length P450 genes and 378 pseudogenes from the soybean genome. From the full-length sequences, 195 genes belong to A-type, which could be further divided into 20 families. The remaining 137 genes belong to non-A type P450s and are classified into 28 families. A total of 178 probe sets were found to correspond to P450 genes on the Affymetrix soybean array. Out of these probe sets, 108 represented single genes. Using the 28 publicly available microarray libraries that contain organ-specific information, some tissue-specific P450s were identified. Similarly, stress responsive soybean P450s were retrieved from 99 microarray soybean libraries. We also utilized Illumina transcriptome sequencing technology to analyze the expressions of all 332 soybean P450 genes. This dataset contains total RNAs isolated from nodules, roots, root tips, leaves, flowers, green pods, apical meristem, mock-inoculated and Bradyrhizobium japonicum-infected root hair cells. The tissue-specific expression patterns of these P450 genes were analyzed and the expression of a representative set of genes were confirmed by qRT-PCR. We performed the co-expression analysis on many of the 108 P450 genes on the Affymetrix arrays. First we confirmed that CYP93C5 (an

  3. Genotypic Characterization of Bradyrhizobium Strains Nodulating Endemic Woody Legumes of the Canary Islands by PCR-Restriction Fragment Length Polymorphism Analysis of Genes Encoding 16S rRNA (16S rDNA) and 16S-23S rDNA Intergenic Spacers, Repetitive Extragenic Palindromic PCR Genomic Fingerprinting, and Partial 16S rDNA Sequencing

    OpenAIRE

    Vinuesa, Pablo; Rademaker, Jan L. W.; de Bruijn, Frans J.; Werner, Dietrich

    1998-01-01

    We present a phylogenetic analysis of nine strains of symbiotic nitrogen-fixing bacteria isolated from nodules of tagasaste (Chamaecytisus proliferus) and other endemic woody legumes of the Canary Islands, Spain. These and several reference strains were characterized genotypically at different levels of taxonomic resolution by computer-assisted analysis of 16S ribosomal DNA (rDNA) PCR-restriction fragment length polymorphisms (PCR-RFLPs), 16S-23S rDNA intergenic spacer (IGS) RFLPs, and repeti...

  4. Dicty_cDB: SHE841 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available ice43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 ser...V688215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 serv...vice30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001

  5. Dicty_cDB: SHG508 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.... 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.00...1 CV688215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001

  6. Dicty_cDB: SHB650 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04... 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-... CV688215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04

  7. Dicty_cDB: SHC807 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  8. Dicty_cDB: SHI279 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06..._sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G0

  9. Dicty_cDB: SHB339 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 9e-04 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 5...6 9e-04 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence.

  10. Dicty_cDB: SHD759 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA... sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mR

  11. Dicty_cDB: SHD160 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-1...6-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  12. Dicty_cDB: SHB464 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-...2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14

  13. Dicty_cDB: SHE173 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sj...s2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_s

  14. Dicty_cDB: SHC815 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available istosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Sc...histosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS

  15. Dicty_cDB: SHH893 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-...T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C0

  16. Dicty_cDB: SHI325 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06..._sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G0

  17. Dicty_cDB: SHD254 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  18. Dicty_cDB: SHA268 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  19. Dicty_cDB: SHK266 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available S Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 S...JS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3

  20. Dicty_cDB: SHI112 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-1...6-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-

  1. Dicty_cDB: SHI635 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available uence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA se...quence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA

  2. Dicty_cDB: SHC866 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-1...6-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-

  3. Dicty_cDB: SHH781 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available JS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 ...SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T

  4. Dicty_cDB: SHE357 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRN...A sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, m

  5. Dicty_cDB: SHJ369 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available NA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum c...DNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum

  6. Dicty_cDB: CHQ712 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma... japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistoso

  7. Dicty_cDB: SHG272 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available _sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H0...6_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G

  8. Dicty_cDB: SHB841 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 6 9e-04 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. ...56 9e-04 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA sequence

  9. Dicty_cDB: CHQ882 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available s2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_s...js2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_

  10. Dicty_cDB: SHA264 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-...2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14

  11. Dicty_cDB: SHI290 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available s2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_s...js2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_

  12. Dicty_cDB: SHL308 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available V688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...uence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mRNA s

  13. Dicty_cDB: SHI182 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available s2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_s...js2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_

  14. Dicty_cDB: SHB747 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available S Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 S...JS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3

  15. Dicty_cDB: SHF162 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available js2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_...sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09

  16. Dicty_cDB: SHE484 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  17. Dicty_cDB: SHI572 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available tosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schi...stosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Sc

  18. Dicty_cDB: CHQ520 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06..._sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G0

  19. Dicty_cDB: SHI833 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06..._sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G0

  20. Dicty_cDB: SHI645 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available _G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service4...3_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  1. Dicty_cDB: SHJ890 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs...2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sj

  2. Dicty_cDB: SHE862 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T...3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09

  3. Dicty_cDB: SHC378 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 8215 |CV688215.1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 9e-04 1 CV682854 |CV682854.1 servic

  4. Dicty_cDB: SHF776 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  5. Dicty_cDB: SHD841 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available osoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schis...tosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Sch

  6. Dicty_cDB: SHG332 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available -2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  7. Dicty_cDB: SHJ176 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available .1 sjs2-16-2_G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service...43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  8. Dicty_cDB: CHR610 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available _G01_sjs2-16-2G01-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service4...3_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service

  9. Parasitosis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008246 Effect of paeoniflorin on hepatic immunopathogenesis in mice with Schistosoma japonicum infection. CHU Deyong(储德勇),et al.Dept Parasitol,Anhui Medical Univ,Hefei 230032.Chin J Parasitol Parasit Dis 2008;26(1):10-15.Objective To investigate the mechanism of paeoniflorin in preventing hepatic granuloma formation and fibrosis in mice infected withSchistosoma japonicum.Methods Model of hepatic granuloma and fibrosis was

  10. Dicty_cDB: SHA786 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available sequence. 56 9e-04 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA, mR...NA sequence. 56 9e-04 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, m

  11. Dicty_cDB: SHC203 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available A sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schistosoma japonicum cDNA, mR...NA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schistosoma japonicum cDNA,

  12. Dicty_cDB: SHE533 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available soma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B11-T3 SJS Schist...osoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-10C09-T3 SJS Schi

  13. Dicty_cDB: SHI789 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682907 |CV682907.1 service43_H06_sjs2-16-2B...11-T3 SJS Schistosoma japonicum cDNA, mRNA sequence. 56 0.001 1 CV682854 |CV682854.1 service30_G09_sjs2-14-1

  14. Dicty_cDB: Contig-U10248-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available stosoma japonicum SJCHGC02833 ... 130 3e-34 AB241242_1( AB241242 |pid:none) Symbiotic protist of Reticuliter...FN320591 |pid:none) Schistosoma japonicum isolate Anhu... 130 7e-34 AB241243_1( AB241243 |pid:none) Symbiotic... 115 2e-32 CR932686_1( CR932686 |pid:none) Paramecium tetraurelia, Small GTPa... 124 2e-32 AB241241_1( AB241241 |pid:none) Symbiotic

  15. 核酸疫苗Sj14-3-3联合CpG和mIL-12免疫小鼠抗日本血吸虫攻击感染的研究%Immune protection of Sj14-3-3 nucleic acid vaccination with mIL-12 and CpG as adjuvants against Schistosoma japonicum in mice

    Institute of Scientific and Technical Information of China (English)

    徐元宏; 胡元生; 沈继龙

    2006-01-01

    目的在证明核酸疫苗pcDNA3.1(+)-Sj14-3-3具有部分抗血吸虫感染的基础上,联合使用CpG和pcDNA3.1(+)-mIL-12,观察此二种佐剂在攻击感染小鼠的免疫效果及其抗血吸虫感染的保护机制.方法分别用pcDNA3.1(+)-Sj14-3-3+pcDNA3.1(+)-mIL-12、pcDNA3.1(+)-Sj14-3-3+CpG、pcDNA3.1(+)-mIL-12和CpG免疫小鼠.攻击感染后6 w计数成虫负荷和肝虫卵数;检测免疫后0 w、6 w和12 w小鼠血清总IgG、IgG1和IgG2a水平;检测小鼠脾细胞培养上清中IFN-γ和IL-4;流式细胞术检测免疫鼠脾细胞中CD4+和CD8+T细胞的比率.结果pcDNA3.1(+)-Sj14-3-3+pcDNA3.1(+)-mIL-12和pcDNA3.1(+)-Sj14-3-3+CpG免疫小鼠的减虫率分别为41.2%和28.7%;减卵率分别为52.6%和41.2%.单独使用pcDNA3.1(+)-mIL-12和CpG也有一定的免疫保护作用.保护性免疫主要通过诱导宿主产生CTL、TH1型和体液免疫应答.结论pcDNA3.1(+)-mIL-12和CpG具有较强的增强核酸疫苗pcDNA3.1(+)-Sj14-3-3抗血吸虫攻击感染作用.

  16. CLONING AND FUNCTION ANALYSIS OF THE FULL LENGTH SEQUENCE OF A NEW SCHISTOSOMA JAPONICUM ADENYLATE KINASE%日本血吸虫一种新腺苷酸激酶全长cDNA的克隆与功能分析

    Institute of Scientific and Technical Information of China (English)

    彭鸿娟; 陈晓光; 王珣章

    2002-01-01

    目的对用表达序列标签(Expression Sequence Tag,EST)策略从日本血吸虫尾蚴cDNA文库中筛选出的新基因进行功能预测.方法用NCBI站点的BLASTx及BLASTn程序对所获得的新基因序列进行同源性搜索;用NCBI BLAST站点的blast two sequence程序对同源性高的基因进行核苷酸及氨基酸水平的同源性比较;利用Motif Scan in a Protein Sequence对cDNA序列所编码的蛋白质进行结构域搜索;利用CD-Search程序对目标蛋白进行保守区域搜索.结果发现一个与曼氏血吸虫22 kDa单核苷酸激酶mRNA高度同源的日本血吸虫新基因,基因与氨基酸水平的同一性均分别为86.0%和87.0%;蛋白结构域搜索及保守区域搜索结果显示,该cDNA序列所编码的蛋白质是一种腺苷酸激酶. 结论用EST策略筛选新基因是一个可行的方法,所筛到的日本血吸虫新基因编码一种腺苷酸激酶,全长编码序列与曼氏血吸虫腺苷酸激酶mRNA高度同源.

  17. 2010年全国重点水域血吸虫感染哨鼠监测预警情况分析%Surveillance and forecast of Schistosoma japonicum-infected sentinel mice in key water regions of China in 2010

    Institute of Scientific and Technical Information of China (English)

    郑浩; 杨坤; 李石柱; 许静; 郭家钢; 陈朝; 周晓农; 孙乐平; 朱蓉; 涂祖武; 李以义; 杨卫平; 辜小南; 吴子松; 冯锡光

    2012-01-01

    目的 探索血吸虫病监测预警的方法与手段,为提高血吸虫病监测预警系统的敏感性提供科学依据.方法 在湖南、湖北、江西、安徽、江苏、云南和四川7个省选择血吸虫病易感重点水域,采用哨鼠尾蚴测定法,分别于2010年6月和9月开展2次现场检测.哨鼠在实验室饲养后解剖观察血吸虫感染情况,建立全国重点水域血吸虫感染性数据库,分析哨鼠监测预警阳性点的时空分布和环境特点等情况.结果 2010年6月和9月在7个省34个县(市、区)检测了72个点.共投放哨鼠2 667只,回收哨鼠2613只,总回收率为97.98%;共检出17个阳性点,阳性点出现率为23.61%,其中6月份阳性点出现率为17.24%(10/58),9月份阳性点出现率为14.71%( 10/68),差异无统计学意义(x2=0.151,P=0.698).共解剖哨鼠2436只,检出阳性哨鼠90只,检获血吸虫459条,哨鼠总感染率为3.69%,阳性鼠平均虫负荷为5.10条/鼠.6月份哨鼠感染率为2.82%(31/1 099),阳性鼠平均虫负荷为2.45条/鼠;9月份哨鼠感染率为4.41%(59/1 337),阳性鼠平均虫负荷为6.49条/鼠,9月份哨鼠感染率显著高于6月份(x2=14.681,P<0.01).当年有感染性钉螺、上年有感染性钉螺和近3年无感染性钉螺环境哨鼠阳性点检出率分别为29.63%、41.67%和12.12%,差异无统计学意义(x2=5.227,P=0.071);上述3类环境哨鼠感染率分别为9.38%、3.98%和0.59%,差异有统计学意义(x2=20.489,P< 0.01).结论 重点水域哨鼠感染监测结果能基本反映当地血吸虫病疫情,对近年未查出感染性钉螺的环境仍能检出较多阳性.采用哨鼠测定法在重点水域进行监测预警将显著提高血吸虫病监测预警系统的敏感性.%Objective To explore the approach and tool for surveillance and forecast of schistosomiasis, so as to provide sci-entific evidence for improving the sensitivity of the surveillance and forecast system of schistosomiasis. Methods Two field detections were carried out in June and September by using the determination of sentinel mice in key high-risk water regions of 7 provinces of Hunan, Hubei, Jiangxi, Anhui, Jiangsu, Yunnan and Sichuan, and the sentinel mice were raised in laboratory and dissected for observation of schistosome infections. The database regarding schisosome infections in key water regions of China was established, and the tempo-spatial distribution and environmental features of the national surveillance and forecast sites with positive sentinel mice were analyzed. Results A total of 72 surveillance and forecast sites were detected in 34 counties (cities, districts) of 7 provinces. Of the 2 667 sentinel mice placed, 2 613 were recovered, with a recovery rate of 97.98%. Among the 72 sites detected, 17 were detected with positive sentinel mice, and the occurrence rate of positive sites was 23.61%. The occurrence rate was 17.24% (10/58) in June and 14.71% (10/68) in September, and no significant differ-ence was observed between two batches ( x2 = 0.151, P - 0.698). Of the 2 436 sentinel mice dissected, 90 were positive, and 459 schistosome worms were collected, with a total infection rate of 3.69%, and the mean worm burdens of positive mice were 5.10 worms per mouse. The infection rate of sentinel mice was 2.82% (31/1 099) in June and 4.41% (58/1 337) in Sep-tember, the infection rate of sentinel mice was significantly higher in September than that in Juen ( x2 = 14.681, P < 0.01) , and the mean worm burdens of infected sentinel mice were 2.45 worms per mouse in June and 6.49 worms per mouse in Sep-tember. The occurrence rates of the positive sites with infected snails detected in the study year, last year and without infected snails detected in recent 3 years were 29.63%, 41.67% and 12.12%, respectively, and no significant differences were detect-ed ( x2 = 5-227, P = 0.071). The infection rates of sentinel mice in the aforementioned 3 settings were 9.38%, 3.98% and 0.59%, respectively, and there was significant difference observed ( x2 = 20.489, P < 0.01). Conclusions The monitoring results of sentinel mice infections in key water regions can almost reflect the endemic situation of schistosomiasis in local ar-eas. Notably, many infections are detected in settings without infected snails detected in recent years. The monitoring of senti-nel mice infections could significantly improve the sensitivity of the surveillance and forecast system of schistosomiasis.

  18. Investigation on prevalence and risk factors of HIV/AIDS and Schistosoma japonicum, Ascaris lumbricoides and Trichuris trichiura infections in a rural community of southwestern China%我国西南彝族农村地区HIV与血吸虫蛔虫及鞭虫感染现状调查

    Institute of Scientific and Technical Information of China (English)

    肖鹏磊; 周艺彪; 史妍; 杨亚; 高建川; 宋秀霞; 姜庆五

    2016-01-01

    目的 了解我国西南彝族农村地区HIV、血吸虫、蛔虫及鞭虫感染状况,调查是否存在混合感染.方法 在我国西南部某山区随机选取一个彝族乡开展横断面调查,检测村民HIV、血吸虫、蛔虫及鞭虫感染情况;并对村民进行问卷调查,了解HIV及寄生虫感染的危险因素.结果 我国西南彝族农村地区居民HIV、血吸虫、蛔虫和鞭虫感染率分别为2.33% 、2.05%、13.47%和30.59%,鞭虫与蛔虫共同感染检出率为7.08%,HIV与蛔虫和鞭虫共同感染检出率均为0.23%.男性以及吸毒是HIV感染的危险因素[OR=3.26,95% CI:(0.97,10.95);OR=72.86,95% CI:(18.51,286.76)];厕所与蛔虫和鞭虫感染间均存在负相关关系[OR=0.51,95% CI:(0.27,0.98);OR=0.48,95% CI:(0.28,0.80)].与居住在村4的居民相比,村1村民感染蛔虫风险较高[OR=3.14,95% CI:(1.35,7.27)];与居住在村4的居民相比,居住在村2和村3的村民鞭虫感染风险较高[OR=3.73,95% CI:(1.92,7.26);OR=4.53,95% CI:(2.12,9.68)].此外,11~ 20岁年龄组村民鞭虫感染风险高于>50岁村民[OR=3.72,95% CI:(1.59,8.67)];蛔虫与鞭虫感染间存在相关关系[OR=3.11,95% CI:(1.63,5.93)],未发现与血吸虫感染相关的因素.结论 我国西南彝族农村居民HIV及寄生虫感染率尤其是鞭虫感染率较高;应进一步探索HIV与寄生虫感染,尤其是与血吸虫感染间的关系.

  19. Effect of artemether combming with heme on activities of phosphogly cerate kinase and pyruvate kinase in adult Schistosoma japonicum in vitro%体外蒿甲醚伍用血红素对血吸虫磷酸甘油酸激酶和丙酮酸激酶的影响

    Institute of Scientific and Technical Information of China (English)

    翟自立; 焦佩英; 梅静艳; 郭惠芳; 肖树华

    2002-01-01

    目的观察体外蒿甲醚与血红素伍用对日本血吸虫磷酸甘油酸激酶(PGK)和丙酮酸激酶(PK)的影响.方法以4~5周龄的血吸虫,置于含蒿甲醚和/或血红素的培素液内培养24h后,测定虫体的PGK和PK酶活力.结果体外,蒿甲醚(50μmo1/L)或血红素(50μmo1/L)对血吸虫的PGK和PK酶活力无影响,但两者伍用可引起两种酶的活力明显下降.结论蒿甲醚对血吸虫PGK和PK的影响具有血红素依赖性.

  20. BIOCHEMICAL AND IMMUNOLOGICAL CHARACTERIZATION OF THE 200kD SCHISTOSOMULUM SURFACE ANTIGEN COMMON TO SCHISTOSOMA MANSONI AND SCHISTOSOMA JAPONICUM%曼氏和日本血吸虫童虫体表200kD共同抗原的生化与免疫学特性

    Institute of Scientific and Technical Information of China (English)

    易新元; 周金春; 王庆林

    2000-01-01

    Objective To study the biochemecal and immunological characterization of the 200 kD schistosomulum surface antigen Method and results A very high molecular weight schistosomulum surface antigen of Mr>200kD was identified and characterized using monoclonal antibodies. Carbohydrate modification experiments followed by radioimmunobinding assays demonstrated that the epitope recognised by the mAbs on this antigen was carbohydrate in nature, while protein digestion experiments followed by SDS-PAGE indicated that this antigen also contained protein. Immunoprecipitation of 125I-labelled cercarial, schistosomulum, adult worm and miracidial surface antigens followed by gel analysis showed the carbohydrate epitope to be present on 5 cercarial, 2 schistosomulum and 5 miracidial surface molecules, and suggested a possible ecological function involved in adapting the parasite to the aquatic free-living stages of its life cycle and possibly also in protecting the early schistosomula from host immune damage. The 5 cercarial surfacs antigens proved to be associated with the CHR, since all the mAbs which recognised those antigens could induce a strong CHR. A kinetic investigation of the carbohydrate epitope on schistosomula of different ages demonstrated a gradual and possibly irreversible loss during the culture period. The epitope completely disappeared from the surface of adult worms. Conclusion To demonstrate an epitope common to a number of surface molecules of various developmental stages of schistosome and therefore explains the immunological cross - reactivity among different stages at the molecular level.