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Sample records for bradyrhizobium japonicum infection1cwoa

  1. Molybdate transport by Bradyrhizobium japonicum bacteroids.

    OpenAIRE

    Maier, R J; Graham, L.

    1988-01-01

    Bacteroid suspensions of Bradyrhizobium japonicum USDA 136 isolated from soybeans grown in Mo-deficient conditions were able to transport molybdate at a nearly constant rate for up to 1 min. The apparent Km for molybdate was 0.1 microM, and the Vmax was about 5 pmol/min per mg (dry weight) of bacteroid. Supplementation of bacteroid suspensions with oxidizable carbon sources did not markedly increase molybdate uptake rates. Anaerobically isolated bacteroids accumulated twice as much Mo in 1 h ...

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

    Science.gov (United States)

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

    2014-04-01

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

  3. Symbiotic efficiency of selected indigenous Bradyrhizobium japonicum strains

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    Branka Komesarović

    2007-12-01

    Full Text Available Symbiotic nitrogen fixation has a unique role in a sustainable agriculture because the utilization of this natural process enables significant reduction of mineral nitrogen fertilization. Thereby, selection of highly efficient symbiotic nitrogen fixators is of great importance for successful legume inoculation due to the fact that the strains of nodule bacteria strongly differ in their symbiotic efficiency, competitiveness and compatibility. The main aim of the present study was to evaluate the symbiotic characteristics of indigenous and reference Bradyrhizobium japonicum and Bradyrhizobium elkanii strains and their compatibility with different soybean cultivars in agroecological conditions of eastern Slavonia. During two years of investigations, twofactorial field trials were set up in a randomized complete block design with five replicates. The results obtained in this study show that all B. japonicum strains had positive effect on all analyzed properties and particularly on seed yield as the most important parameter in soybean production. The highest values for nodule dry weight were obtained by application of indigenous B.japonicum strain S69 that was isolated from the similar agroecologicalconditions. Although the application of B. elkanii strain S76T resulted in the most abundant nodulation, at the same time it induced chlorosis on leaves in full blossom stage. Regarding the tested soybean cultivars, Ika and Nada, significant differences in nodulation were determined. Considerable rainfall deficiency in 2002 had strong influence on all tested properties. The highest nitrogen content in soybean aerial parts as well as the seed yield were determined in application of indigenous B. japonicum strains S69 and S75. The obtained results suggest that these two indigenous strains can becharacterized as the most efficient ones and could be used for soybean inoculant production for that region.

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

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    Julieta Pérez-Giménez

    2009-01-01

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

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

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    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.

  6. Influence of oxygen on DNA binding, positive control, and stability of the Bradyrhizobium japonicum NifA regulatory protein.

    OpenAIRE

    1991-01-01

    Central to the genetic regulatory circuit that controls Bradyrhizobium japonicum nif and fix gene expression is the NifA protein. NifA activates transcription of several nif and fix genes and autoregulates its expression during symbiosis in soybean root nodules or in free-living microaerobic conditions. High O2 tensions result in the lack of nif expression, possibly by inactivation of NifA through oxidation of an essential metal cofactor. Several B. japonicum nif and fix promoters have upstre...

  7. Characterization of a Functional Role of the Bradyrhizobium japonicum Isocitrate Lyase in Desiccation Tolerance

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    Jeong-Min Jeon

    2015-07-01

    Full Text Available Bradyrhizobium japonicum is a nitrogen-fixing symbiont of soybean. In previous studies, transcriptomic profiling of B. japonicum USDA110, grown under various environmental conditions, revealed the highly induced gene aceA, encoding isocitrate lyase (ICL. The ICL catalyzes the conversion of isocitrate to succinate and glyoxylate in the glyoxylate bypass of the TCA cycle. Here, we evaluated the functional role of B. japonicum ICL under desiccation-induced stress conditions. We purified AceA (molecular mass = 65 kDa from B. japonicum USDA110, using a His-tag and Ni-NTA column approach, and confirmed its ICL enzyme activity. The aceA mutant showed higher sensitivity to desiccation stress (27% relative humidity (RH, compared to the wild type. ICL activity of the wild type strain increased approximately 2.5-fold upon exposure to 27% RH for 24 h. The aceA mutant also showed an increased susceptibility to salt stress. Gene expression analysis of aceA using qRT-PCR revealed a 148-fold induction by desiccation, while other genes involved in the glyoxylate pathway were not differentially expressed in this condition. Transcriptome analyses revealed that stress-related genes, such as chaperones, were upregulated in the wild-type under desiccating conditions, even though fold induction was not dramatic (ca. 1.5–2.5-fold.

  8. Survival and Competitiveness of Bradyrhizobium japonicum Strains 20 Years after Introduction into Field Locations in Poland.

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    Narożna, Dorota; Pudełko, Krzysztof; Króliczak, Joanna; Golińska, Barbara; Sugawara, Masayuki; Mądrzak, Cezary J; Sadowsky, Michael J

    2015-08-15

    It was previously demonstrated that there are no indigenous strains of Bradyrhizobium japonicum forming nitrogen-fixing root nodule symbioses with soybean plants in arable field soils in Poland. However, bacteria currently classified within this species are present (together with Bradyrhizobium canariense) as indigenous populations of strains specific for nodulation of legumes in the Genisteae tribe. These rhizobia, infecting legumes such as lupins, are well established in Polish soils. The studies described here were based on soybean nodulation field experiments, established at the Poznań University of Life Sciences Experiment Station in Gorzyń, Poland, and initiated in the spring of 1994. Long-term research was then conducted in order to study the relation between B. japonicum USDA 110 and USDA 123, introduced together into the same location, where no soybean rhizobia were earlier detected, and nodulation and competitive success were followed over time. Here we report the extra-long-term saprophytic survival of B. japonicum strains nodulating soybeans that were introduced as inoculants 20 years earlier and where soybeans were not grown for the next 17 years. The strains remained viable and symbiotically competent, and molecular and immunochemical methods showed that the strains were undistinguishable from the original inoculum strains USDA 110 and USDA 123. We also show that the strains had balanced numbers and their mobility in soil was low. To our knowledge, this is the first report showing the extra-long-term persistence of soybean-nodulating strains introduced into Polish soils and the first analyzing the long-term competitive relations of USDA 110 and USDA 123 after the two strains, neither of which was native, were introduced into the environment almost 2 decades ago.

  9. An iron uptake operon required for proper nodule development in the Bradyrhizobium japonicum-soybean symbiosis.

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    Benson, Heather P; Boncompagni, Eric; Guerinot, Mary Lou

    2005-09-01

    Rhizobia live in the soil or enter into a nitrogen-fixing symbiosis with a suitable host plant. Each environment presents different challenges with respect to iron acquisition. The soybean symbiont Bradyrhizobium japonicum 61A152 can utilize a variety of siderophores (Fe[III]-specific ligands). Purification of iron-regulated outer membrane proteins had previously allowed the cloning of a gene, fegA, from B. japonicum 61A152, whose predicted protein shares significant amino acid similarity with known TonB-dependent siderophore receptors. Here, we show that fegA is in an operon with a gene, fegB, that is predicted to encode an inner membrane protein. Characterization of fegAB and fegB mutants shows that bothfegA and fegB are required for utilization of the siderophore ferrichrome. Whereas thefegB mutant forms a normal symbiosis, the fegAB mutant has a dramatic phenotype in planta. Six weeks after inoculation with a fegAB strain, soybean nodules do not contain leghemoglobin and do not fix nitrogen. Infected cells contain few symbiosomes and are filled with vesicles. As ferrichrome is a fungal siderophore not likely to be available in nodules, the symbiotic defect suggests that the fegAB operon is serving a different function in planta, possibly one involved in signaling between the two partners.

  10. (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.

  11. [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.

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

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    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.

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

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    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.

  14. 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

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    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

  15. 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.

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

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    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.

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

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese R.; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Paša-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-01-01

    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 eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 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. PMID:22843990

  18. Quantitative phosphoproteomic analysis of soybean root hairs inoculated with Bradyrhizobium japonicum.

    Science.gov (United States)

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

    2012-11-01

    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 eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 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.

  19. 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.

  20. 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.

  1. 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

  2. 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

    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 consistin...

  3. 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.

  4. Interspecies Complementation of Escherichia coli ccm Mutants: CcmE (CycJ) from Bradyrhizobium japonicum Acts as a Heme Chaperone during Cytochrome c Maturation

    OpenAIRE

    Schulz, Henk; Thöny-Meyer, Linda

    2000-01-01

    Biogenesis of c-type cytochromes in α- and γ-proteobacteria requires the function of a set of orthologous genes (ccm genes) that encode specific maturation factors. The Escherichia coli CcmE protein is a periplasmic heme chaperone. The membrane protein CcmC is required for loading CcmE with heme. By expressing CcmE (CycJ) from Bradyrhizobium japonicum in E. coli we demonstrated that heme is bound covalently to this protein at a strictly conserved histidine residue. The B. japonicum homologue ...

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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).

  10. 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.

  11. The major Nod factor of Bradyrhizobium japonicum promotes early growth of soybean and corn.

    Science.gov (United States)

    Souleimanov, A; Prithiviraj, B; Smith, D L

    2002-09-01

    Greenhouse experiments were conducted to evaluate the effect of Nod factor Nod Bj-V (C18:1, MeFuc) of Badyrhizobium japonicum on the growth of soybean and corn. Three-day-old seedlings of soybean and corn were grown in hydroponic solutions containing four concentrations (0, 10(-7), 10(-9) or 10(-11) M) of Nod factor. After 7 d of treatment, Nod factor enhanced soybean and corn biomass. Nod factor elicited profound effects on root growth resulting in 34-44% longer roots in soybean. More detailed analyses of the roots, using a scanner based image analysis system, revealed that Nod factor increased the total length, projected area and surface area of the roots and decreased the diameter of soybean roots, while it increased the total length of corn roots. Stem injection of soybean plants with 10(-7) M Nod factor resulted in increased dry matter accumulation. These results suggest that Nod factor, besides mediating early stages of nodulation, has more general plant growth-promoting effects.

  12. The Bradyrhizobium japonicum fegA gene encodes an iron-regulated outer membrane protein with similarity to hydroxamate-type siderophore receptors.

    Science.gov (United States)

    LeVier, K; Guerinot, M L

    1996-12-01

    Iron is important in the symbiosis between soybean and its nitrogen-fixing endosymbiont Bradyrhizobium japonicum, yet little is known about rhizobial iron acquisition strategies. Analysis of outer membrane proteins (OMPs) from B. japonicum 61A152 identified three iron-regulated OMPs in the size range of several known receptors for Fe(III)-scavenging siderophores. One of the iron-regulated proteins, FegA, was purified and microsequenced, and a reverse genetics approach was used to clone a fegA-containing DNA fragment. Sequencing of this fragment revealed a single open reading frame of 750 amino acids. A putative N-terminal signal sequence of 14 amino acids which would result in a mature protein of 736 amino acids with a molecular mass of 80,851 Da was predicted. FegA shares significant amino acid similarity with several Fe(III)-siderophore receptors from gram-negative bacteria and has greater than 50% amino acid similarity and 33% amino acid identity with two [corrected] bacterial receptors for hydroxamate-type Fe(III)-siderophores. A dendrogram describing total inferred sequence similarity among 36 TonB-dependent OMPs was constructed; FegA grouped with Fe(III)-hydroxamate receptors. The transcriptional start site of fegA was mapped by primer extension analysis, and a putative Fur-binding site was found in the promoter. Primer extension and RNA slot blot analysis demonstrated that fegA was expressed only in cells grown under iron-limiting conditions. This is the first report of the cloning of a gene encoding a putative Fe(III)-siderophore receptor from nitrogen-fixing rhizobia.

  13. 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.

  14. 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.

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    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.

  15. 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.

  16. Inoculation with an enhanced N2 -fixing Bradyrhizobium japonicum strain (USDA110) does not alter soybean (Glycine max Merr.) response to elevated [CO2 ].

    Science.gov (United States)

    Sanz-Sáez, Álvaro; Heath, Katy D; Burke, Patricia V; Ainsworth, Elizabeth A

    2015-12-01

    This study tested the hypothesis that inoculation of soybean (Glycine max Merr.) with a Bradyrhizobium japonicum strain (USDA110) with greater N2 fixation rates would enhance soybean response to elevated [CO2 ]. In field experiments at the Soybean Free Air CO2 Enrichment facility, inoculation of soybean with USDA110 increased nodule occupancy from 5% in native soil to 54% in elevated [CO2 ] and 34% at ambient [CO2 ]. Despite this success, inoculation with USDA110 did not result in greater photosynthesis, growth or seed yield at ambient or elevated [CO2 ] in the field, presumably due to competition from native rhizobia. In a growth chamber experiment designed to study the effects of inoculation in the absence of competition, inoculation with USDA110 in sterilized soil resulted in nodule occupation of >90%, significantly greater (15) N2 fixation, photosynthetic capacity, leaf N and total plant biomass compared with plants grown with native soil bacteria. However, there was no interaction of rhizobium fertilization with elevated [CO2 ]; inoculation with USDA110 was equally beneficial at ambient and elevated [CO2 ]. These results suggest that selected rhizobia could potentially stimulate soybean yield in soils with little or no history of prior soybean production, but that better quality rhizobia do not enhance soybean responses to elevated [CO2 ].

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. 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 %.

  2. 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.

  3. 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.

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

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    Dušica Delić

    2010-04-01

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

  5. 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.

  6. 菌根与根瘤菌联合应用对复垦矿区根际土壤环境的改良后效%Lasting improvement effects of arbuscular mycorrhizal fungi and Bradyrhizobium japonicum on rhizosphere soil environment in mining subsidence

    Institute of Scientific and Technical Information of China (English)

    于淼; 毕银丽; 张翠青

    2013-01-01

    In a mining subsidence experiment in the Shendong mining area in the Shaanxi province of China, agricultural soil was improved by inoculation with arbuscular mycorrhizal fungi and Bradyrhizobium japonicum. This experiment was implemented in mid-May 2008 for the first planting, and in the end of April 2009 for the second time. It was divided into two experiment plots, inoculated and control. After two years of abandoned cropland, in September 2011, 15 random positions were selected under the"S"shape method from each district, and roots and fresh rhizosphere soil samples were collected. The aftereffect of microbial reclamation of the soil was studied systematically, about which there have been no reports until now. The results were 1) the density of external hyphal of the inoculation treatment was 5.58 m/g, 90%higher than the control after abandonment, the difference reaching a significant level, and it was also significantly higher than the two sets of processing before abandonment;2) the root infection rate of the inoculation treatment was 91%, 52%higher than the control after abandonment, the difference reaching a significant level, and it was significantly higher than the control before abandonment; 3) pH value of the inoculation treatment was significantly higher than that of the control after abandonment;4) in the inoculation treatment the number of bacteria, microbial total, available phosphorus content, conductivity and acid phosphatase activity were also greater than that of the control after abandonment, the differences not reaching a significant level; 5)compared with before and after abandonment, the soil was in degraded trends;in a comparison between the years of 2009 and 2011, in the control treatment there was a slight increase in hyphal density, the difference not reaching a significant level, while in the inoculation treatment hyphal density was twice than that before abandonment, the difference reaching a significant level;6)compared with before and

  7. 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.

  8. 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.

  9. 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.

  10. Revision of the taxonomic status of the species Rhizobium lupini and reclassification as Bradyrhizobium lupini comb. nov.

    Science.gov (United States)

    Peix, Alvaro; Ramírez-Bahena, Martha Helena; Flores-Félix, José David; Alonso de la Vega, Pablo; Rivas, Raúl; Mateos, Pedro F; Igual, José M; Martínez-Molina, Eustoquio; Trujillo, Martha E; Velázquez, Encarna

    2015-04-01

    The species Rhizobium lupini was isolated from Lupinus nodules and included in the Approved Lists of Bacterial Names in 1980. Nevertheless, on the basis of the analysis of the type strain of this species available in DSMZ, DSM 30140(T), whose 16S rRNA gene was identical to that of the type strain of Bradyrhizobium japonicum , R. lupini was considered a later synonym of this species. In this study we confirmed that the strain DSM 30140(T) belongs to the species B. japonicum , but also that it cannot be the original strain of R. lupini because this species effectively nodulated Lupinus whereas strain DSM 30140(T) was able to nodulate soybean but not Lupinus. Since the original type strain of R. lupini was deposited into the USDA collection by L. W. Erdman under the accession number USDA 3051(T) we analysed the taxonomic status of this strain showing that although it belongs to the genus Bradyrhizobium instead of genus Rhizobium , it is phylogenetically distant from B. japonicum and closely related to Bradyrhizobium canariense . The type strains R. lupini USDA 3051(T) and B. canariense BTA-1(T) share 16S rRNA, recA and glnII gene sequences with similarities of 99.8%, 96.5% and 97.1%, respectively. They presented a DNA-DNA hybridization value of 36% and also differed in phenotypic characteristics and slightly in the proportions of some fatty acids. Therefore we propose the reclassification of the species Rhizobium lupini as Bradyrhizobium lupini comb. nov. The type strain is USDA 3051(T) ( = CECT 8630(T) = LMG 28514(T)).

  11. Development and application of a multilocus sequence analysis method for the identification of genotypes within genus Bradyrhizobium and for establishing nodule occupancy of soybean (Glycine max L. Merr).

    Science.gov (United States)

    van Berkum, Peter; Elia, Patrick; Song, Qijian; Eardly, Bertrand D

    2012-03-01

    A multilocus sequence typing (MLST) method based on allelic variation of seven chromosomal loci was developed for characterizing genotypes (GT) within the genus Bradyrhizobium. With the method, 29 distinct multilocus GT were identified among 190 culture collection soybean strains. The occupancy of 347 nodules taken from uninoculated field-grown soybean plants also was determined. The bacteroid GT were either the same as or were closely related to GT identified among strains in the culture collection. Double-nodule occupancy estimates of 2.9% were much lower than values published based on serology. Of the 347 nodules examined, 337 and 10 were occupied by Bradyrhizobium japonicum and B. elkanii, respectively. The collection strains within the species B. japonicum and B. elkaniialso were compared with Bradyrhizobium cultures from other legumes. In many cases, the observed GT varied more according to their geographic origin than by their trap hosts of isolation. In other cases, there were no apparent relationships with either the legume or geographic source. The MLST method that was developed should be a useful tool in determining the influence of geographic location, temperature, season, soil type, and host plant cultivar on the distribution of GT of Bradyrhizobium spp.

  12. Taxonomy Icon Data: Schistosoma japonicum [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available .png Schistosoma_japonicum_NL.png Schistosoma_japonicum_S.png Schistosoma_japonicum_NS.png http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Schistosoma+japonicum&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=S...chistosoma+japonicum&t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schist...osoma+japonicum&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Schistosoma+japonicum&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=132 ...

  13. Multilocus Sequence Analysis for Assessment of the Biogeography and Evolutionary Genetics of Four Bradyrhizobium Species That Nodulate Soybeans on the Asiatic Continent ▿ †

    Science.gov (United States)

    Vinuesa, Pablo; Rojas-Jiménez, Keilor; Contreras-Moreira, Bruno; Mahna, Suresh K.; Prasad, Braj Nandan; Moe, Hla; Selvaraju, Suresh Babu; Thierfelder, Heidemarie; Werner, Dietrich

    2008-01-01

    A highly supported maximum-likelihood species phylogeny for the genus Bradyrhizobium was inferred from a supermatrix obtained from the concatenation of partial atpD, recA, glnII, and rpoB sequences corresponding to 33 reference strains and 76 bradyrhizobia isolated from the nodules of Glycine max (soybean) trap plants inoculated with soil samples from Myanmar, India, Nepal, and Vietnam. The power of the multigene approach using multiple strains per species was evaluated in terms of overall tree resolution and phylogenetic congruence, representing a practical and portable option for bacterial molecular systematics. Potential pitfalls of the approach are highlighted. Seventy-five of the isolates could be classified as B. japonicum type Ia (USDA110/USDA122-like), B. liaoningense, B. yuanmingense, or B. elkanii, whereas one represented a novel Bradyrhizobium lineage. Most Nepalese B. japonicum Ia isolates belong to a highly epidemic clone closely related to strain USDA110. Significant phylogenetic evidence against the monophyly of the of B. japonicum I and Ia lineages was found. Analysis of their DNA polymorphisms revealed high population distances, significant genetic differentiation, and contrasting population genetic structures, suggesting that the strains in the Ia lineage are misclassified as B. japonicum. The DNA polymorphism patterns of all species conformed to the expectations of the neutral mutation and population equilibrium models and, excluding the B. japonicum Ia lineage, were consistent with intermediate recombination levels. All species displayed epidemic clones and had broad geographic and environmental distribution ranges, as revealed by mapping climate types and geographic origins of the isolates on the species tree. PMID:18791003

  14. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2008-09-15

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

  16. Ethyl tert-butyl ether (ETBE) biodegradation by a syntrophic association of Rhodococcus sp. IFP 2042 and Bradyrhizobium sp. IFP 2049 isolated from a polluted aquifer.

    Science.gov (United States)

    Le Digabel, Yoann; Demanèche, Sandrine; Benoit, Yves; Vogel, Timothy M; Fayolle-Guichard, Françoise

    2013-12-01

    Ethyl tert-butyl ether (ETBE) enrichment was obtained by adding contaminated groundwater to a mineral medium containing ETBE as the sole carbon and energy source. ETBE was completely degraded to biomass and CO2 with a transient production of tert-butanol (TBA) and a final biomass yield of 0.37 ± 0.08 mg biomass (dry weight).mg(-1) ETBE. Two bacterial strains, IFP 2042 and IFP 2049, were isolated from the enrichment, and their 16S rRNA genes (rrs) were similar to Rhodococcus sp. (99 % similarity to Rhodococcus erythropolis) and Bradyrhizobium sp. (99 % similarity to Bradyrhizobium japonicum), respectively. Rhodococcus sp. IFP 2042 degraded ETBE to TBA, and Bradyrhizobium sp. IFP 2049 degraded TBA to biomass and CO2. A mixed culture of IFP 2042 and IFP 2049 degraded ETBE to CO2 with a biomass yield similar to the original ETBE enrichment (0.31 ± 0.02 mg biomass.mg(-1) ETBE). Among the genes previously described to be involved in ETBE, MTBE, and TBA degradation, only alkB was detected in Rhodococcus sp. IFP 2042 by PCR, and none were detected in Bradyrhizobium sp. IFP 2049.

  17. Componentes de produção e produtividade da cultura da soja submetida à inoculação de Bradyrhizobium e Azospirillum

    Directory of Open Access Journals (Sweden)

    Lucas Guilherme Bulegon

    2016-04-01

    Full Text Available O presente trabalho teve por objetivo avaliar os componentes de produção e produtividade de dois genótipos de soja cultivados em casa de vegetação e submetidos à inoculação com bactérias diazotróficas do gênero Bradyrhizobium e Azospirillum. Adotou-se delineamento em blocos ao acaso com quatro repetições, em esquema fatorial 2 × 4; no fator 1 foram testados: dois genótipos de soja (BMX Turbo e Coodetec 250 e no fator 2 foram avaliadas quatro combinações de inoculação (sementes não inoculadas mas que receberam 200 kg ha-1 de N na semeadura; sementes inoculado com Bradyrhizobium japonicum; sementes inoculadas com Azospirillum brasilense; sementes com inoculação de B. japonicum e A. brasilense. No estádio R8 foi avaliado as variáveis biométricas: diâmetro de coleto (DC; altura de planta (ALP; número de vagens (NVS; número de grãos por vagem (NGV; massa seca de parte aérea (MPA;massa seca de vagens (MSV; produtividade por planta (PPP; teor de N na parte aérea (TNA e teor de N nos grãos (TNG. Foi observado resposta diferenciada dos genótipos de soja BMX Turbo e Coodetec 250 à inoculação de sementes com bactérias diazotróficas do gênero B. japonicum ou A. brasilense na altura de plantas e nitrogênio na parte aérea. A inoculação conjunta de B. japonicum e A. brasilense em sementes intensifica o desempenho produtivo da cultura da soja no genótipo BMX Turbo.

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

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    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

  19. 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

  20. 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

    addition of the substrates for induction and is oxygen dependent. The highest activity is obtained when the concentration of inducer is 0.2 mM. Spectrophotometric data are consistent with the suggestion that the indole ring is broken during degradation of IAA. We hypothesize that the enzyme catalyzes...... 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...

  1. 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.

  2. Bradyrhizobium pachyrhizi sp. nov. and Bradyrhizobium jicamae sp. nov., isolated from effective nodules of Pachyrhizus erosus.

    Science.gov (United States)

    Ramírez-Bahena, Martha Helena; Peix, Alvaro; Rivas, Raúl; Camacho, María; Rodríguez-Navarro, Dulce N; Mateos, Pedro F; Martínez-Molina, Eustoquio; Willems, Anne; Velázquez, Encarna

    2009-08-01

    Several strains isolated from the legume Pachyrhizus erosus were characterized on the basis of diverse genetic, phenotypic and symbiotic approaches. These novel strains formed two groups closely related to Bradyrhizobium elkanii according to their 16S rRNA gene sequences. Strains PAC48T and PAC68T, designated as the type strains of these two groups, presented 99.8 and 99.1% similarity, respectively, in their 16S rRNA gene sequences with respect to B. elkanii USDA 76T. In spite of these high similarity values, the analysis of additional phylogenetic markers such as atpD and glnII genes and the 16S-23S intergenic spacer (ITS) showed that strains PAC48T and PAC68T represented two separate novel species of the genus Bradyrhizobium with B. elkanii as their closest relative. Phenotypic differences among the novel strains isolated from Pachyrhizus and B. elkanii were found regarding the assimilation of carbon sources and antibiotic resistance. All these differences were congruent with DNA-DNA hybridization analysis which revealed 21% genetic relatedness between strains PAC48T and PAC68T and 46% and 25%, respectively, between these strains and B. elkanii LMG 6134T. The nodD and nifH genes of strains PAC48T and PAC68T were phylogenetically divergent from those of bradyrhizobia species that nodulate soybean. Soybean was not nodulated by the novel Pachyrhizus isolates. Based on the genotypic and phenotypic data obtained in this study, the new strains represent two novel species for which the names Bradyrhizobium pachyrhizi sp. nov. (type strain PAC48T=LMG 24246T=CECT 7396T) and Bradyrhizobium jicamae sp. nov. (type strain PAC68T=LMG 24556T=CECT 7395T) are proposed.

  3. A novel chalcone derivative from Onychium japonicum

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A novel chalcone derivative was isolated from Onychium japonicum and its structure was elucidated by spectroscopic methods.The isolated compound showed cytotoxic activity against the human carcinoma cell lines Hela and BEL-7402 in vitro.(C) 2007 Hong Quan Duan. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

  4. 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.

  5. 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

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

    Directory of Open Access Journals (Sweden)

    Mieczysława Deryło

    2014-01-01

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

  7. 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

    chromatography and conventional mass spectrometry (MS) methods, including MS-mass spectroscopy, UV spectroscopy, and high-performance liquid chromatography-MS. The identified products indicate a novel metabolic pathway in which IAA is metabolized via dioxindole-3-acetic acid, dioxindole, isatin, and 2......-aminophenyl glyoxylic acid (isatinic acid) to anthranilic acid, which is further metabolized. Degradation of 4-Cl-IAA apparently stops at the 4-Cl-dioxindole step in contrast to 5-Cl-IAA which is metabolized to 5-Cl-anthranilic acid. Udgivelsesdato: 1995-Oct...

  8. 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.

  9. Genome Sequence of Bradyrhizobium viridifuturi Strain SEMIA 690T, a Nitrogen-Fixing Symbiont of Centrosema pubescens

    Science.gov (United States)

    Helene, Luisa Caroline Ferraz; Gomes, Douglas Fabiano; Delamuta, Jakeline Renata Marçon; Ribeiro, Renan Augusto; Souza, Renata Carolini; Almeida, Luiz Gonzaga Paula; Vasconcelos, Ana Tereza Ribeiro

    2015-01-01

    SEMIA 690T is a nitrogen-fixing symbiont of Centrosema pubescens, and comprises the recently described species Bradyrhizobium viridifuturi. Its draft genome indicates that it belongs to the Bradyrhizobium elkanii superclade. SEMIA 690T carries two copies of the regulatory nodD gene, and the nod and nif operons resemble those of Bradyrhizobium diazoefficiens. PMID:26679590

  10. Draft Genome Sequence of Bradyrhizobium elkanii TnphoA 33, a Producer of Polyhydroxyalkanoates

    Science.gov (United States)

    Lopes, Erica M.; Kishi, Luciano T.; Fernandes, Camila C.; Paganelli, Fernanda Larozza; Alves, Lucia M. C.; de Souza, Jackson A. Marcondes

    2017-01-01

    ABSTRACT The genus Bradyrhizobium comprises bacteria with the ability to form nitrogen-fixing symbioses with legumes. They are of great interest in agriculture, as well as for the production of biopolymers such as polyhydroxyalkanoates. Here, we report the draft genome assembly of Bradyrhizobium elkanii TnphoA 33 comprising 9 Mb, 1,124 contigs, and 9,418 open reading frames. PMID:28232432

  11. Genome Sequence of Bradyrhizobium pachyrhizi Strain PAC48T, a Nitrogen-Fixing Symbiont of Pachyrhizus erosus (L.) Urb.

    Science.gov (United States)

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

    2015-09-17

    Bradyrhizobium pachyrhizi PAC48(T) has been isolated from a jicama nodule in Costa Rica. The draft genome indicates high similarity with that of Bradyrhizobium elkanii. Several coding sequences (CDSs) of the stress response might help in survival in the tropics. PAC48(T) carries nodD1 and nodK, similar to Bradyrhizobium (Parasponia) ANU 289 and a particular nodD2 gene.

  12. 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

  13. 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

    2016-10-14

    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.

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

    Science.gov (United States)

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

    2009-06-01

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

  15. 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.

  16. 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.

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

    OpenAIRE

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

    2011-01-01

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

  18. 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%.

  19. 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

  20. 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.

  1. 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.

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

    Science.gov (United States)

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

  3. 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.

  4. Complete Genome Sequence of Bradyrhizobium diazoefficiens USDA 122, a Nitrogen-Fixing Soybean Symbiont

    Science.gov (United States)

    Sugawara, Masayuki; Tsukui, Takahiro; Kaneko, Takakazu; Ohtsubo, Yoshiyuki; Sato, Shusei; Nagata, Yuji; Tsuda, Masataka; Mitsui, Hisayuki

    2017-01-01

    ABSTRACT We report the complete genome sequence of Bradyrhizobium diazoefficiens USDA 122, a nitrogen-fixing soybean symbiont. The genome consists of a 9.1 Mb circular chromosome, and 8,551 coding sequences (CDSs) were predicted on the genome. The sequence will provide insight into the evolution of rhizobial genome, and the symbiotic compatibility with host plants. PMID:28254989

  5. Cloning and characterisation of Schistosoma japonicum insulin receptors.

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    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

  6. Bradyrhizobium manausense sp. nov., isolated from effective nodules of Vigna unguiculata grown in Brazilian Amazonian rainforest soils.

    Science.gov (United States)

    Silva, Flavia V; De Meyer, Sofie E; Simões-Araújo, Jean L; Barbé, Tatiane da Costa; Xavier, Gustavo R; O'Hara, Graham; Ardley, Julie K; Rumjanek, Norma G; Willems, Anne; Zilli, Jerri E

    2014-07-01

    Root nodule bacteria were trapped within cowpea (Vigna unguiculata) in soils with different cultivation histories collected from the Amazonian rainforest in northern Brazil. Analysis of the 16S rRNA gene sequences of six strains (BR 3351(T), BR 3307, BR 3310, BR 3315, BR 3323 BR and BR 3361) isolated from cowpea nodules showed that they formed a distinct group within the genus Bradyrhizobium, which was separate from previously identified type strains. Phylogenetic analyses of three housekeeping genes (glnII, recA and rpoB) revealed that Bradyrhizobium huanghuaihaiense CCBAU 23303(T) was the most closely related type strain (96% sequence similarity or lower). Chemotaxonomic data, including fatty acid profiles (predominant fatty acids being C16 : 0 and summed feature 8), the slow growth rate and carbon compound utilization patterns supported the assignment of the strains to the genus Bradyrhizobium. The results of DNA-DNA hybridizations, antibiotic resistance and physiological tests differentiated these novel strains from the most closely related species of the genus Bradyrhizobium with validly published names. Symbiosis-related genes for nodulation (nodC) and nitrogen fixation (nifH) grouped the novel strains of the genus Bradyrhizobium together with Bradyrhizobium iriomotense strain EK05(T), with 94% and 96% sequence similarity, respectively. Based on these data, these six strains represent a novel species for which the name Brabyrhizobium manausense sp. nov. (BR 3351(T) = HAMBI 3596(T)), is proposed.

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

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    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.

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

    DEFF Research Database (Denmark)

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

    2016-01-01

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

  9. Genome sequence of the Ornithopus/Lupinus-nodulating Bradyrhizobium sp. strain WSM471.

    Science.gov (United States)

    Reeve, Wayne; De Meyer, Sofie; Terpolilli, Jason; Melino, Vanessa; Ardley, Julie; Tian, Rui; Tiwari, Ravi; Howieson, John; Yates, Ronald; O'Hara, Graham; Ninawi, Mohamed; Lu, Megan; Bruce, David; Detter, Chris; Tapia, Roxanne; Han, Cliff; Wei, Chia-Lin; Huntemann, Marcel; Han, James; Chen, I-Min; Mavromatis, Konstantinos; Markowitz, Victor; Ivanova, Natalia; Pagani, Ioanna; Pati, Amrita; Goodwin, Lynne; Woyke, Tanja; Kyrpides, Nikos

    2013-12-20

    Bradyrhizobium sp. strain WSM471 is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from an effective nitrogen- (N2) fixing root nodule formed on the annual legume Ornithopus pinnatus (Miller) Druce growing at Oyster Harbour, Albany district, Western Australia in 1982. This strain is in commercial production as an inoculant for Lupinus and Ornithopus. Here we describe the features of Bradyrhizobium sp. strain WSM471, together with genome sequence information and annotation. The 7,784,016 bp high-quality-draft genome is arranged in 1 scaffold of 2 contigs, contains 7,372 protein-coding genes and 58 RNA-only encoding genes, and is one of 20 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Community Sequencing Program.

  10. 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.

  11. 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.

  12. 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

  13. Genome sequence of the lupin-nodulating Bradyrhizobium sp. strain WSM1417.

    Science.gov (United States)

    Reeve, Wayne; Terpolilli, Jason; Melino, Vanessa; Ardley, Julie; Tian, Rui; De Meyer, Sofie; Tiwari, Ravi; Yates, Ronald; O'Hara, Graham; Howieson, John; Ninawi, Mohamed; Teshima, Hazuki; Bruce, David; Detter, Chris; Tapia, Roxanne; Han, Cliff; Wei, Chia-Lin; Huntemann, Marcel; Han, James; Chen, I-Min; Mavrommatis, Konstantinos; Markowitz, Victor; Ivanova, Natalia; Ovchinnikova, Galina; Pagani, Ioanna; Pati, Amrita; Goodwin, Lynne; Peters, Lin; Woyke, Tanja; Kyrpides, Nikos

    2013-12-20

    Bradyrhizobium sp. strain WSM1417 is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated from an effective nitrogen (N2) fixing root nodule of Lupinus sp. collected in Papudo, Chile, in 1995. However, this microsymbiont is a poorly effective N2 fixer with the legume host Lupinus angustifolius L.; a lupin species of considerable economic importance in both Chile and Australia. The symbiosis formed with L. angustifolius produces less than half of the dry matter achieved by the symbioses with commercial inoculant strains such as Bradyrhizobium sp. strain WSM471. Therefore, WSM1417 is an important candidate strain with which to investigate the genetics of effective N2 fixation in the lupin-bradyrhizobia symbioses. Here we describe the features of Bradyrhizobium sp. strain WSM1417, together with genome sequence information and annotation. The 8,048,963 bp high-quality-draft genome is arranged in a single scaffold of 2 contigs, contains 7,695 protein-coding genes and 77 RNA-only encoding genes, and is one of 20 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Community Sequencing Program.

  14. Triple inoculation with Bradyrhizobium, Glomus and Paenibacillus on cowpea (Vigna unguiculata [L.] walp.) development.

    Science.gov (United States)

    de Lima, André Suêldo Tavares; Xavier, Terezinha Ferreirab; de Lima, Cláudia Elizabete Pereira; de Paula Oliveira, José; Mergulhão, Adália Cavalcanti do Espírito Santo; Figueiredo, Figueiredo Márcia do Vale Barreto

    2011-07-01

    The use of microorganisms to improve the availability of nutrients to plants is of great importance to agriculture. This study aimed to evaluate the effect of triple inoculation of cowpea with arbuscular mycorrhizal fungi (AMF), plant growth-promoting bacteria (PGPB) and rhizobia to maximize biological nitrogen fixation (BNF) and promote plant growth. The experiment was conducted in a greenhouse using cowpea plants (Vigna unguiculata L. Walp cv. IPA 206). The treatments included inoculation with strains of Bradyrhizobium sp. (BR 3267 and EI - 6) individually and as a mixture, an absolute control (AC) and mineral nitrogen control (NC), all combined with the presence or absence of native AMF (Glomus etunicatum) and PGPB (Paenibacillus brasilensis - 24) in a 5x2x2 factorial design. All treatments were replicated three times. Contrasts were performed to study the treatment of variables. Inoculation with Bradyrhizobium sp. (BR 3267 and EI - 6) and G. etunicatum favored nitrogen acquisition and phosphorus availability for the cowpea plants. Inoculation with P. brasilensis - 24 increased colonization by Bradyrhizobium sp. and G. etunicatum and promoted cowpea growth, while the nitrogen from symbiosis was sufficient to supply the plants nutritional needs.

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

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    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.

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

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    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

  17. 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.

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

    Science.gov (United States)

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

    2015-07-27

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

  19. 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.

  20. 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.

  1. First Report of Leaf Rust Caused by Puccinia caricis in Farfugium japonicum in Korea.

    Science.gov (United States)

    Yun, Yeo Hong; Kwon, Hyuk Woo; Ahn, Hong Seok; Kim, Seong Hwan

    2015-09-01

    Farfugium japonicum is used in traditional medicine and as an edible herb in China and Korea. In July 2013, leaf spots were observed in F. japonicum seedlings at Ulleung Island, Gyeongsangbuk Province, Korea. Early symptoms on the leaf adaxial surface included roughly circular yellow spots that later developed brown, necrotic centers. The aecia were hypophyllous, cupulate, yellowish, 180~430 µm in diameter, clustered, and erumpent with a peridium with a recurved margin. The aeciospores were globoid, 14~17 × 13~16 µm, light yellow or colorless, and densely verrucose. The 28S rDNA sequence of the isolate was identical to each other and shared 99% identity with Puccinia caricis. This is the first report of rust caused by P. caricis in F. japonicum in Korea or elsewhere in the world.

  2. 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.

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

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    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. 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.

  5. 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.

  6. Lethal and sublethal effects of thiamethoxam on the whitefly predator Serangium japonicum (Coleoptera: Coccinellidae) through different exposure routes.

    Science.gov (United States)

    Yao, Feng-Luan; Zheng, Yu; Zhao, Jian-Wei; Desneux, Nicolas; He, Yu-Xian; Weng, Qi-Yong

    2015-06-01

    Given expectations for a booming usage of thiamethoxam and increasing availability of the promising biological agent Serangium japonicum for the control of Bemisia tabaci in China, an evaluation of their compatibility is crucial for integrated pest management (IPM). This study examined the lethal and sublethal effects of thiamethoxam on S. japonicum through three exposure routes. An acute toxicity bioassay showed that LC50 values of thiamethoxam for S. japonicum through residue contact, egg-dip, and systemic treatment were 6.65, 4.37, and 2.43 mg AI L(-1), respectively. The prey consumption of S. japonicum given different densities of B. tabaci eggs under control, discontinuous, egg-dip and systemic exposure scenarios showed a good fit to a Type II functional response. Predation of S. japonicum was most affected under systemic exposure, followed by egg-dip, and discontinuous, which was only slightly affected. In all cases tested, however, predators recovered their predation capacity rapidly, either after 24h of exposure or 24h after the end of exposure. Thiamethoxam was highly toxic to S. japonicum regardless of exposure routes. Sublethal effects of thiamethoxam applied systemically or foliar both impaired the biological control of S. japonicum on B. tabaci. Therefore, thiamethoxam should be used with caution in IPM of B. tabaci.

  7. 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.

  8. 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

  9. Developmental gene expression profiles of the human pathogen Schistosoma japonicum

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    McManus Donald P

    2009-03-01

    Full Text Available Abstract Background The schistosome blood flukes are complex trematodes and cause a chronic parasitic disease of significant public health importance worldwide, schistosomiasis. Their life cycle is characterised by distinct parasitic and free-living phases involving mammalian and snail hosts and freshwater. Microarray analysis was used to profile developmental gene expression in the Asian species, Schistosoma japonicum. Total RNAs were isolated from the three distinct environmental phases of the lifecycle – aquatic/snail (eggs, miracidia, sporocysts, cercariae, juvenile (lung schistosomula and paired but pre-egg laying adults and adult (paired, mature males and egg-producing females, both examined separately. Advanced analyses including ANOVA, principal component analysis, and hierarchal clustering provided a global synopsis of gene expression relationships among the different developmental stages of the schistosome parasite. Results Gene expression profiles were linked to the major environmental settings through which the developmental stages of the fluke have to adapt during the course of its life cycle. Gene ontologies of the differentially expressed genes revealed a wide range of functions and processes. In addition, stage-specific, differentially expressed genes were identified that were involved in numerous biological pathways and functions including calcium signalling, sphingolipid metabolism and parasite defence. Conclusion The findings provide a comprehensive database of gene expression in an important human pathogen, including transcriptional changes in genes involved in evasion of the host immune response, nutrient acquisition, energy production, calcium signalling, sphingolipid metabolism, egg production and tegumental function during development. This resource should help facilitate the identification and prioritization of new anti-schistosome drug and vaccine targets for the control of schistosomiasis.

  10. 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.

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

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    Xiaojun Chen

    2014-05-01

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

  12. 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.

  13. Expression of symbiotic genes of Rhizobium japonicum USDA 191 in other rhizobia.

    OpenAIRE

    Appelbaum, E R; McLoughlin, T J; O'Connell, M; Chartrain, N

    1985-01-01

    A 200-megadalton plasmid was mobilized from Rhizobium japonicum USDA 191 to other Rhizobium strains either that cannot nodulate soybeans or that form Fix- nodules on certain cultivars. The symbiotic properties of the transconjugants indicate that both soybean specificity for nodulation and cultivar specificity for nitrogen fixation are plasmid encoded.

  14. Co-dispersal of the blood fluke Schistosoma japonicum and Homo sapiens in the Neolithic Age.

    Science.gov (United States)

    Yin, Mingbo; Zheng, Hong-Xiang; Su, Jing; Feng, Zheng; McManus, Donald P; Zhou, Xiao-Nong; Jin, Li; Hu, Wei

    2015-12-21

    The global spread of human infectious diseases is of considerable public health and biomedical interest. Little is known about the relationship between the distribution of ancient parasites and that of their human hosts. Schistosoma japonicum is one of the three major species of schistosome blood flukes causing the disease of schistosomiasis in humans. The parasite is prevalent in East and Southeast Asia, including the People's Republic of China, the Philippines and Indonesia. We studied the co-expansion of S. japonicum and its human definitive host. Phylogenetic reconstruction based on complete mitochondrial genome sequences showed that S. japonicum radiated from the middle and lower reaches of the Yangtze River to the mountainous areas of China, Japan and Southeast Asia. In addition, the parasite experienced two population expansions during the Neolithic agriculture era, coinciding with human migration and population growth. The data indicate that the advent of rice planting likely played a key role in the spread of schistosomiasis in Asia. Moreover, the presence of different subspecies of Oncomelania hupensis intermediate host snails in different localities in Asia allowed S. japonicum to survive in new rice-planting areas, and concurrently drove the intraspecies divergence of the parasite.

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

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    Jian Huang

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

  16. 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.

  17. 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.

  18. 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.

  19. 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.

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

    Science.gov (United States)

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

    2017-02-08

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

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

    Directory of Open Access Journals (Sweden)

    Rui Tang

    2017-02-01

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

  2. Draft genome sequence of Bradyrhizobium sp. strain BR 3262, an effective microsymbiont recommended for cowpea inoculation in Brazil

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    Jean Luiz Simões-Araújo

    Full Text Available Abstract The strain BR 3262 was isolated from nodule of cowpea (Vigna unguiculata L. Walp growing in soil of the Atlantic Forest area in Brazil and it is reported as an efficient nitrogen fixing bacterium associated to cowpea. Firstly, this strain was assigned as Bradyrhizobium elkanii, however, recently a more detailed genetic and molecular characterization has indicated it could be a Bradyrhizobium pachyrhizi species. We report here the draft genome sequence of B. pachyrhizi strain BR 3262, an elite bacterium used as inoculant for cowpea. The whole genome with 116 scaffolds, 8,965,178 bp and 63.8% of C+G content for BR 3262 was obtained using Illumina MiSeq sequencing technology. Annotation was added by the RAST prokaryotic genome annotation service and shown 8369 coding sequences, 52 RNAs genes, classified in 504 subsystems.

  3. Draft genome sequence of Bradyrhizobium sp. strain BR 3262, an effective microsymbiont recommended for cowpea inoculation in Brazil.

    Science.gov (United States)

    Simões-Araújo, Jean Luiz; Leite, Jakson; Marie Rouws, Luc Felicianus; Passos, Samuel Ribeiro; Xavier, Gustavo Ribeiro; Rumjanek, Norma Gouvêa; Zilli, Jerri Édson

    The strain BR 3262 was isolated from nodule of cowpea (Vigna unguiculata L. Walp) growing in soil of the Atlantic Forest area in Brazil and it is reported as an efficient nitrogen fixing bacterium associated to cowpea. Firstly, this strain was assigned as Bradyrhizobium elkanii, however, recently a more detailed genetic and molecular characterization has indicated it could be a Bradyrhizobium pachyrhizi species. We report here the draft genome sequence of B. pachyrhizi strain BR 3262, an elite bacterium used as inoculant for cowpea. The whole genome with 116 scaffolds, 8,965,178bp and 63.8% of C+G content for BR 3262 was obtained using Illumina MiSeq sequencing technology. Annotation was added by the RAST prokaryotic genome annotation service and shown 8369 coding sequences, 52 RNAs genes, classified in 504 subsystems.

  4. 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

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

    OpenAIRE

    Lardi, Martina; Murset, Valérie; Fischer, Hans-Martin; Mesa, Socorro; Ahrens, Christian H.; Zamboni, Nicola; Pessi, Gabriella

    2016-01-01

    Bradyrhizobium diazoefficiens is a nitrogen-fixing endosymbiont, which can grow inside root-nodule cells of the agriculturally important soybean and other host plants. Our previous studies described B. diazoefficiens host-specific global expression changes occurring during legume infection at the transcript and protein level. In order to further characterize nodule metabolism, we here determine by flow injection–time-of-flight mass spectrometry analysis the metabolome of (i) nodules and roots...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1990-07-01

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

  7. Does multiple hosts mean multiple parasites? Population genetic structure of Schistosoma japonicum between definitive host species.

    Science.gov (United States)

    Wang, T P; Shrivastava, J; Johansen, M V; Zhang, S Q; Wang, F F; Webster, J P

    2006-10-01

    Multi-host parasites, those capable of infecting more than one species of host, are responsible for the majority of all zoonotic, emerging or persistent human and animal diseases and are considered one of the major challenges for the biomedical sciences in the 21st century. We characterized the population structure of the multi-host parasite Schistosoma japonicum in relation to its definitive host species by genotyping miracidia collected from humans and domestic animals across five villages around the Yangtze River in Anhui Province, mainland China, using microsatellite markers. High levels of polymorphisms were observed and two main genetic clusters were identified which separated water buffalo, cattle and humans from goats, pigs, dogs and cats. We thereby believe that we present the first evidence of definitive host-based genetic variation in Schistosoma japonicum which has important epidemiological, evolutionary, medical and veterinary implications.

  8. 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.

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

    Science.gov (United States)

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

    2014-12-12

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

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

    DEFF Research Database (Denmark)

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

    2007-01-01

    The pig is a natural host for Schistosoma japonicum and a useful animal model of human infection. The aim of the present study was to assess the differences between the cytokine profiles in prenatally or postnatally S. japonicum exposed pigs. Seven prenatally exposed pigs, 7 postnatally exposed...... pigs and 4 uninfected control pigs were compared 27 weeks post exposure. Variables included worm burdens, tissue egg counts, liver pathology and mRNA levels of IL-2, IL-4, IL-10, IL-12, TNF-alpha, TGF-beta 1 and IFN-gamma in the liver and the caecum, assessed by RT-PCR. Infection intensity and level...... of septal fibrosis were significantly higher in the postnatal group compared to the prenatal group (P TNF-alpha (P

  11. 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

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

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    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.

  13. A circular analysis of chronobiology of Schistosoma japonicum cercarial emergence from hilly areas of Anhui, China.

    Science.gov (United States)

    Su, Jing; Zhou, Feng; Lu, Da-Bing

    2013-10-01

    About 46 mammal species have been suspected as reservoir hosts for Schistosoma japonicum and therefore the track of the target parasites, in relation to definitive host species, may be of great importance in terms of theoretical and practical implications. The circadian rhythm of cercariae emergence, a genetically controlled behavior for parasites to adapt to their definitive hosts, may seem to be a perfect biological marker for S. japonicum. In this study, a late (or nocturnal) cercarial emergence pattern was observed on the parasites from one hilly region in Anhui of China, where rodents serve as reservoirs, and on the first generation of the parasites. Moreover, by using the circular statistics, the homogeneity of parasites in such trait was also demonstrated. All these provide evidence for the genetically controlled biological trait, which seems essential in the investigation of macro- or micro-dynamics of parasite transmission of interest. This is particularly true in the case of S. japonicum when multiple parasite isolates or strains are more likely to exist.

  14. 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.

  15. 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.

  16. 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.

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

    Directory of Open Access Journals (Sweden)

    2006-04-01

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

  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. 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.

  20. Sinergismo Bacillus, Brevibacillus e, ou, Paenibacillus na simbiose Bradyrhizobium-caupi

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    André Suêldo Tavares de Lima

    2011-06-01

    Full Text Available O feijão-caupi Vigna unguiculata [L.] Walp. é a principal cultura de subsistência do semiárido brasileiro, sendo fonte de proteínas de baixo custo, notadamente, para populações carentes. A produção dessa cultura no Nordeste é baixa devido à não utilização de insumos agrícolas - entre eles, o fertilizante nitrogenado. Por outro lado, bactérias promotoras de crescimento em plantas (BPCPs vêm sendo estudadas, de forma a maximizar a fixação de N2, disponibilizar nutrientes como P ou fito-hormônios e inibir doenças. Os objetivos deste trabalho foram verificar a viabilidade da coinoculação das sementes de feijão-caupi usando Paenibacillus, Brevibacillus e, ou, Bacillus e Bradyrhizobium-caupi; caracterizar as estirpes quanto à produção de ácido indol acético (AIA e solubilização de fosfato; e avaliar o sinergismo entre os microrganismos como alternativa para otimizar a FBN. Os experimentos foram conduzidos em laboratório e em casa de vegetação do Instituto Agronômico de Pernambuco (IPA, utilizando o feijão-caupi cv. "IPA - 206". As estirpes utilizadas foram: Bacillus sp. - 449, 450, 451, 461 e ANBE 31; B. cereus - 440; B. subtilis - 438, 441, 454, 455 e 459; B. pumilus - 444, 445 e 448; B. megaterium - 462; Brevibacillus brevis - 447; Paenibacillus brasilensis - 24, 172 e 177; P. graminis - MC 04.21, MC 22.13 e BR 60106; P. polymyxa - S21; e P. durus - RBN4. Os microrganismos não apresentaram capacidade para produzir AIA nem solubilizar fosfato. Ocorreu sinergismo das estirpes de Bacillus sp. (449 e Bacillus pumilus (444 coinoculadas com a estirpe de Bradyrhizobium sp. (BR 3267 no feijão-caupi.

  1. Competitiveness of a Bradyrhizobium sp. strain in soils containing indigenous rhizobia.

    Science.gov (United States)

    Bogino, Pablo; Banchio, Erika; Bonfiglio, Carlos; Giordano, Walter

    2008-01-01

    The success of rhizobial inoculation on plant roots is often limited by several factors, including environmental conditions, the number of infective cells applied, the presence of competing indigenous (native) rhizobia, and the inoculation method. Many approaches have been taken to solve the problem of inoculant competition by naturalized populations of compatible rhizobia present in soil, but so far without a satisfactory solution. We used antibiotic resistance and molecular profiles as tools to find a reliable and accurate method for competitiveness assay between introduced Bradyrhizobium sp. strains and indigenous rhizobia strains that nodulate peanut in Argentina. The positional advantage of rhizobia soil population for nodulation was assessed using a laboratory model in which a rhizobial population is established in sterile vermiculite. We observed an increase in nodule number per plant and nodule occupancy for strains established in vermiculite. In field experiments, only 9% of total nodules were formed by bacteria inoculated by direct coating of seed, whereas 78% of nodules were formed by bacteria inoculated in the furrow at seeding. In each case, the other nodules were formed by indigenous strains or by both strains (inoculated and indigenous). These findings indicate a positional advantage of native rhizobia or in-furrow inoculated rhizobia for nodulation in peanut.

  2. 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.

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

    Science.gov (United States)

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

    2014-12-01

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

  4. 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

  5. Estirpes de Paenibacillus promotoras de nodulação específica na simbiose Bradyrhizobium-caupi = Strains of Paenibacillus promoters of the specific nodulation in the symbiosis Bradyrhizobium-caupi

    Directory of Open Access Journals (Sweden)

    Valéria Nogueira da Silva

    2007-07-01

    Full Text Available O efeito de estirpes de Paenibacillus e Bacillus (Rizobactérias Promotoras de Crescimento de Plantas - RPCP na simbiose do caupi [Vigna unguiculata (L. Walp.] com Bradyrhizobium sp. foi estudado em vaso de Leonard. A cultivar de caupi utilizada foi a IPA-205. As plantas foram irrigadas com solução nutritiva sem nitrogênio. As inoculações com RPCP promoveram maior crescimento da raiz em relação às co-inoculações, como a estirpe S21 de P. polymyxa, que foi superior (70% ao controle (não-inoculado. As RPCP aumentaram a nodulação específica nas plantas co-inoculadas, evidenciado pela correlação positiva com o nitrogênio acumulado específico. Efeitos indiretos e diretos na nodulação específica foram mostrados pela diminuição significativa do crescimento da raiz (40% e pelo aumento não-significativo do número de nódulos (46%, nas estirpes Loutit (L de P.polymyxa e na LBF-410 de P. macerans. RPCP que estimulam a nodulação podem promover uma melhor fixação de N2 durante a simbiose do Bradyrhizobium-caupi.The effect of Paenibacillus and Bacillus (Plant Growth Promoting Rhizobacteria - PGPR strains on the symbiosis of the caupi [Vigna unguiculata (L. Walp.] with Bradyrhizobium sp. was studied in vases of Leonard. The caupi cultivar used was IPA-205. The plants were irrigated with an N-Free nutrient solution. The inoculations with PGPR promoted a better growth of the root in relation to the o-inoculations, as the S21 strain of P. polymyxa which was superior (70% to the control (not inoculated. The PGPR increased the specific nodulation in the co-inoculated plants, evidenced by thepositive correlation with the specific nitrogen accumulated. Indirect and direct effects on the specific nodulation were shown by the significant decrease of root growth (40% and non-significant increase in the number of nodules (46% in the Loutit (L strain of P.polymyxa and LBF-410 strain of P. macerans. The PGPR, which estimulated the nodulation

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

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    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. Molecular cloning and characterization of glutamine synthetase, a tegumental protein from Schistosoma japonicum.

    Science.gov (United States)

    Qiu, Chunhui; Hong, Yang; Cao, Yan; Wang, Fei; Fu, Zhiqiang; Shi, Yaojun; Wei, Meimei; Liu, Shengfa; Lin, Jiaojiao

    2012-12-01

    Glutamine synthetase catalyzes the synthesis of glutamine, providing nitrogen for the production of purines, pyrimidines, amino acids, and other compounds required in many pivotal cellular events. Herein, a full-length cDNA encoding Schistosoma japonicum glutamine synthetase (SjGS) was isolated from 21-day schistosomes. The entire open reading frame of SjGS contains a 1,095-bp coding region corresponding to 364 amino acids with a calculated molecular weight of 40.7 kDa. NCBIP blast shows that the putative amino acid of SjGS contains a classic β-grasp domain and a catalytic domain of glutamine synthetase. The relative mRNA expression of SjGS was evaluated in 7-, 13-, 21-, 28-, 35-, and 42-day worms of S. japonicum in the final host and higher expression at day 21, and 42 worms were observed. This protein was also detected in worm extracts using Western blot. Immunofluorescence studies indicated that the SjGS protein was mainly distributed on tegument and parenchyma in 28-day adult worms. The recombinant glutamine synthetase with a molecular weight of 45 kDa was expressed in Escherichia coli and purified in its active form. The enzyme activity of the recombinant protein was 3.30 ± 0.67 U.μg-1. The enzyme activity was highly stable over a wide range of pH (6-9) and temperature (25-40 °C) under physiological conditions. The transcription of SjGS was upregulated in praziquantel-treated worms at 2-, 4-, and 24-h posttreatment compared with the untreated control. As a first step towards the clarification of the role of glutamine synthetase in schistosome species, we have cloned and characterized cDNAs encoding SjGS in S. japonicum, and the data presented suggest that SjGS is an important molecule in the development of the schistosome.

  8. A strategy for emergency treatment of Schistosoma japonicum-infested water

    Directory of Open Access Journals (Sweden)

    Li Hong-Jun

    2011-11-01

    Full Text Available Abstract Background Schistosomiasis japonica, caused by contact with Schistosoma japonicum cercaria-infested water when washing, bathing or production, remains a major public-health concern in China. The purpose of the present study was to investigate the effect of a suspension concentrate of niclosamide (SCN on killing cercaria of S. japonicum that float on the water surface, and its toxicity to fish, so as to establish an emergency-treatment intervention for rapidly killing cercaria and eliminating water infectivity. Results At 30 min after spraying 100 mg/L SCN, with niclosamide dosages of 0.01, 0.02, 0.03, 0.04 g/m2, the water infectivity reduced significantly and no infectivity was found at 60 min after spraying SCN. The surface of static water was sprayed with 100 mg/L SCN, the peak concentration was found at 0 min, and the solution diffused to site with a water depth of 10 cm after 10 min. 30 min later, SCN diffused to the whole water body, and distributed evenly. After spraying 100 mg/L SCN onto the surface of the water with a volume of(3.14 × 202×50cm3, with niclosamide dosages of 0.02 g/m2, 96 h later, no death of zebra fish was observed. Conclusions By spraying 100 mg/L SCN, with a niclosamide dosage of 0.02 g/m2 onto the surface of S. japonicum-infested water, infectivity of the water can be eliminated after 30-60 min, and there is no evident toxicity to fish. This cercaria-killing method, as an emergency-treatment intervention for infested water, can be applied in those forecasting and early warning systems for schistosomiasis.

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

    OpenAIRE

    J. A. Freixas; Inés M. Reynaldo; María C. Nápoles

    2010-01-01

    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 g...

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

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    Rachel J Stephenson

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

  11. An IL-13 promoter polymorphism associated with liver fibrosis in patients with Schistosoma japonicum.

    Science.gov (United States)

    Long, Xin; Chen, Qian; Zhao, Jianping; Rafaels, Nicholas; Mathias, Priyanka; Liang, Huifang; Potee, Joseph; Campbell, Monica; Zhang, Bixiang; Gao, Li; Georas, Steve N; Vercelli, Donata; Beaty, Terri H; Ruczinski, Ingo; Mathias, Rasika; Barnes, Kathleen C; Chen, Xiaoping

    2015-01-01

    The aim of this study was to determine whether two polymorphisms in the gene encoding IL13 previously associated with Schistosoma hematobium (S. hematobium) and S. mansoni infection are associated with S. japonicum infection. Single nucleotide polymorphisms (SNPs) rs1800925 (IL13/-1112C>T) and rs20541 (IL13R130Q) were genotyped in 947 unrelated individuals (307 chronically infected, 339 late-stage with liver fibrosis, 301 uninfected controls) from a schistosomiasis-endemic area of Hubei province in China. Regression models were used to evaluate allelic and haplotypic associations with chronic and late-stage schistosomiasis adjusted for non-genetic covariates. Expression of IL-13 was measured in S. japonicun-infected liver fibrosis tissue and normal liver tissue from uninfected controls by immunohistochemistry (IHC). The role of rs1800925 in IL-13 transcription was further determined by Luciferase report assay using the recombinant PGL4.17-rs180092 plasmid. We found SNP rs1800925T was associated with late-stage schistosomiasis caused by S. japonicum but not chronic schistosomiasis (OR = 1.39, 95%CI = 1.02-1.91, p = 0.03) and uninfected controls (OR = 1.49, 95%CI = 1.03-2.13, p = 0.03). Moreover, the haplotype rs1800925T-rs20541C increased the risk of disease progression to late-stage schistosomiasis (OR = 1.46, p = 0.035), whereas haplotype rs1800925C-rs20541A showed a protective role against development of late-stage schistosomiasis (F = 0.188, OR = 0.61, p = 0.002). Furthermore, S. japonicum-induced fibrotic liver tissue had higher IL13 expression than normal liver tissue. Plasmid PGL4.17-rs1800925T showed a stronger relative luciferase activity than Plasmid PGL4.17-rs1800925C in 293FT, QSG-7701 and HL-7702 cell lines. In conclusion, the functional IL13 polymorphism, rs1800925T, previously associated with risk of schistosomiasis, also contributes to risk of late-stage schistosomiasis caused by S. japonicum.

  12. Biofertilización con Bradyrhizobium japonicum para la agricultura sustentable: aspectos ecofisiológicos del problema de la competición para la nodulación

    OpenAIRE

    Covelli, Julieta Mariana

    2013-01-01

    El cultivo de soja es de gran importancia en nuestro país, sin embargo requiere un aporte importante de N para obtener buenos rendimientos, el cual, si se quiere promover la agricultura sustentable a largo plazo, no debería salir exclusivamente de la fertilidad del suelo, ni aún siendo suplementado por fertilizantes químicos. Una manera de abastecer N de forma sustentable y económica para lograr altos rindes podría ser la utilización del potencial de los rizobios fijadores de N2. Así, la biof...

  13. Eficácia da inoculação de Bradyrhizobium em pré-semeadura da soja Effectiveness of Bradyrhizobium inoculation at pre-sowing of soybean

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    Jerri Édson Zilli

    2010-03-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito da inoculação de Bradyrhizobium em pré-semeadura de soja e o efeito do tratamento das sementes com fungicidas na nodulação. Foram usados controles sem inoculação, que receberam ou não a aplicação de 200 kg ha-1 de N. A inoculação em pré-semeadura mostrou desempenho igual à inoculação padrão quanto à nodulação, produção de matéria seca, produtividade de grãos e acúmulo de nitrogênio nos tecidos e grãos da soja. Entretanto, quando as sementes foram tratadas com fungicidas, houve redução da nodulação e da produtividade.The objective of this work was to evaluate the effect of Bradyrhizobium inoculation on soybean pre-sowing and the effect of the treatments of seeds using fungicides on nodulation. Control treatments consisted of two non-inoculated treatments, with or without N-fertilizer supply (200 kg ha-1 of N. Inoculation at pre-sowing had a performance similar to the standard inoculation at sowing regarding soybean nodulation, dry matter production, grain yield and N accumulation in tissues and grains. However, when seeds were treated with fungicides, nodulation and grain yield decreased.

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

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    Giovanna Raso

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

  15. 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%).

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

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    Wang Heng

    2010-01-01

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

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

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    F.Sh.F. Badawi

    2011-06-01

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

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

    Science.gov (United States)

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

    2015-03-01

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

  19. 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.

  20. PRELIMINARY STUDY ON SCREENING OF EGG MIMOTOPE OF SCHISTOSOMA JAPONICUM%日本血吸虫模拟虫卵抗原的初步研究

    Institute of Scientific and Technical Information of China (English)

    钱旻; 白艳军; 章平; 吴自荣; 严自助

    2000-01-01

    @@ The egg of Schistosoma japonicum is a chief pathogenic factor. The preparation of egg antigen is a troublesome work, and needs a large number of experimental animals. In order to explore and develop an egg mimotope that can be used in the study of diagnostic reagents and vaccines, we employed the phage display random 15-peptides library to mimic the antigen epitope recognized by mAb 6B12 which is specific to egg antigen of Schistosoma japonicum.

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

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    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

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

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    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

  3. Evaluation of Immunoassays for the Diagnosis of Schistosoma japonicum Infection Using Archived Sera

    Science.gov (United States)

    Xu, Jing; Peeling, Rosanna W.; Chen, Jia-Xu; Wu, Xiao-Hua; Wu, Zhong-Dao; Wang, Shi-Ping; Feng, Ting; Chen, Shao-Hong; Li, Hao; Guo, Jia-Gang; Zhou, Xiao-Nong

    2011-01-01

    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, specificity

  4. Mefloquine in combination with hemin causes severe damage to adult Schistosoma japonicum in vitro.

    Science.gov (United States)

    Xiao, Shu-hua; Qiao, Chunhua; Xue, Jian; Wang, Lili

    2014-03-01

    In order to explore the interaction of mefloquine with hemin against adult Schistosoma japonicum in vitro, the 50% and 95% lethal concentration (LC50 and LC95) of mefloquine and hemin against schistosomes, some factors, such as other iron providing agents, iron chelaters, zinc protoporphyrin-IX, and biological relevant reductants, that might impact on antischistosomal activity induced by interaction of mefloquine with hemin, and preliminary analysis of chemical interaction of both compounds were undertaken. The LC50 and LC95 of mefloquine and hemin alone against schistosomes were determined to be 6.5μg/ml and 7.8μg/ml as well as 232μg/ml and 355μg/ml, respectively. The LC50 and LC95 of mefloquine in the presence of hemin 100μg/ml was 0.24μg/ml and 0.59μg/ml, respectively. On the other hand the LC50 and LC95 of hemin in the presence of mefloquine 1μg/ml was 23.2μg/ml and 77.2μg/ml, respectively. Meanwhile, mefloquine/hemin combinations showed potential synergistic effects against adult S. japonicum, with combination index (CI) values mefloquine exhibited no toxic effect against schistosomes. On the other hand, addition of iron chelators (deferiprone, desferrioxamine mesylate, or 2,2'-bipyridine) to the medium containing mefloquine-hemin resulted in no protective effect on the worms. Furthermore, biological reductants like glutathione, vitamine C or cysteine showed no apparent worm protection effect from toxic mefloquine-hemin even at higher concentrations (242.3-614.6μg/ml, i.e., 6.4-17.8-fold higher than the concentration of hemin). Chemical interaction of mefloquine with hemin was studied in 40% DMSO-Tris buffer solution. Both UV-Vis spectrum and mass spectrum demonstrated the strong interaction of mefloquine with hemin, which resulted in a reduction of hemin color and emergence of an adduct formed by mefloquine and hemin. The results confirm that mefloquine combined with hemin exhibits potential synergistic effect against adult S. japonicum in vitro.

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

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    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

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

    Directory of Open Access Journals (Sweden)

    Márcia Rufini

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1993-06-01

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

  8. 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.

  9. 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

    2015-10-12

    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.

  10. 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.

  11. Silver-enhanced colloidal gold metalloimmunoassay for Schistosoma japonicum antibody detection.

    Science.gov (United States)

    Chu, Xia; Xiang, Zhi-Feng; Fu, Xin; Wang, Shi-Ping; Shen, Guo-Li; Yu, Ru-Qin

    2005-06-01

    A silver-enhanced colloidal gold metalloimmunoassay has been proposed for the determination of Schistosoma japonicum antibody (SjAb) in rabbit serum. The adult worm antigen of S. japonicum (SjAg) was adsorbed passively on the walls of a polystyrene microwell and then reacted with the desired SjAb. The colloidal gold-labeled goat anti-rabbit IgG secondary antibody was adsorbed on the walls of the polystyrene microwells through the reaction with SjAb, followed by the silver enhancement process, dissolution of silver metal atoms in an acidic solution, and determination of dissolved silver ions by anodic stripping voltammetry (ASV) at a glassy-carbon electrode. Assay conditions were optimized, including the reaction time of SjAg with SjAb, the interaction of SjAb with the colloidal gold-labeled secondary antibody, the dilution ratio of the colloidal gold-labeled secondary antibody and the silver enhancement time. The integration of the anodic stripping peak current depended linearly on the SjAb logarithmic concentration over the range of 6.4 ng/ml to 100 microg/ml. A detection limit as low as 3.0 ng/ml SjAb was achieved, which was better than the piezoelectric body acoustic wave sensor (detection limit of 7.2 microg/ml) and the renewable amperometric immunosensor (detection limit of 0.36 microg/ml). Rabbit serum samples with various degrees of infection were analyzed, and the results demonstrate that the proposed method meets the requirements of clinical analysis.

  12. Genome sequence of Bradyrhizobium sp. WSM1253; a microsymbiont of Ornithopus compressus from the Greek Island of Sifnos.

    Science.gov (United States)

    Tiwari, Ravi; Howieson, John; Yates, Ron; Tian, Rui; Held, Britanny; Tapia, Roxanne; Han, Cliff; Seshadri, Rekha; Reddy, T B K; Huntemann, Marcel; Pati, Amrita; Woyke, Tanja; Markowitz, Victor; Ivanova, Natalia; Kyrpides, Nikos; Reeve, Wayne

    2015-01-01

    Bradyrhizobium sp. WSM1253 is a novel N2-fixing bacterium isolated from a root nodule of the herbaceous annual legume Ornithopus compressus that was growing on the Greek Island of Sifnos. WSM1253 emerged as a strain of interest in an Australian program that was selecting inoculant quality bradyrhizobial strains for inoculation of Mediterranean species of lupins (Lupinus angustifolius, L. princei, L. atlanticus, L. pilosus). In this report we describe, for the first time, the genome sequence information and annotation of this legume microsymbiont. The 8,719,808 bp genome has a G + C content of 63.09 % with 71 contigs arranged into two scaffolds. The assembled genome contains 8,432 protein-coding genes, 66 RNA genes and a single rRNA operon. This improved-high-quality draft rhizobial genome is one of 20 sequenced through a DOE Joint Genome Institute 2010 Community Sequencing Project.

  13. Draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite strain recommended for cowpea inoculation in Brazil.

    Science.gov (United States)

    Simões-Araújo, Jean Luiz; Leite, Jakson; Passos, Samuel Ribeiro; Xavier, Gustavo Ribeiro; Rumjanek, Norma Gouvêa; Zilli, Jerri Édson

    The strain BR 3267 is a nitrogen-fixing symbiotic bacteria isolated from soil of semi-arid area of Brazilian Northeast using cowpea as the trap plant. This strain is used as commercial inoculant for cowpea and presents high efficient in nitrogen fixation as consequence of its adaptation potential to semi-arid conditions. We report here the draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite bacterium used as inoculant for cowpea. Whole genome sequencing of BR 3267 using Illumina MiSeq sequencing technology has 55 scaffolds with a total genome size of 7,904,309bp and C+G 63%. Annotation was added by the RAST prokaryotic genome annotation service and has shown 7314 coding sequences and 52 RNA genes.

  14. Draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite strain recommended for cowpea inoculation in Brazil

    Directory of Open Access Journals (Sweden)

    Jean Luiz Simões-Araújo

    Full Text Available Abstract The strain BR 3267 is a nitrogen-fixing symbiotic bacteria isolated from soil of semi-arid area of Brazilian Northeast using cowpea as the trap plant. This strain is used as commercial inoculant for cowpea and presents high efficient in nitrogen fixation as consequence of its adaptation potential to semi-arid conditions. We report here the draft genome sequence of Bradyrhizobium sp. strain BR 3267, an elite bacterium used as inoculant for cowpea. Whole genome sequencing of BR 3267 using Illumina MiSeq sequencing technology has 55 scaffolds with a total genome size of 7,904,309 bp and C+G 63%. Annotation was added by the RAST prokaryotic genome annotation service and has shown 7314 coding sequences and 52 RNA genes.

  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. Ultrastructural and immunocytochemical studies on the MLS-type basal apparatus in the sperm of Lygodium japonicum (pteridophyta)

    OpenAIRE

    Sakaushi, Shinji

    2004-01-01

    Spiral formation of the multiflagellated sperm in Lygodium japonicum(pteridophyte) was ultrastructurally and biochemically analyzed to consider evolution of the basal apparatus in the archegoniates.Differentiation of the spermatid (spermiogenesis) was investigated in detail by electron microscopy, immunofluorescence and immunoelectron microscopy using anti-tubulin and anti-actin antibodies. At the mid stage of spermiogenesis, the MLS, a characteristic structure in the archegoniates and the C...

  17. Effect of photoperiod change on chronobiology of cercarial emergence of Schistosoma japonicum derived from hilly and marshy regions of China.

    Science.gov (United States)

    Wang, Su-Rong; Zhu, Yuan-Jian; Ge, Qing-Peng; Yang, Meng-Jia; Huang, Ji-Lei; Huang, Wen-Qiao; Zhuge, Hong-Xiang; Lu, Da-Bing

    2015-12-01

    The chronobiology of cercarial emergence appeared to be a genetically controlled behavior, adapted to definitive host species, for schistosome. However, a few physiological and ecological factors, for example the change of photoperiod, were reported to affect the rhythmic emergence of cercariae. Therefore, the effect of photoperiod change on cercarial emergence of two Schistosoma japonicum isolates, the hilly and the marshland, was investigated. Four shedding experiments each under a different photoperiod were conducted. Under a natural photoperiod, two distinct shedding modes, one from the hilly region and one from the marshland, were observed. Under a reversed photoperiod, the regular pattern (i.e. under a natural photoperiod) of S. japonicum cercarial emergence was reversed for the marshland isolate and disappeared for the hilly isolate. With an input of a 2 h darkness from 7am to 9am, the cercarial emergence peak were delayed for the two isolates; whereas with an input of a 2 h darkness from 5pm to 7pm, neither effect on the cercarial emergence rhythm was observed. The total cercariae emerged for both parasite isolates varied with a different photoperiod. The results indicate that the change of photoperiod could affect the chronobiology of S japonicum cercarial emergence.

  18. Suppression of the Insulin Receptors in Adult Schistosoma japonicum Impacts on Parasite Growth and Development: Further Evidence of Vaccine Potential.

    Science.gov (United States)

    You, Hong; Gobert, Geoffrey N; Cai, Pengfei; Mou, Rong; Nawaratna, Sujeevi; Fang, Guofu; Villinger, Francois; McManus, Donald P

    2015-05-01

    To further investigate the importance of insulin signaling in the growth, development, sexual maturation and egg production of adult schistosomes, we have focused attention on the insulin receptors (SjIRs) of Schistosoma japonicum, which we have previously cloned and partially characterised. We now show, by Biolayer Interferometry, that human insulin can bind the L1 subdomain (insulin binding domain) of recombinant (r)SjIR1 and rSjIR2 (designated SjLD1 and SjLD2) produced using the Drosophila S2 protein expression system. We have then used RNA interference (RNAi) to knock down the expression of the SjIRs in adult S. japonicum in vitro and show that, in addition to their reduced transcription, the transcript levels of other important downstream genes within the insulin pathway, associated with glucose metabolism and schistosome fecundity, were also impacted substantially. Further, a significant decrease in glucose uptake was observed in the SjIR-knockdown worms compared with luciferase controls. In vaccine/challenge experiments, we found that rSjLD1 and rSjLD2 depressed female growth, intestinal granuloma density and faecal egg production in S. japonicum in mice presented with a low dose challenge infection. These data re-emphasize the potential of the SjIRs as veterinary transmission blocking vaccine candidates against zoonotic schistosomiasis japonica in China and the Philippines.

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

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    Akio Tani

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

  20. Isolation and characterization of symbiotic mutants of bradyrhizobium sp. (Arachis) strain NC92: mutants with host-specific defects in nodulation and nitrogen fixation.

    OpenAIRE

    Wilson, K. J.; Anjaiah, V; Nambiar, P T; Ausubel, F M

    1987-01-01

    Random transposon Tn5 mutagenesis of Bradyrhizobium sp. (Arachis) strain NC92, a member of the cowpea cross-inoculation group, was carried out, and kanamycin-resistant transconjugants were tested for their symbiotic phenotype on three host plants: groundnut, siratro, and pigeonpea. Two nodulation (Nod- phenotype) mutants were isolated. One is unable to nodulate all three hosts and appears to contain an insertion in one of the common nodulation genes (nodABCD); the other is a host-specific nod...

  1. 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.

  2. Diversidade fenotípica e eficiência simbiótica de estirpes de Bradyrhizobium spp. de solos da Amazônia Phenotypic diversity and symbiotic efficiency of Bradyrhizobium spp. strains from Amazonian soils

    Directory of Open Access Journals (Sweden)

    Adriana Silva Lima

    2005-11-01

    Full Text Available Este trabalho teve como objetivo avaliar a diversidade fenotípica e a eficiência simbiótica de estirpes de Bradyrhizobium, isoladas de solos da Amazônia, sob diferentes sistemas de uso da terra (monocultura, capoeira, pastagem, floresta e sistema agroflorestal. A análise dos perfis de proteína total de 46 estirpes, obtidos por eletroforese em gel de poliacrilamida (SDS-PAGE, mostrou grande diversidade, tendo formado 11 grupos com similaridade acima de 80%. Apenas um dos grupos continha a estirpe referência de B. elkanii: BR29, recomendada como inoculante para soja. Vinte e duas estirpes testadas em vasos de Leonard, com caupi (Vigna unguiculata (L. Walp., induziram à produção de matéria seca e ao acúmulo de nitrogênio, na parte aérea da planta, e à eficiência relativa superiores aos da testemunha (sem N e sem inoculação. Entre as estirpes testadas, 13 induziram à produção de matéria seca e à eficiência relativa similares às da testemunha nitrogenada (com N, sem inoculação; cinco estirpes induziram a acúmulo de N superior ao da testemunha nitrogenada. Essas populações nativas são constituídas por grande diversidade de estirpes, com eficiência simbiótica variável, algumas das quais podem ser recomendadas para testes de eficiência agronômica.This work aimed to evaluate the phenotypic diversity and symbiotic efficiency of Bradyrhizobium strains isolated from Amazonian soils, under different land use systems (crop, fallow, pasture, forest and agroforestry system. Total protein profiles obtained by polyacrylamide gel electrophoresis (SDS-PAGE of 46 strains showed great diversity, while 11 groups presented similarity above 80%. One of these groups contained the reference strain of B. elkanii: BR29, recommended as soybean inoculant. Twenty-two strains, tested in Leonard jars for symbiotic efficiency with Vigna unguiculata (L. Walp. (cowpea produced shoot dry matter, N-content, and relative efficiency higher than the

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

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    D. L. Miguel

    2001-12-01

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

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

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    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.

  5. 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.

  6. 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.

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

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    Lin Chen

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

  8. BEBERAPA FAKTOR YANG BERHUBUNGAN DENGAN PENULARAN Schistosoma japonicum DI DATARAN TINGGI NAPU KABUPATEN POSO SULAWESI TENGAH

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    Rosmini Rosmini

    2012-07-01

    Full Text Available Abstract. The transmission of schistosomiasis is related with human behavior or habit factors in the schistosomiasis endemic area. The Purpose of this study was to identify several factors in relationship with schistosomiasis in Napu Highland. The Study was observational with cross sectional design. Data collection were coducted stool survey and interviewes in among 196 respondents using simple random sampling. Stool samples were examined by the Kato-Katz method and interviewed was used  questionnaire to identify several factors in realtionship with schistosomiasis transmission. Out of 6 community behaviours suspected to contribute to infection of schistosomiasis, 5 were related with schistosomiasis, i. e. the habit of taking a bath and washing in the river (p=<0.001, always going to the focus area (p=0.04,utilizing water resources from well (p=0.001, washing legs or hand in the river(p=<0.001 and swimming (p=0.007. Schistosomiasis infection was related to the community behaviour of bathing and washing in the river, always going to the focus area, utilizing water resources from well, washing legs or hand in the river and swimming.Key word: Schistosomiasis, Schistosoma japonicum, behaviour, Napu Highland.Central Sulawesi

  9. 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.

  10. 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.

  11. Bradyrhizobium BclA Is a Peptide Transporter Required for Bacterial Differentiation in Symbiosis with Aeschynomene Legumes.

    Science.gov (United States)

    Guefrachi, Ibtissem; Pierre, Olivier; Timchenko, Tatiana; Alunni, Benoît; Barrière, Quentin; Czernic, Pierre; Villaécija-Aguilar, José-Antonio; Verly, Camille; Bourge, Mickaël; Fardoux, Joël; Mars, Mohamed; Kondorosi, Eva; Giraud, Eric; Mergaert, Peter

    2015-11-01

    Nodules of legume plants are highly integrated symbiotic systems shaped by millions of years of evolution. They harbor nitrogen-fixing rhizobium bacteria called bacteroids. Several legume species produce peptides called nodule-specific cysteine-rich (NCR) peptides in the symbiotic nodule cells which house the bacteroids. NCR peptides are related to antimicrobial peptides of innate immunity. They induce the endosymbionts into a differentiated, enlarged, and polyploid state. The bacterial symbionts, on their side, evolved functions for the response to the NCR peptides. Here, we identified the bclA gene of Bradyrhizobium sp. strains ORS278 and ORS285, which is required for the formation of differentiated and functional bacteroids in the nodules of the NCR peptide-producing Aeschynomene legumes. The BclA ABC transporter promotes the import of NCR peptides and provides protection against the antimicrobial activity of these peptides. Moreover, BclA can complement the role of the related BacA transporter of Sinorhizobium meliloti, which has a similar symbiotic function in the interaction with Medicago legumes.

  12. 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

    2016-09-01

    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.

  13. 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

  14. 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-

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

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    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.

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

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    Santana Clara

    2009-10-01

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

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

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    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.

  20. Compatibility among fungicide treatments on soybean seeds through film coating and inoculation with Bradyrhizobium strains = Compatibilidade entre tratamento fungicida de sementes de soja via peliculização e inoculação com estirpes de Bradyrhizobium

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    Carla Massimo Caldeira

    2010-10-01

    Full Text Available This research evaluated the development of soybean plants grown in agreenhouse, from seeds that were treated with fungicide, film coated and inoculated with Bradyrhizobium. Seeds from the soybean cultivar ‘Vencedora’ were submitted to treatments disposed in a factorial 5 x 3 x 2 design. The seeds were treated with Derosal Plus. with polymer; Tegran. with polymer; Derosal Plus. without polymer, Tegran. without polymer; and without fungicide and without polymer. After the chemicaltreatment, the seeds were inoculated with peat or liquid inoculants, and without inoculant. The chemical treatments were used either previous to or after the six months of storage. Seed inoculation was performed with CPAC15 and BR29 strains, immediately before sowing in Leonard pots (two seedlings per pot. The plants at R2 stage wereevaluated through the number and dry matter weight of nodules, the shoot dry matter weight, and the chlorophyll content. The results indicated that the treatments with Derosal Plus or Tegran-associated or not with the polymer, and independent of the application time-did not affect nodule establishment and development, or nitrogen fixation when the seeds were inoculated. Liquid and peat inoculants applied to the seeds were equally efficient in the formation of nodules in soybean plants; however, the liquid inoculant promoted higher shoot dry matter weight.Esta pesquisa foi conduzida com o objetivo de avaliar o desenvolvimento em casa-de-vegetação da cultura da soja, proveniente de sementes submetidas ao tratamento fungicida, peliculização e inoculação com Bradyrhizobium. Sementes da cultivar Vencedora foram submetidas a tratamentos dispostos em fatorial 5 x 3 x 2, sendo: sem fungicida e sem polímero, Derosal Plus. com polímero, Tegran. com polímero, Derosal Plus. sem polímero e Tegran. sem polímero; inoculante líquido, turfoso ou não-inoculadas; antes ou após seis meses de armazenamento. A inoculação das sementes foi

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

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    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

  2. 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

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    Alexandre Barberi

    2004-04-01

    Full Text Available 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 utilizadas estirpes tolerantes à acidez do solo ou induzir tolerância nas estirpes já utilizadas. Em alguns trabalhos, tem-se demonstrado indução de tolerância à acidez quando a bactéria é previamente exposta a pH ácido. Assim, objetivou-se com este trabalho verificar o crescimento de Bradyrhizobium elkanii estirpe Br 29 em meios de cultura sob diferentes condições de pH inicial. Foram realizados dois experimentos com dois meios de cultivo diferentes, em que se variou o pH inicial (6,8; 6,0; 5,5. Foram avaliados: o crescimento da estirpe, pela contagem do número de unidades formadoras de colônia (UFC e por densidade ótica, assim como as mudanças do pH nos meios. As curvas de crescimento obtidas variaram de acordo com o pH inicial e com a composição do meio de cultura. Com base no número máximo de UFC.mL-1, para o meio Lorda & Balatti modificado, o melhor pH foi 6,0 e, no meio Lopreto, o melhor pH foi 5,5. O melhor crescimento em pH mais baixo (5,5 está associado à maior produção de exopolissacarídeo.Currently soybean is the most economically important legume crop in Brazil. Due to its profitability soybean is being widely spread out all over the national territory. Some assays have shown low response of this crop to liming in acid Brazilian soil, under no-till age systems. Acidity is one of the factors that limit the efficiency of the symbiosis rhizobia - legumes. This problem could be solved if acid tolerant strains were

  3. NifDK clusters located on the chromosome and megaplasmid of Bradyrhizobium sp strain DOA9 contribute differently to nitrogenase activity during symbiosis and free-living growth

    OpenAIRE

    Wongdee, J.; Songwattana, P.; Nouwen, Nico; Noisangiam, R.; Fardoux, Joël; Chaintreuil, Clémence; Teaumroong, N.; Tittabutr, P.; Giraud, Eric

    2016-01-01

    Bradyrhizobium sp. strain DOA9 contains two copies of the nifDK genes, nifDKc, located on the chromosome, and nifDKp, located on a symbiotic megaplasmid. Unlike most rhizobia, this bacterium displays nitrogenase activity under both free-living and symbiotic conditions. Transcriptional analysis using gusA reporter strains showed that both nifDK operons were highly expressed under symbiosis, whereas nifDKc was the most abundantly expressed under free-living conditions. During free-living growth...

  4. 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.

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

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

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    Zhang Donghui

    2011-06-01

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

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

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    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.

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

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

    2014-02-01

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

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

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    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

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

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    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.

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

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    Nathanael Delmotte

    2014-02-01

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

  13. 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

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

    Science.gov (United States)

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

    2017-04-01

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

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

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    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.

  16. (±)-Japonones A and B, two pairs of new enantiomers with anti-KSHV activities from Hypericum japonicum

    Science.gov (United States)

    Hu, Linzhen; Zhu, Hucheng; Li, Lei; Huang, Jinfeng; Sun, Weiguang; Liu, Junjun; Li, Hua; Luo, Zengwei; Wang, Jianping; Xue, Yongbo; Zhang, Yu; Zhang, Yonghui

    2016-06-01

    Two pairs of new enantiomers with unusual 5,5-spiroketal cores, termed (±)-japonones A and B [(±)-1 and (±)-2], were obtained from Hypericum japonicum Thunb. The absolute configurations of (±)-1 and (±)-2 were characterized by extensive analyses of spectroscopic data and calculated electronic circular dichroism (ECD) spectra, the application of modified Mosher’s methods, and the assistance of quantum chemical predictions (QCP) of 13C NMR chemical shifts. Among these metabolites, (+)-1 exhibited some inhibitory activity on Kaposi’s sarcoma associated herpesvirus (KSHV). Virtual screening of (±)-1 and (±)-2 were conducted using the Surflex-Dock module in the Sybyl software, and (+)-1 exhibited ability to bind with ERK to form key interactions with residues Lys52, Pro56, Ile101, Asp165, Gly167 and Val99.

  17. Fluorescence studies with malate dehydrogenase from rhizobium japonicum 3I1B-143 bacteroids: a two-tryptophan containing protein

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    Ghiron, Camillo A.; Eftink, Maurice R.; Waters, James K.; Emerich, David W.

    1990-05-01

    A number of fluorescence studies, both of trp residues and bound NADH, have been reported for porcine MDH. The large number of trp residues (6) complicates the interpretation of some studies. To circumvent this we have performed studies with a two tryptophan (per subunit) MDH from Rhizobium japonicum 311B-143 bacteroids. We have performed phase/modulation fluorescence lifetime measurements, as a function of temperature and added quencher KI, in order to resolved the 1.3 ns (blue) and 6.6 ns (red) contributions from the two classes of trp residues. Anisotropy decay studies have also been performed. The binding of NADH dynamically quenches the fluorescence of both tip residues, but, unlike mammalian cytoplasmic and mitochondrial MDH, there is not a large enhancement in fluorescence of bound NADH upon forming a ternary complex with either tartronic acid or D-malonate.

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

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    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.

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

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    Liu Feng

    2010-11-01

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

  20. 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.

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

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    Jose Ma M Angeles

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

  2. An "in-depth" description of the small non-coding RNA population of Schistosoma japonicum schistosomulum.

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    Zhangxun Wang

    Full Text Available Parasitic flatworms of the genus Schistosoma are the causative agents of schistosomiasis, which afflicts more than 200 million people yearly in tropical regions of South America, Asia and Africa. A promising approach to the control of this and many other diseases involves the application of our understanding of small non-coding RNA function to the design of safe and effective means of treatment. In a previous study, we identified five conserved miRNAs from the adult stage of Schistosoma japonicum. Here, we applied Illumina Solexa high-throughput sequencing methods (deep sequencing to investigate the small RNAs expressed in S. japonicum schistosomulum (3 weeks post-infection. This has allowed us to examine over four million sequence reads including both frequently and infrequently represented members of the RNA population. Thus we have identified 20 conserved miRNA families that have orthologs in well-studied model organisms and 16 miRNA that appear to be specific to Schistosoma. We have also observed minor amounts of heterogeneity in both 3' and 5' terminal positions of some miRNA as well as RNA fragments resulting from the processing of miRNA precursor. An investigation of the genomic arrangement of the 36 identified miRNA revealed that seven were tightly linked in two clusters. We also identified members of the small RNA population whose structure indicates that they are part of an endogenously derived RNA silencing pathway, as evidenced by their extensive complementarities with retrotransposon and retrovirus-related Pol polyprotein from transposon.

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

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    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

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

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    Li Yuesheng

    1993-08-01

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

  5. Comparison of the 'Ca Liberibacter asiaticus' genome adapted for an intracellular lifestyle with other members of the rhizobiales

    Science.gov (United States)

    An intracellular plant pathogen ‘Ca. Liberibacter asiaticus,’ a member of the Rhizobiales, is related to Sinorhizobium meliloti, Bradyrhizobium japonicum, Agrobacterium tumefaciens and Bartonella henselae, an intracellular mammalian pathogen. Whole chromosome comparisons identified at least 52 clust...

  6. Conservation of gene order and content in the circular chromosomes of 'Candidatus Liberibacter' asiaticus and other rhizbiales

    Science.gov (United States)

    The intracellular plant pathogen ‘Ca. Liberibacter asiaticus’ is a member of the Rhizobiales, as are the nitrogen fixing Sinorhizobium meliloti and Bradyrhizobium japonicum, the plant pathogen Agrobacterium tumefaciens and the intracellular mammalian pathogen Bartonella henselae. Whole genome compar...

  7. 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.

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

    Science.gov (United States)

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

    2015-01-01

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

  9. Surviving and thriving in terms of symbiotic performance of antibiotic and phage-resistant mutants of Bradyrhizobium of soybean [Glycine max (L.) Merrill].

    Science.gov (United States)

    Anand, Akhil; Jaiswal, Sanjay Kumar; Dhar, Banshi; Vaishampayan, Akhouri

    2012-10-01

    Rhizobial inoculation plays an important role in yielding enhancement of soybean, but it is frequently disturbed by competition with bacterial population present in the soil. Identification of potential indigenous rhizobia as competitive inoculants for efficient nodulation and N(2)-fixation of soybean was assessed under laboratory and field conditions. Two indigenous bradyrhizobial isolates (MPSR033 and MPSR220) and its derived different antibiotic (streptomycin and gentamicin) and phage (RT5 and RT6)-resistant mutant strains were used for competition study. Nodulation occupancy between parent and mutant strains was compared on soybean cultivar JS335 under exotic condition. Strain MPSR033 Sm(r) V(r) was found highly competitive for nodule occupancy in all treatment combinations. On the basis of laboratory experiments four indigenous strains (MPSR033, MPSR033 Sm(r), MPSR033 Sm(r) V(r), MPSR220) were selected for their symbiotic performance along with two exotic strains (USDA123 and USDA94) on two soybean cultivars under field conditions. A significant symbiotic interaction between Bradyrhizobium strains and soybean cultivar was observed. Strain MPSR033 Sm(r) V(r) was found superior among the rhizobial treatments in seed yield production with both cultivars. The 16S rRNA region sequence analysis of the indigenous strains showed close relationship with Bradyrhizobium yuanmingense strain. These findings widen out the usefulness of antibiotic-resistance marked phage-resistant bradyrhizobial strains in interactive mode for studying their symbiotic effectiveness with host plant, and open the way to study the mechanism of contact-dependent growth inhibition in rhizobia.

  10. 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.

  11. 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.

  12. Confocal laser scanning microscopic observation on adult Schistosoma japonicum harbored in mice following treatment with single-dose mefloquine.

    Science.gov (United States)

    Xiao, Shu-Hua; Sun, Jun; Xue, Jian

    2012-06-01

    The aim of the present study is to assess the mefloquine-induced alteration of adult Schistosoma japonicum using confocal laser scanning microscopy (CLSM). Eight out of ten mice infected with 60-80 S. japonicum cercariae for 35 days were treated orally with mefloquine at a single dose of 400 mg/kg. Four groups of two mice were killed at 24 h and 3, 7, and 14 days post-treatment, and schistosomes were collected by perfusion from the liver and mesenteric veins, fixed in 70% alcohol, stained with acid carmine, and examined by CLSM. Worms obtained from untreated mice served as controls. Twenty-four hours post-treatment, focal tegument of adult male and female worms, which composed of fine and short villus-like materials, became thicker and longer, or disorder arrangement, while the musculatures beneath the tegument revealed in focal and irregular swelling with various degrees. In the gut of male and female schistosomes, severe dilatation accompanied by swelling, collapse, and peeling of gut mucosa was universal. In the reproductive organs, no apparent alteration in the testis structure of male worms was seen, while in female worms, slight damage to the ovary included loose arrangement of mature ovary cells accompanied by some of them degenerated and collapsed. As to vitelline glands, severe damage, such as swelling, indistinction, fusion or collapse of vitelline cells, and apparent swelling of parenchymal tissues in vitelline gland lobules, was seen. Meanwhile, abnormal ova emerged in the uterus at this time point. Three to 7 days post-treatment, the damage to the worms aggravated either in extent or in severity along with time. In some focally swollen worm body, the parenchymal tissues revealed in severe swelling. In addition, a large piece of degenerated and necrotic parenchymal tissues emerged closed to the severe destructed oral or ventral sucker. In the gut of male and female worms, the major alterations manifested by focal collapse or peeling of mucosa, and

  13. Further studies on mefloquine and praziquantel alone or interaction of both drugs against Schistosoma japonicum in vitro.

    Science.gov (United States)

    Xiao, Shu-hua; Xue, Jian; Zhang, Hao-bing

    2012-03-01

    The aim of the present study is to further understand and analyze the interaction of mefloquine with praziquantel against adult Schistosoma japonicum in vitro. Mice infected with S. japonicum cercariae for 35-37 days were sacrificed, and adult schistosomes were collected by perfusion. Schistosomes were placed to each of 12 wells of a Falcon plate and maintained in RPMI 1640 supplemented by 10% calf serum. For determination of 50% and 95% lethal concentration (LC50 and LC95) of the two drugs in vitro, schistosomes were exposed to mefloquine at concentrations of 1, 2, 3, 4, 5, 6, 7, and 10 μg/mL or praziquantel at concentrations of 0.001, 0.01, 0.05, 0.1, 0.2, 0.5, 1, 10, and 30 μg/mL. The plate was incubated at 37°C in 95% air + 5% CO₂ for 72 h. According to the half-life of oral mefloquine and praziquantel in mice, mefloquine combined with praziquantel simultaneously, mefloquine administered within 1 h after praziquantel and praziquantel administered within 17 h after mefloquine were used to evaluate the effect of mefloquine in combination with praziquantel against S. japonicum in vitro. The results showed that the LC50 and LC95 of mefloquine calculated by the Bliss method were 6.17 μg/mL (95% confidence limits, 5.84-6.517 μg/mL) and 8.703 μg/mL (95% confidence limits, 7.632-9.797 μg/mL), respectively. As to praziquantel, no worm death was seen when schistosomes were exposed to praziquantel at concentrations of 0.005-0.2 μg/mL for 72 h. While in the worms exposed to praziquantel 1, 10, and 30 μg/mL, strong spasmodic contractions of the worm body and vesiculation along the worm surface were observed, but 48-75% of the schistosomes survived the exposure in 72-h incubation. Meanwhile, the number of dead worms that emerged in each group was not proportion to the increasing concentrations. Therefore, it is not appropriate to calculate the LC50 and LC95 of praziquantel. For evaluation of the interaction with the two drugs, praziquantel 0.1 or 0.2

  14. 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.

  15. 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.

  16. Determination of Flavonoids in Different Parts and Different Harvesting Seasons of Hypericum japonicum%不同采收季节及部位田基黄药材中黄酮类成分的含量测定

    Institute of Scientific and Technical Information of China (English)

    席仲洪; 虞金宝; 吕武清; 李晶; 李才堂

    2009-01-01

    目的:测定不同采收季节及部位田基黄药材中黄酮类成分的含量.方法:采用比色法测定田基黄药材中总黄酮的含量;采用高效液相色谱(HPLC)法测定田基黄药材中槲皮素、芦丁、异鼠李素的含量.结果:不同采收季节田基黄药材中总黄酮的含量无显著差异;广西产田基黄药材中不含异鼠李素,但槲皮素、芦丁含量较高.田基黄药材不同部位中总黄酮含量以种子和叶高,根中低,且根中几乎不含异鼠李素、槲皮素、芦丁.结论:由于春季田基黄药材植株矮小、产量低,因此应以秋季采收为佳.规定田基黄药材的药用部位为全草是科学的.%OBJECTIVE: To determine the contents of flavonoids in different part and different harvesting seasons of Hypericum japonicum. METHODS: The total content of flavonoids in H. japonicum was determined by colorimetry and the contents of quercetin, rutin and isorhamnetin in H, japonicum were determined by HPLC. RESULTS: The H. japonicum harvested in different seasons showed no significant difference in terms of the contents of total flavonoids, H. japonicum from Guangxi province contained no isorhamnetin but which was rich in the contents of quercetin and rutin. In H. japonicum, the content of total flavonoids was rich in the seeds and leaves but poor in the roots, and hardly had isorhamnetin, quercetin and rutin been detected in the roots. CONCLUSION: H. japonicum harvested in the spring season is low and small in size with low - yield; therefore, fall is its optimal harvesting season. Furthermore, the specification in Chinese Pharmacopeia that the medicinal part of H. japonicum is the entire herb has been proved to be scientific.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. Growth Rate of and Gene Expression in Bradyrhizobium diazoefficiens USDA110 due to a Mutation in blr7984, a TetR Family Transcriptional Regulator Gene.

    Science.gov (United States)

    Ohkama-Ohtsu, Naoko; Honma, Haruna; Nakagome, Mariko; Nagata, Maki; Yamaya-Ito, Hiroko; Sano, Yoshiaki; Hiraoka, Norina; Ikemi, Takaaki; Suzuki, Akihiro; Okazaki, Shin; Minamisawa, Kiwamu; Yokoyama, Tadashi

    2016-09-29

    Previous transcriptome analyses have suggested that a gene cluster including a transcriptional regulator (blr7984) of the tetracycline repressor family was markedly down-regulated in symbiosis. Since blr7984 is annotated to be the transcriptional repressor, we hypothesized that it is involved in the repression of genes in the genomic cluster including blr7984 in symbiotic bacteroids. In order to examine the function and involvement of the blr7984 gene in differentiation into bacteroids, we compared the free-living growth/symbiotic phenotype and gene expression between a blr7984-knockout mutant and the wild-type strain of Bradyrhizobium diazoefficiens USDA110. The mutant transiently increased the cell growth rate under free-living conditions and nodule numbers over those with the wild-type strain USDA110. The expression of three genes adjacent to the disrupted blr7984 gene was strongly up-regulated in the mutant in free-living and symbiotic cells. The mutant also induced the expression of genes for glutathione S-transferase, cytochrome c oxidases, ABC transporters, PTS sugar transport systems, and flagella synthesis under free-living conditions. bll7983 encoding glutathione S-transferase was up-regulated the most by the blr7984 disruption. Since redox regulation by glutathione is known to be involved in cell division in prokaryotes and eukaryotes, the strong expression of glutathione S-transferase encoded by the bll7983 gene may have caused redox changes in mutant cells, which resulted in higher rates of cell division.

  2. Performance of a dipstick dye immunoassay for rapid screening of Schistosoma japonicum infection in areas of low endemicity

    Directory of Open Access Journals (Sweden)

    Peeling Rosanna W

    2011-05-01

    Full Text Available Abstract Background The dipstick dye immunoassay (DDIA, recently commercially available in the People's Republic of China (P.R. China, is a rapid and simple test to detect human antibodies against Schistosoma Japonicum. Its performance and utility for screening schistosome infection in low endemic areas is little known. We therefore carried out a cross-sectional survey in seven villages with low endemicity of schistosomiasis in P.R. China and assessed the performance and utility of DDIA for diagnosis of schistosomiasis. Stool samples were collected and examined by the Kato-Katz method and the miracidium hatching technique. Serum samples, separated from whole blood of participants, were tested by DDIA. Results 6285 individuals aged 6-65 years old participated in this study, with a prevalence of schistosomiasis of 4.20%. Using stool examination as a gold reference standard, DDIA performed with a high overall sensitivity of 91.29% (95% CI: 87.89-94.69% and also a high negative predictive value, with a mean value of 99.29% (95% CI: 98.99-99.58%. The specificity of DDIA was only moderate (53.08%, 95% CI: 51.82-54.34%. Multivariate analysis indicated that age, occupation and history of schistosome infection were significantly associated with the false positive results of DDIA. Conclusions DDIA is a sensitive, rapid, simple and portable diagnostic assay and can be used as a primary approach for screening schistosome infection in areas of low endemicity. However, more sensitive and specific confirmatory assays need to be developed and combined with DDIA for targeting chemotherapy accurately.

  3. Genomic organization and genomic structural rearrangements of Sphingobium japonicum UT26, an archetypal γ-hexachlorocyclohexane-degrading bacterium.

    Science.gov (United States)

    Nagata, Yuji; Natsui, Shunsuke; Endo, Ryo; Ohtsubo, Yoshiyuki; Ichikawa, Natsuko; Ankai, Akiho; Oguchi, Akio; Fukui, Shigehiro; Fujita, Nobuyuki; Tsuda, Masataka

    2011-12-10

    The complete genome sequencing of a γ-hexachlorocyclohexane-degrading strain, Sphingobium japonicum UT26, revealed that the genome consists of two circular chromosomes [with sizes of 3.5 Mb (Chr1) and 682kb (Chr2)], a 191-kb large plasmid (pCHQ1), and two small plasmids with sizes of 32 and 5kb. The lin genes are dispersed on Chr1, Chr2, and pCHQ1. Comparison of the UT26 genome with those of other sphingomonad strains demonstrated that the "specific"lin genes for conversion of γ-HCH to β-ketoadipate (linA, linB, linC, linRED, and linF) are located on the DNA regions unique to the UT26 genome, suggesting the acquisition of these lin genes by horizontal transfer events. On the other hand, linGHIJ and linKLMN are located on the regions conserved in the genomes of sphingomonads, suggesting that the linGHIJ-encoded β-ketoadipate pathway and the LinKLMN-type ABC transporter system are involved in core functions of sphingomonads. Based on these results, we propose a hypothesis that UT26 was created by recruiting the specific lin genes into a strain having core functions of sphingomonads. Most of the specific lin genes in UT26 are associated with IS6100. Our analysis of spontaneous linA-, linC-, and linRED-deletion mutants of UT26 revealed the involvement of IS6100 in their deduced genome rearrangements. These facts strongly suggest that IS6100 plays important roles both in the dissemination of the specific lin genes and in the genome rearrangements.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. CARACTERIZACIÓN QUÍMICA DEL GRANO MOLIDO Y LA TORTA DE SOYA DESGRASADA (Glycine max, Merr; VAR. INCA SOY 24) QUE SE EMPLEAN EN EL MEDIO DE CULTIVO PARA Bradyrhizobium elkanii

    OpenAIRE

    Aida T. Rodríguez; María C. Nápoles; M. A. Ramírez; Yamilet Gutiérrez; Daimy Costales

    2004-01-01

    Algunos derivados de la soya se han utilizado como medio de cultivo para Bradyrhizobium, demostrándose su efecto positivo sobre la dinámica de crecimiento de estos microorganismos, así como su poder inductor sobre la expresión de los genes de nodulación. El presente trabajo se llevó a cabo en el Departamento de Fisiología y Bioquímica Vegetal, perteneciente al Instituto Nacional de Ciencias Agrícolas (INCA), donde se realizó la caracterización química del grano molido y la torta desgrasada de...

  9. 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

  10. 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.

  11. Specific anti-glycan antibodies are sustained during and after parasite clearance in Schistosoma japonicum-infected rhesus macaques

    Science.gov (United States)

    Yang, Y. Y. Michelle; Li, Xiao Hong; Brzezicka, Katarzyna; Reichardt, Niels-Christian; Wilson, R. Alan; van Diepen, Angela

    2017-01-01

    Background Human immunity to Schistosoma infection requires many years of exposure, and multiple infections and treatments to develop. Unlike humans, rhesus macaques clear an established schistosome infection naturally at the same time acquiring immunity towards re-infection. In macaques, schistosome egg production decreases after 8 weeks post-infection and by week 22, physiological impairment of the worm caused by unclarified antibody-mediated processes is observed. Since strong antibody responses have been observed against schistosome glycan antigens in human and animal infections, we here investigate if anti-glycan antibodies are associated with immunity against schistosome infections in macaques. Methods We used a microarray containing a large repertoire of glycoprotein- and glycolipid-derived glycans from different schistosome life stages to analyse anti-glycan serum IgG and IgM from S. japonicum-infected macaques during the course of infection and self-cure. We also used an in vitro schistosomula assay to investigate whether macaque sera containing anti-glycan antibodies can kill schistosomula. Conclusions/significance Antibody responses towards schistosome glycans at week 4 post-infection were dominated by IgM while IgG was high at week 8. The profound increase in IgG was observed mainly for antibodies towards a large subset of glycans that contain (multi-)fucosylated terminal GalNAcβ1-4GlcNAc (LDN), and Galβ1-4(Fucα1–3)GlcNAc (LeX) motifs. In general, glycans with a higher degree of fucosylation gave rise to stronger antibody responses than non-fucosylated glycans. Interestingly, even though many IgG and IgM responses had declined by week 22 post-infection, IgG towards O-glycans with highly fucosylated LDN motifs remained. When incubating macaque serum with schistosomula in vitro, schistosomula death was positively correlated with the duration of infection of macaques; macaque serum taken 22 weeks post-infection caused most schistosomula to die

  12. 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.

  13. 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)

  14. 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.

  15. 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.

  16. 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.

  17. Evaluation of banked urine samples for the detection of circulating anodic and cathodic antigens in Schistosoma mekongi and S. japonicum infections: a proof-of-concept study.

    Science.gov (United States)

    van Dam, Govert J; Odermatt, Peter; Acosta, Luz; Bergquist, Robert; de Dood, Claudia J; Kornelis, Dieuwke; Muth, Sinuon; Utzinger, Jürg; Corstjens, Paul L A M

    2015-01-01

    In Asia, Schistosoma japonicum is the predominant schistosome species, while Schistosoma mekongi is confined to limited foci in Cambodia and Lao People's Democratic Republic. While the People's Republic of China has been successful in controlling schistosomiasis, the disease remains a major public health issue in other areas. In order to prioritise intervention areas, not only accurate diagnosis is important but also other factors, such as practicality, time-efficiency and cost-effectiveness, since they strongly influence the success of control programmes. To evaluate the highly specific urine-based assays for the schistosome circulating cathodic antigen (CCA) and the circulating anodic antigen (CAA), banked urine samples from Cambodia (n=106) and the Philippines (n=43) were examined by the upconverted phosphor lateral flow (UCP-LF) CAA assay and the point-of-care (POC)-CCA urine assay. Based on 250 μl urine samples, UCP-LF CAA sensitivity outcomes surpassed a single stool examination by the Kato-Katz technique. The banked urine samples in the current study did not allow the evaluation of larger volumes, which conceivably should deliver considerably higher readings. The sensitivity of a single urine POC-CCA was in the same order as that of a single Kato-Katz thick smear examination, while the sensitivity approached that of triplicate Kato-Katz when a combination of both CAA and CCA assays was used. The promising results from the current proof-of-concept study call for larger investigations that will determine the accuracy of the urine-based CCA and CAA assays for S. mekongi and S. japonicum diagnosis.

  18. 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.

  19. 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

  20. 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

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    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.

  1. 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.

  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. Extraction and Stability of Pigment from Hypericum japonicum%田基黄色素的提取及其稳定性

    Institute of Scientific and Technical Information of China (English)

    王海涛; 彭金龙; 张斌; 张璐

    2012-01-01

    Objective-. To investigate extraction methods of pigment from Hypericum japonicum by different solvents, and investigate effects of different factors on stability of pigment. Method; Pigment was extracted with 6 different solvents, stability of pigment was tested with pH, temperature, light, reducing agents and oxidants. Result: Extraction effect of pigment was optimum by ultrasonic extraction 10 min with 95% ethanol at 30 ℃ , These pigment was unstable in light, strong acidity and alkaline, metal ions of Mg2+ , Al3* and other conditions. Food additives, reducing agents and oxidants had little effect on stability of pigment. Conclusion; These, pigment had good stability, but it must prevent pigment using in light and avoid acidity and alkaline condition. Pigment from H. japonicum could be used as a good resource of natural plant pigment.%目的:探讨不同溶剂提取田基黄色素的方法,并考察各种因素对色素稳定性的影响.方法:采用6种不同溶剂提取色素,测试pH、温度、光熙、还原剂及氧化剂等因素对色素稳定性的影响.结果:田基黄色素用95%乙醇30℃超声提取10 min效果最佳,该色素在光照、强酸碱性、Mg2+、Al3+等条件下不稳定,食品添加剂、还原剂、氧化剂对色素的稳定性影响不大.结论:该色素具有较好稳定性,但须防止光照,避免酸碱性条件下使用.田基黄色素可作为一种较好的天然植物色素资源.

  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. MORPHOMETRICS OF TWO DIMENSIONS ON SCHISTOSOMA JAPONICUM EGG%日本血吸虫虫卵的二维数据测量

    Institute of Scientific and Technical Information of China (English)

    彭善友; 黎辉; 等

    2001-01-01

    目的 设计人门静脉内日本血吸虫虫卵过滤器孔眼的大小。 方法 采用图像分析仪测量虫卵的周长、面积、长径、短径、等效直径、圆球度。依据长径、短径数据算出虫卵的体积和均值。 结果 长径最大值84.55 μm,最小值39.19 μm;短径最大值57.22 μm,最小值23.2 μm。体积最大值131 039 μm3,最小值14 423 μm3。 结论 长径99%范围内的最小值45.49 μm,短径99%范围内的最小值27.43 μm,体积99%范围内的最小值17 817.59 μm3。本研究结果为设计人门静脉内日本血吸虫虫卵过滤器孔眼的大小提供了科学依据。%Objective To design the hole size of a filter for Schistosoma japonicum egg in human portal vein, measuring relative data of the eggs is needed. Methods The eggs were gotten form the rabbits infected with schistosomiasis japonicum. The perimeter, area, long diameter, short diameter, eguivalent diameter and sphere grade of each egg were measured by image analyser, then the volume of eggs and its average were worked out according to the long diameter and short diameter. Results Long diameter maximum is 84.55 μm, its minimum is 39.19 μm, short diameter max is 57.22 μm, its minimum is 23.2 μm, volume maximum is 131 039 μm3 and the minimum is 14 423 μm3. Conclusion Minimum in 99% range of the long diameter is 45.49 μm, minimum is 99% range of the short diameter is 27.43 μm, minimum in 99% range of the volume is 17 817.59 μm3. The results supplied scientific warranty for designing the hole size of the filter.

  7. 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

  8. Abundance and diversity of soybean-nodulating rhizobia in black soil are impacted by land use and crop management.

    Science.gov (United States)

    Yan, Jun; Han, Xiao Zeng; Ji, Zhao Jun; Li, Yan; Wang, En Tao; Xie, Zhi Hong; Chen, Wen Feng

    2014-09-01

    To investigate the effects of land use and crop management on soybean rhizobial communities, 280 nodule isolates were trapped from 7 fields with different land use and culture histories. Besides the known Bradyrhizobium japonicum, three novel genospecies were isolated from these fields. Grassland (GL) maintained a higher diversity of soybean bradyrhizobia than the other cultivation systems. Two genospecies (Bradyrhizobium spp. I and III) were distributed widely in all treatments, while Bradyrhizobium sp. II was found only in GL treatment. Cultivation with soybeans increased the rhizobial abundance and diversity, except for the soybean monoculture (S-S) treatment. In monoculture systems, soybeans favored Bradyrhizobium sp. I, while maize and wheat favored Bradyrhizobium sp. III. Fertilization decreased the rhizobial diversity indexes but did not change the species composition. The organic carbon (OC) and available phosphorus (AP) contents and pH were the main soil parameters positively correlated with the distribution of Bradyrhizobium spp. I and II and Bradyrhizobium japonicum and negatively correlated with Bradyrhizobium sp. III. These results revealed that different land uses and crop management could not only alter the diversity and abundance of soybean rhizobia, but also change interactions between rhizobia and legume or nonlegume plants, which offered novel information about the biogeography of rhizobia.

  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. Anti-obesity effect of Crinum asiaticum var. japonicum Baker extract in high-fat diet-induced and monogenic obese mice.

    Science.gov (United States)

    Jeong, Yong Joon; Sohn, Eun-Hwa; Jung, Yong-Hwan; Yoon, Weon-Jong; Cho, Young Mi; Kim, Inhye; Lee, Sung Ryul; Kang, Se Chan

    2016-08-01

    This study determined the anti-obesity effect of Crinum asiaticum var. japonicum Baker extract (CAE) on adipocytes and obese mice. The inhibitory effects of CAE on adipocyte differentiation and adipogenesis were determined using differentiation induction medium in 3T3-L1 cells. To get an insight into underlying molecular actions of CAE, we investigated the changes in the expression levels of genes involved in lipogenesis by CAE treatment using qRT-PCR. CAE strongly suppressed adipocyte differentiation through downregulation of PPARγ, C/EBPα, C/EBP β, and aP2. CAE treatment could also suppress the expression levels of ACC, FAS, LPL and HMGCR gene in 3T3-L1 cells. Male C57BL/6 strain and C57BL/6J-ob/ob strain mice were fed with HFD containing 60% fat and normal diet in the presence or absence of 25, 50, and 100mg/kg CAE for 7 weeks. CAE supplementation could highly suppress the body weight gain and epididymal fat accumulation without changes in food uptake in both obese models. Increases in total cholesterol, LDL-cholesterol and triglyceride were highly suppressed in the presence of CAE. In summary, CAE has an anti-obesity effect and this anti-obesity potential might be associated with downregulation of genes involved in adipocyte differentiation and lipogenesis.

  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. Effect of mefloquine administered orally at single, multiple, or combined with artemether, artesunate, or praziquantel in treatment of mice infected with Schistosoma japonicum.

    Science.gov (United States)

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

    2011-02-01

    The purpose of the study is to explore the efficacy of mefloquine administered orally at single, multiple doses, or in combination with artesuante, artemether, or praziquantel in mouse--Schistosoma japonicum model. A total of 205 mice were divided into 4 batches and each batch of mice was infected percutaneously with 40 S. japonicum cercariae for 35 days. The infected mice were treated orally with mefloquine at single doses, multiple daily doses, or combined with artesunate, artemether, or praziquantel, while infected but untreated mice served as control. All treated animals were killed 4 weeks post-treatment for assessment of effect. When infected mice were treated orally with mefloquine at single or multiple daily doses under the same total dose levels, the tendency to decrease the efficacy was seen. Particularly, when a lower single dose of 100 mg/kg was divided equally into five daily doses of 20 mg/kg, the efficacy decreased statistically significant (Pmefloquine or artesuante at a single dose of 100 mg/kg, a moderate effect against schistosomes was observed. No further significant reduction of total and female worm burdens was seen, when the two drugs combined together at the same dose level. On the other hand, administration of mefloquine combined with artesunate at single dose of 50 mg/kg, which exhibited no effect against schistosomes, resulted in significant reduction of total and female worm burdens in comparison with the groups treated with mefloquine and artesunate alone at the same dose level. Similar results were observed in treatment of infected mice with mefloquine in combination with artemether at the smaller dose of 50 mg/kg. The total worm burden was significantly lower than that of control and the female worm burden was also significant lower than that of groups treated with mefloquine and artemether alone. Interestingly, in administration of mefloquine 100 mg/kg combined with artemether 100 mg/kg to the infected mice, all female worms were

  13. 日本血吸虫非编码小RNA研究进展%Advances in small noncoding RNAs of schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    阳青兰(综述); 肖建华(审校)

    2016-01-01

    Small noncoding RNAs,a growing family of regulatory RNAs that includes micro-RNAs,endogenous small inter-fering RNA,Piwi-interacting RNA and competitive endogenous RNA,have recently emerged as cruicial components in many biological processes. Recent findings have provided the small noncoding RNAs expression to undergo dramatic changes in the schistosomiasis. Accumulated evidence suggests that host miRNAs may be involved in regulating the pathogenesis of schistosomiasis (miR-454 and miR-203). MiR-223 and miR-451 may be involved in regulating the development of schisto-some. Besides,recent work implies that schistosome-derived miRNAs may play critical regulatory roles in schistosome de-velopment and sexual maturation,egg production,as well as the pathogenesis of schistosomiasis (sja-let-7,sja-bantam). Moreover,sja-miR-3479 and sja-miR-0001 could be used as biomarkers for schistosomiasis diagnosis. Here,we summarize recent progress in regarding small noncoding RNAs in schistosoma japonicum. This may be helpful in search for potential new drugs,and for biomarkers in control of schistosoma japonicum infection leading to schistosomiasis in the future.%非编码小RNA是一类通过调节基因的表达从而影响生命活动的小分子,主要包括微小RNA、内源性小干扰RNA、Piwi蛋白相互作用的RNA及竞争性内源RNA。研究发现日本血吸虫感染动物后,宿主和虫体的非编码小RNA的表达水平均发生了改变。宿主源的非编码小RNA主要为微小RNA,其中miR-454和miR-203与日本血吸虫病发生发展有关,miR-223和miR-451与日本血吸虫自身生长发育有关;日本血吸虫非编码小 RNA包括微小RNA、内源性小干扰RNA,这些非编码小RNA在日本血吸虫的生长、发育、性成熟及产卵中发挥着重要作用,如sja-let-7、sja-bantam,而且像sja-miR-3479和sja-miR-0001还可以诊断日本血吸虫病。就日本血吸虫非编码小RNA的研究进展进行综述,为日本血吸虫病的防治提供参考。

  14. 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.

  15. 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

  16. 室内繁殖东方田鼠感染日本血吸虫的实验研究%Study of closed colony of Microtus fortis infected with Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    成钢; 曾文虎; 王京仁; 王文彬; 李淑红

    2013-01-01

    目的 比较室内繁殖东方田鼠与昆明鼠感染日本血吸虫后不同阶段脏器病变以及体内日本血吸虫发育与存活状况.方法 采用日本血吸虫尾蚴感染健康东方田鼠和昆明鼠,在感染后12、20、40d剖杀,观察并比较东方田鼠以及昆明鼠肝、肾、肺等脏器组织病变及体内日本血吸虫发育与存活情况.结果 日本血吸虫感染后12、20 d昆明小鼠和感染40d后东方田鼠的内脏器官均未出现明显病变;感染12 d及20d后的东方田鼠肝脏、肾脏及脾脏出现明显白色结节,且以感染12d的组织病变更为明显,部分东方田鼠病变组织仅见于肝脏.病理切片显示,病变肝脏和肾脏组织中存在完整日本血吸虫虫体,与正常组织界限分明;而无病变的肝脏和肾脏组织病理切片未发现虫体.结论 室内繁殖东方田鼠感染日本血吸虫后12d免疫反应剧烈且存在明显的个体差异.%Objective To understand the pathological changes of tissues and organs in Microtusfortis infected with Schistosoma japonicum for further insight into the biological dynamic variation and explore immune mechanisms of M.fortis against S.japonicum infection.Methods The healthy M.fortis and mouse were infected with cercariae of S.japonicum through abdominal skin.On the day 12,20 and 40,the M.fortis and mouse were sacrificed by cervical dislocation and dissected.The liver and kidney slices of M.fortis and mouse of the different groups were prepared and stained with hematoxylin eosin stain (HE) by conventional techniques,respectively.Results We did not find obvious lesions on the internal organs of mouse 12 d and 20 d after the infection and the lesions of M.fortis 40 d after the infection.However,white nodules appeared on the liver,kidney and spleen of the M.fortis 12 d and 20 d after the infection,and there were a lot of S.japonicum worms on pathological sections of inflamed liver and kidney tissues,and the boundary between worms and

  17. 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.

  18. 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

  19. 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.

  20. Bioinformatic analysis, cloning and expression of Schistosoma japonicum SCP/TAPS gene family%日本血吸虫SCP/TAPS基因家族的生物信息学分析和克隆表达

    Institute of Scientific and Technical Information of China (English)

    陈静芳; 胡旭初; 王乐旬; 余新炳

    2011-01-01

    目的 以黄蜂Vv Vesv5蛋白为检索源检索日本血吸虫EST数据库,对检索出的序列进行生物信息学分析,并挑选出有代表性的基因进行克隆表达,为日本血吸虫SCP/TAPS家族蛋白的功能研究打下一定的基础.方法 利用生物信息学软件对检索出的日本血吸虫SCP/TAPS家族蛋白进行生物信息学分析,经PCR确定后,以AAW24579为代表进行进一步的研究,将其命名为SjVAL2.根据SjVAL2的序列设计引物,通过RT-PCR扩增出全长编码区域,克隆到原核表达载体pET-28a(+)中,在大肠埃希菌中诱导表达并纯化,免疫印迹实验(Western-blotting)鉴定纯化的重组蛋白.结果 日本血吸虫SCP/TAPS家族中至少有17个不同的蛋白成员存在,多序列比对分析发现它们初步可以分为3个不同的亚组.以日本血吸虫成虫cDNA为模板RT-PCR扩增出SjVAL2序列,PCR、双酶切及DNA测序均证实pET-28a(+)-SjVAL2重组质粒构建成功.SDS-PAGE结果表明,目的基因在大肠埃希菌中获得高效表达,重组蛋白能识别日本血吸虫感染的小鼠血清,说明其具有免疫反应性.结论 日本血吸虫SCP/TAPS家族中至少有17个不同的蛋白成员存在,其中SjVAL2基因可在原核表达系统中高效表达,为进一步研究该家族蛋白的功能打下基础.%The Vespula vulgaris Wasp Venom Allergen 5 (Vv Vesv5, GenBank accession No. AAA30333) protein sequence was used in a tBLASTn search of S. Japonicum ESTs. Detail bioinformatic analysis were conducted and an interesting gene were cloned and expressed to investigate the role in the life cycle of S. Japonicum. After PCR confirmation of the sequences from parasite-derived cDNA, AAW24579 was selected for further study and named S. Japonicum Venom allergen-like transcript (SjVAL2). The coding region of SjVAL2 was amplified by RT-PCR using specific primers and cloned into prokary-otic expression vector pET-28a( +), then expressed in E. Coli BL2KDE3). The purified recombinant

  1. 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

  2. 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.

  3. 家鸭具有作为血吸虫转运宿主的可能性%STUDY ON THE POSSIBILITY OF DUCK AS TRANSPORT HOSTS FOR SCHIS TOSOMA JAPONICUM

    Institute of Scientific and Technical Information of China (English)

    沈立成; 杨艺; 陆珂; 李浩; 石耀军; 朱传刚; 王海生

    2012-01-01

    为研究日本血吸虫(Shistosomia japonicum)卵通过家鸭消化系统后的孵化能力,将人工感染血吸虫尾蚴45d的小鼠剖杀,取肝脏和小肠制成匀浆,并进行虫卵计数,将带有虫卵的肝脏及小肠匀浆液分别灌喂家鸭,观测不同时间收集鸭粪的毛蚴孵出情况。结果显示,经过家鸭体内2h后,粪便中孵化率达到峰值,相对孵化率分别为0.61%和4.066%;7h后,没有虫卵能孵化出毛蚴。同一批次的虫卵所孵化的毛蚴中,大约经过4h后其孵化率达到峰值。结果表明,日本血吸虫虫卵通过家鸭消化系统后仍有部分依然保持孵化能力,家鸭具有可能作为血吸虫的转运宿主,拥有传播血吸虫有效虫卵的能力。%To study the hatchability of Schistosomia japonicum eggs in duck digestive system, mice were infected with S.japonicum cercarias and sacrificed 45 days later. The liver or intestine homogenates containing eggs were fed to ducks. Duck faeces were collected to examine the hatchability of eggs using miracidia hatching technique. Viable eggs were recovered from all ducks although the hatchability significantly declined after the passage through ducks. The hatching peak reached at 2 hours post ingestion. The relative hatchability rates were 0.61% and 4.066% for liver or intestine homogenates. No miracidia were found at 7 hours after ingestion. The hatch rate reached its peak after approximately 4 hours' hatching although some miracidia were found at 12 hours. The eggs recovered from duck faeces maintained hatchability, which suggested that ducks might play a role in transmission of S.japonicum.

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. Chemical constituents ofAspergillus versicolor EN-298, an endophytic fungusderived from the marine algaCeramium japonicum%日本仙菜来源内生真菌Aspergillus versicolor EN-298化学成分研究

    Institute of Scientific and Technical Information of China (English)

    王佳宁; 李晓明; 张鹏; 徐刚明; 王斌贵

    2015-01-01

    从采自烟台养马岛的日本仙菜(Ceramium japonicum)中分离得到的一株内生真菌Aspergillus versicolor EN-298,对其次级代谢产物进行了系统研究,利用反相柱层析、正相柱层析、葡聚糖凝胶Sephadex LH-20以及制备薄层层析(pTLC)等分离手段获得10个单体化合物。通过一维、二维核磁共振技术(1D、2D NMR)等鉴定了所有化合物的结构。并对分离得到的化合物进行了抑菌活性测试,其中化合物10对迟缓爱德华氏菌(Edwardsiella tarda)表现出一定的抑制作用。%Ten compounds were isolated from the secondary metabolites ofAspergillus versicolor EN-298, an endophytic fungus that was isolated from the marine red algaCeramium japonicum. The structures of the isolated compounds were elucidated by means of spectroscopic methods including UV and NMR (1D and 2D) as well as by comparison with authentic standards and literature reports. All compounds were evaluated for antimicrobial activity, and the compound10 showed moderate activity against aqua-bacterialEdwardsiella tarda with an MIC value of 16μg/mL.

  10. 白头翁总皂苷对日本血吸虫虫卵毛蚴尾蚴的作用%Effect of Pulsatilla chinensis (Bunge) Regel saponins on eggs, miracidia and cercariae of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    陈岩勤; 许琼明; 诸葛洪祥; 梁幼生; 李笑然; 杨世林

    2013-01-01

    目的 观察白头翁总皂苷(Pulsatilla chinensis(Bunge)Regel saponins,PRS)对日本血吸虫虫卵、毛蚴、尾蚴的杀灭效果,为白头翁总皂苷作为抗血吸虫新药提供理论和实验依据.方法 采用尾蚴腹部贴片法感染ICR小鼠,感染后42 d的鼠肝脏经研磨、过筛获得虫卵,毛蚴由虫卵孵化,尾蚴由阳性感染钉螺光照逸出,采用0、1、2、3、4、5、6、7、8 μg/ml的PRS药液分别于不同时间作用虫卵、毛蚴、尾蚴.结果 不同浓度PRS及对照药物吡喹酮(PZQ)对日本血吸虫虫卵作用24 h的孵化结果显示,PRS对虫卵孵化的抑制效果略优于PZQ,尤其在4μg/ml浓度时,日本血吸虫虫卵对PRS的作用更敏感.1、2、3、4、5、6、7、8 μg/ml的PRS药液作用于毛蚴30 min后,毛蚴死亡率分别为13.47%、26.05%、60.99%、90.84%、100%、100%、100%、100%.作用于尾蚴30 min后,尾蚴的死亡率分别为5.32%、18.81%、44.7%、76.87%、98.28%、100%、100%、100%.毛蚴、尾蚴的死亡率对PRS的作用时间和浓度有一定依赖性.结论 体外实验显示PRS对日本血吸虫虫卵、毛蚴、尾蚴均有杀伤作用,有望成为新的抗血吸虫药物.%Objective To investigate the effect of Pulsatilla chinensis (Bunge) Regel saponins (PRS) against the eggs, mira-cidia, cercariae of Schistosoma japonicum in vitro and compare its efficacy with praziquantel. Methods ICR mice were infected with the cercariae of S. japonicum by the patching abdominal method. The livers of the mice were grinded, screened, and then the eggs of S. japonicum were obtained 42 days post-infection. The miracidia were hatched by using the eggs, and the cercariae were obtained by using the infected Oncomelania snails on the light. The eggs, miracidia and cercariae of S. japonicum were incubated with 0,1,2,3,4,5,6,7,8 μg/ml PRS for different time, and praziquantel (PZQ) was used as the control. Results PRS suppressed the hatching rates of eggs for

  11. Significance of higher drug concentration in erythrocytes of mice infected with Schistosoma japonicum and treated orally with mefloquine at single doses.

    Science.gov (United States)

    Tao, Yi; Xue, Jian; Jiang, Bin; Zhang, Hao-Bing; Xiao, Shu-Hua

    2015-12-01

    The purpose of the present study is to understand the pharmacokinetic feature of mefloquine measured by erythrocytes and plasma in Schistosoma japonicum (S. j.)-infected mice and non-infected mice after oral administration of the drug at single doses. A high-performance liquid chromatography (HPLC) method was used to measure the plasma and erythrocyte concentrations of mefloquine at varying intervals posttreatment. Our results demonstrated that in non-infected mice treated orally with mefloquine at an ineffective dose of 50 mg/kg or effective dose of 200 mg/kg for 2-72 h, the erythrocyte-to-plasma ratios of mefloquine were 5.8-11.2 or 2-14.2. On the other hand, in S. j.-infected mice treated with the same single doses of the drug, the erythrocyte and plasma drug concentration ratios were 3.1-4.6 or 2.9-8.5, manifesting that either in infected mice or in non-infected mice that received oral mefloquine resulted in higher concentration of mefloquine in erythrocytes than that in plasma. Unexpectedly, under oral administration of mefloquine at a higher single dose of 200 mg/kg, the pharmacokinetic parameter C max values for plasma from S. j.-infected and non-infected mice were 1.6 ± 0.3 and 2.0 ± 0.4 μg/mL, respectively, which were below the determined in vitro LC50 (50 % lethal concentration) value of 4.93 μg/mL. Therefore, the plasma concentration of mefloquine may display a little effect against schistosomes during the treatment. Although the values of T 1/2 and AUC0-∞ for erythrocytes were significantly longer and higher in infected mice than those of corresponding non-infect mice that received the same single mefloqine dose of 50 mg/kg, the C max value was only 2.6 ± 0.4 μg/mL lower than the determined in vitro LC50, which may explain why this low single dose is ineffective against schistosomes in vivo. After administration of higher mefloquine dose of 200 mg/kg, the C max value for erythrocytes in infected mice was 30 % (7.4 ± 0

  12. 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

  13. In vitro effects of EGCG on Schistosoma japonicum cercariae%表没食子儿茶素没食子酸酯(EGCG)体外抗日本血吸虫尾蚴作用的研究

    Institute of Scientific and Technical Information of China (English)

    冯金梅; 袁发浒; 刘锴; 张泽华; 胡松; 邱文洪; Stephen Hsu; 宋文剑

    2016-01-01

    日本血吸虫尾蚴具有体外杀伤作用,发生的形态结构可能影响其侵袭宿主皮肤的能力.%Objectives To investigate whether epigallocatechin-3-gallate (EGCG) kills Schistosoma japonicum cercariae in vitro and its effects on the ultrastructure of those cercariae in order to evaluate whether EGCG protects against infection with S.japonicum cercariae.Methods Cercariae of S.japonicum were placed in distilled water (control group) or dif ferent concentrations of EGCG (0.2% and 1 %).Cercariae were treated for 30 and 60 min and then observed with a dissecting microscope.Cercariae that were treated for 30 min were also observed using scanning electron microscopy.Male BALB/c mice,6-8 weeks of age,were randomly divided into tlree groups with six animals each:group Ⅰ (control group,infected with S.japonicurn cercariae treated with 0% EGCG for 30 min),group Ⅱ (infected with S.japonicum cercariae treated with 0.1 % EGCG for 30 min),and group Ⅲ (infected with S.japonicum cercariae treated with 1% EGCG for 30 min).All experimental mice were anesthetized and percutaneously infected with 30 cercariae of S.japonicum that were placed on shaved abdominal skin.The animals were maintained in conventional animal housing at the Labo ratory Animal Center of Jianghan University,China.All mice were given standard rodent food pellets and water ad libitum.At six weeks post-infection,the mice were sacrificed and adult worms of S.japonicum were collected,and the rate of infection,the average worm burden,and the rate of worm clearance were calculated.Results Under a dissecting microscope,S.japonicum cercariae treated with 0% EGCG for 30 min (the controls) had a normal morphology and they moved freely,while cercariae treated with 0% EGCG for 60 min were rigid,curled up into a ball,and not moving.Cercariae that were treated with 0.2 % or 1% EGCG for 30 min were highly active,their tails lashed rapidly,they twitched,and their bodies were deformed

  14. Genetic diversity of Rhizobia isolates from Amazon soils using cowpea (Vigna unguiculata) as trap plant.

    Science.gov (United States)

    Silva, F V; Simões-Araújo, J L; Silva Júnior, J P; Xavier, G R; Rumjanek, N G

    2012-04-01

    The aim of this work was to characterize rhizobia isolated from the root nodules of cowpea (Vigna unguiculata) plants cultivated in Amazon soils samples by means of ARDRA (Amplified rDNA Restriction Analysis) and sequencing analysis, to know their phylogenetic relationships. The 16S rRNA gene of rhizobia was amplified by PCR (polymerase chain reaction) using universal primers Y1 and Y3. The amplification products were analyzed by the restriction enzymes HinfI, MspI and DdeI and also sequenced with Y1, Y3 and six intermediate primers. The clustering analysis based on ARDRA profiles separated the Amazon isolates in three subgroups, which formed a group apart from the reference isolates of Bradyrhizobium japonicum and Bradyrhizobium elkanii. The clustering analysis of 16S rRNA gene sequences showed that the fast-growing isolates had similarity with Enterobacter, Rhizobium, Klebsiella and Bradyrhizobium and all the slow-growing clustered close to Bradyrhizobium.

  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. Risk factors of Schistosoma japonicum infection in Xingzi County%星子县居民血吸虫感染危险因素分析

    Institute of Scientific and Technical Information of China (English)

    肖国良; 翟敏玲; 陶波; 姜秋林; 刘建平; 习青华; 周宪民; 卢曲琴

    2016-01-01

    目的:分析江西省星子县居民血吸虫感染的危险因素。方法2013年采用整群抽样的方法从江西省星子县血吸虫病流行区随机抽取6个行政村进行调查,并采用Kato⁃Katz法对这6个村≥5岁的全部常住居民进行病原学检查;应用单因素χ2检验和多因素Logistic回归模型对影响血吸虫病流行的相关危险因素进行分析。结果2013年共粪检调查2050人,查出粪检阳性146人,血吸虫感染率为7.1%。单因素χ2检验显示,不同性别、年龄、文化程度、职业人群血吸虫感染率差异均有统计学意义(χ2=26.485、16.836、25.700、90.805,P均<0.05)。多因素Logistic回归分析显示,男性感染血吸虫的风险是女性的3.041倍;小学及以下文化程度人群血吸虫感染风险是大专及以上文化程度人群的8.870倍;初中和高中文化程度人群感染血吸虫的风险分别是大专及以上文化程度人群的5.598倍和2.995倍;渔民血吸虫感染的可能性最高,是其他职业的3.053倍。结论影响星子县居民血吸虫感染的危险因素主要包括性别、职业和文化程度;应强化血吸虫病健康教育,加强疫水接触防护工作,减少血吸虫感染发生。%Objective To explore the risk factors of Schistosoma japonicum infection in the residents in Xingzi County,Ji⁃angxi Province. Methods Six administrative villages from different areas were randomly selected by the cluster sampling meth⁃od as the study sites in Xingzi Country in 2013,and all the residents aged 5 years or above were investigated epidemiologically, and the schistosome infection was surveyed by Kato⁃Katz technique. The risk factors of schistosomiasis were analyzed by using the Chi⁃square test analysis and multivariate Logistic regression model. Results In 2013,there were 2 050 residents received the stool examination and 146 persons were positive,the schistosome infection rate was 7.1%. The Chi

  17. 日本血吸虫成虫吡喹酮用药前后差异表达序列的筛选%Screening of differential expression of expressed sequence tags (ESTs) in adult Schistosoma japonicum treated with praziquantel

    Institute of Scientific and Technical Information of China (English)

    周水茂; 江元山; 陈智; 熊燕; 田俊华; 周敦金; 徐明星; 孙东光

    2011-01-01

    Objective To establish subtracted cDNA libraries using the suppression subtractive hybridization (SSH)technique based on the livers of New Zealand white rabbits infected with Schistosoma japonicum before and after praziquantel treatment, providing the basis for screening the drug-response molecular target during the therapy.Methods New Zealand white rabbits infected with S.japonicum were assigned to two groups to receive praziquantel medication or blank, respectively.The SSH technique was conducted to build the forward and reverse subtracted cDNA libraries using the PCR-Select cDNA Subtraction kit.The differentially expressed ESTs were screened and positive clones were isolated from the libraries for bioinformatic analysis.Results Thirty-nine positive clones were isolated from two subtracted cDNA libraries, including 22 from the forward library and 17 from the reverse library.The analysis revealed that the ESTs were mainly responsible for coding enzymes and proteins associated with protein synthesis and degradation.Conclusion The subtracted cDNA libraries before and after praziquantel treatment for S.japonicum were successfully constructed, laying a solid foundation for further research into the molecular mechanisms of this medication therapy.%目的 应用抑制性消减杂交(SSH)技术构建吡喹酮治疗日本血吸虫感染新西兰大白兔前后肝脏内差减cDNA文库,为筛选吡喹酮治疗过程中日本血吸虫体内药物反应分子靶标奠定基础.方法 日本血吸虫感染新西兰大白兔,吡喹酮治疗者为实验组,未经吡喹酮治疗者为对照组,应用PCR-Select cDNA Subtraction试剂盒进行SSH分析,分别构建正向和反向消减cDNA文库,对文库进行筛选,挑取阳性克隆测序获得差异表达EST或新EST,并进行生物信息学分析.结果 从2个消减文库中共筛选到39个有效阳性克隆,其中正向文库22个,反向文库17个,分析表明这些EST编码主要是一些酶及与蛋白质合成和降

  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. Research progress on transmission capacity of reservoir host of Schistosoma japonicum%日本血吸虫保虫宿主传播能量研究进展

    Institute of Scientific and Technical Information of China (English)

    汪奇志; 汪天平; 张世清

    2013-01-01

    本文对牛、羊、猪、犬、猫、野鼠等日本血吸虫保虫宿主传播能量的相关研究进行了综述,并结合当前实施的以传染源控制为主的综合防治策略,提出了在人和牛作为传染源的作用逐渐减弱的情况下,对非牛家畜(羊、犬、猪等)和野鼠等动物宿主的实际传播能量进行重新评价,以及探索血吸虫传播环节中相关传播阈值等研究方向.%This paper reviews the researches related to the reservoir hosts of Schistosoma japonicum, including bovine, sheep, dog, cat and wild mouse. Combined with the implementation of the comprehensive control strategy with an emphasis on infection source control in current, it puts forward several future research directions under the condition that the effects of people and bovine as infection sources weakened gradually, such as revaluation on the actual transmission capacity of non-bovine livestock, for example, sheep, dog, pig and wild mouse, as well as discussion on some transmission threshold values in the infection link of Oncomelania snails.

  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. Diversity and biogeography of rhizobia isolated from root nodules of Glycine max grown in Hebei Province, China.

    Science.gov (United States)

    Li, Qin Qin; Wang, En Tao; Zhang, Yun Zeng; Zhang, Yan Ming; Tian, Chang Fu; Sui, Xin Hua; Chen, Wen Feng; Chen, Wen Xin

    2011-05-01

    A total of 215 rhizobial strains were isolated and analyzed with 16S rRNA gene, 16S-23S intergenic spacer, housekeeping genes atpD, recA, and glnII, and symbiotic genes nifH and nodC to understand the genetic diversity of soybean rhizobia in Hebei province, China. All the strains except one were symbiotic bacteria classified into nine genospecies in the genera of Bradyrhizobium and Sinorhizobium. Surveys on the distribution of these rhizobia in different regions showed that Bradyrhizobium japonicum and Bradyrhizobium elkanii strains were found only in neutral to slightly alkaline soils whereas Bradyrhizobium yuanmingense, Bradyrhizobium liaoningense-related strains and strains of five Sinorhizobium genospecies were found in alkaline-saline soils. Correspondence and canonical correspondence analyses on the relationship of rhizobial distribution and their soil characteristics reveal that high soil pH, electrical conductivity, and potassium content favor distribution of the B. yuanmingense and the five Sinorhizobium species but inhibit B. japonicum and B. elkanii. High contents of available phosphorus and organic matters benefit Sinorhizobium fredii and B. liaoningense-related strains and inhibit the others groups mentioned above. The symbiotic gene (nifH and nodC) lineages among B. elkanii, B. japonicum, B. yuanmingense, and Sinorhizobium spp. were observed in the strains, signifying that vertical gene transfer was the main mechanism to maintain these genes in the soybean rhizobia. However, lateral transfer of symbiotic genes commonly in Sinorhizobium spp. and rarely in Bradyrhizobium spp. was also detected. These results showed the genetic diversity, the biogeography, and the soil determinant factors of soybean rhizobia in Hebei province of China.

  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. Study on Structure and Function of Class Ⅰ Polypeptide Release Factor C Domain in Blepharisma japonicum%日本赭纤虫第一类肽链释放因子C端结构域分析

    Institute of Scientific and Technical Information of China (English)

    陈洁; 梁爱华; 杨斌盛

    2012-01-01

    eRF1的C结构域与eRF3的C结构域相互作用对于蛋白质翻译终止过程中的快速反应至关重要.通过计算机同源建模对日本赭纤虫第一类肽链释放因子C结构域Bj-eRF1C进行结构模拟,发现在Bj-eRF1C结构域中有些肽段直接参与了eRF1-eRF3相互作用,特别是Bj-eRF1C结构域中V294和D297位点高度保守.通过定点突变与pull-down分析,表明在Bj-eRF1C结构域中V294和D297是eRF1-eRF3相互作用的关键位点.%The C domain of eRFl interacts with the C domain of eRF3. This interaction is essential for fast kinetics of the protein translation termination. In this study, the eRFl-eRF3 interaction binding sites on class I polypeptide release factor C domain in Blepharisma japonicum (Bj-eRFIC ) were stimulated based on homology modeling. Point mutation analysis were carried out. The results showed that V294 and D297 in Bj-eRFIC domain played an important role on eRFl-eRF3 interaction.

  7. 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.

  8. 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

  9. Preliminary investigation on in vitro-cultivation and manipulation of Schistosoma japonicum sporocysts%日本血吸虫胞蚴的体外培养和操作初探

    Institute of Scientific and Technical Information of China (English)

    江艳; 龚仁敏; 杨艺; 李小红; 刘毅; 朱传刚; 陆珂; 郑浩

    2011-01-01

    目的 探究一种体外培养日本血吸虫胞蚴的新方法,并通过体外注射,感染阴性钉螺,培养阳性钉螺.方法 将日本血吸虫虫卵孵化的毛蚴与作为滋养层的草地夜蛾(Spodoptera frugiperda)卵巢细胞(sf9细胞)共培养,观察毛蚴的生长情况,并拍摄生长照片.用显微注射针将3~5只母胞蚴注射到钉螺体内,计算钉螺的存活率及尾蚴的阳性率,用钉螺逸出的尾蚴人工感染昆明鼠,检测尾蚴的活力,用组织化学染色法和PCR方法鉴定尾蚴的相关基因特性.结果 毛蚴培养至第12小时观察到纤毛板已脱落,培养2~3 d的虫体呈不断的伸缩运动,此后母胞蚴维持该状态并不转化成子胞蚴.3次实验的钉螺存活率依次为24.72%、28.23%和57.89%,活螺阳性率依次为4.55%、8.57%和14.29%.阳性钉螺释放的尾蚴具有感染性,且具有与日本血吸虫相同的基因特性.结论 体外培养毛蚴通过显微注射可以人工感染钉螺并逸出有感染活力的尾蚴,为日本血吸虫幼虫的体外操作提供一种新的方法.%Objective To explore a new method of in vitro cultivation of Schistosoma japonicum sporocysts,infection of negative snails through in vitro-injection,and cultivation of positive snails.Methods Isolated miracidia were directly cultivated in sf9 cells-conditioned medium,the growth of miracidia was observed and the growth picture was taken.3-5 mother sporocysts were inoculated into the snails body using a microscopic needle.The survival rate of the snails and the positive rate of cercariae were calculated.Kunming mice were infected with the cercariac and the activity of cercariae Was detected.The related gene characteristics of cercariae was identified by histochemical examination and PCR method.Results The pictures of miracidia were recorded after co-cultivation for 1 h,12 h,24 h and 3 d.The shedding of the cilia of miracidia occurred within 12 h,parasites exhibited continuous stretching movements after

  10. Effect of a suspension of niclosamide on cercariae of Schistosoma japonicum and prevention of infections%氯硝柳胺悬浮剂灭蚴和预防血吸虫感染的研究

    Institute of Scientific and Technical Information of China (English)

    神学慧; 戴建荣; 李叶芳; 梁幼生; 李洪军; 唐建霞; 陈祥平

    2011-01-01

    Objective To investigate the effect of a suspension of niclosamide (SN) on cercariae of Schistosoma japonicum and effective concentrations of that suspension in order to provide scientific evidence for control strategies to prevent acute schistosomiasis. Methods SN was weighed and formulated into solutions with active concentrations of niclosamide of 1×10-5, 0. 5×10-5, 1×10-6, 0.5×10-6, 1× 10-1, and 0. 5×10-7 ml/100 ml in the lab, and the effect of SN on cercariae was determined. Solutions with an active concentration of niclosamide of 1. 2× 10-5, 1. 2× 10-6, 1. 2 ×10-7, 1.2×10-8, 1.2×10-0, and 1. 2×10-10 were formulated. Cercariae were counted 0, 10 and 30 min after 200 S. japonicum cercariae were transferred to the solutions and 30 min after mice were infected with the cercariae. Forty-five days after infection, the mice were dissected and the state of infection was observed. SN with a concentration of 0. 02 g/ m2 was sprayed for 1 h on the surface of water in the field, and then the infectivity of water was determined using sentinel mice. Results All cercariae were killed after coming into contact with an active concentration of niclosamide of more than 0. 5× 10-6 for 1 min, an active concentration of niclosamide of more than 0. 5× 10-7 for 2 min, or an active concentration of niclosamide of more than 1× 10-8 for 30 min. However, all survived after coming into contact with an active concentration of niclosamide of less than 0. 5 × 10-8 for under 60 min. After the cercariae came into contact with niclosamide of 1. 2 × 10-5 for 10 or 30 min, the cercariae failed to infect mice. When the cercariae were exposed to niclosamide of 1. 2 × 10-6 for 10 min, 80% of mice were infected, and the worm burden of mice was 3. 2 worms per mouse, but 30 min later the rate of infection in mice decreased to 0. Water in the field was infective, with a rate of infection in sentinel mice of 33. 33%. After 0. 02 g/m2 SN was sprayed on the water surface for 1 h, water

  11. 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

  12. 手性氟虫腈对意大利蜜蜂和稻螟赤眼蜂的急性毒性及安全评价%Acute Toxicities and Safety Evaluation of Chiral Fipronil to Apis mellifera L.and Trichogramma japonicum Ashmead

    Institute of Scientific and Technical Information of China (English)

    苍涛; 王新全; 王彦华; 吴长兴; 吴声敢; 陈丽萍; 俞瑞鲜; 赵学平

    2012-01-01

    手性农药多以外消旋体形式用于农业生产和卫生害虫防治中.但手性农药对映体通常具有不同的生物活性和毒性,这种现象在农药环境风险评价过程中往往被忽视.本研究以氟虫腈为例,首先用高效液相色谱-手性固定相(HPLC-CSP)技术拆分出氟虫腈的S型和R型2种对映体,分别采用点滴法和药膜法测定了氟虫腈对映体及外消旋体对意大利蜜蜂(A pis mellifera L.)和稻螟赤眼蜂(Trichogramma japonicum Ashmead)的急性毒性.结果显示,S(+)-氟虫腈、R(-)-氟虫腈和外消旋体对意大利蜜蜂的48 h-LD50分别为0.00341、0.00396和0.00383 μg·蜂-1,对稻螟赤眼蜂的24 h-LR50分别为7.56×10-7、8.06×10-7和7.29 ×10-7 mg·cm-2.研究表明,氟虫腈对意大利蜜蜂具有高毒性风险,对稻螟赤眼蜂有极高毒性风险,且氟虫腈对意大利蜜蜂和稻螟赤眼蜂的急性毒性无明显的对映体选择性.因此,使用氟虫腈单一对映体不会降低其对环境生物的毒害风险.%Most of the chiral pesticides were used as racemic forms in agricultural production and health pest control.However,differences existed in the biological activity and toxicity of the enantiomers of chiral pesticides,and the phenomena were usually ignored during the process of environmental risk evaluation of chiral pesticides.In this study,fipronil was selected as a model chiral pesticide,and its two enantiomers were isolated using an HPLC chiral stationary phase method (HPLC-CSP).The acute toxicities of S(+ )-fipronil,R(-)-fipronil and racemic fipronil to Apis mellifera L.and Trichogramma japonicum Ashmead were investigated by the standard drop method and drug membrane method.Results showed that the 48 h-LD50 of S( + )-fipronil,R( - )-fipronil and racemic fipronil to A .mellifera were 0.00341,0.00396 and 0.00383 μg· bee-1,respectively.The 24 h-LR50 of S( + )-fipronil,R( - )-fipronil and racemic fipronil to T.japonicum were 7.56 × 10-7,8.06× 10-7 and 7

  13. 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

  14. DNA polymorphism in the partial mitochondrial atp6 gene among Schistosoma japonicum isolates from Hunan Province%湖南省日本血吸虫线粒体atp6基因部分序列的多态性分析

    Institute of Scientific and Technical Information of China (English)

    夏英定; 汪世平; 李娟; 吴昌义; 田智; 尹铁球; 周云飞; 张树菊; 冯其梅

    2011-01-01

    目的 研究湖南省日本血吸虫不同地域自然隔离群线粒体ATP合成酶F0亚单位6基因(atp6)部分序列的遗传多态性,为湖南不同地域自然隔离群日本血吸虫的遗传特性研究提供实验依据.方法 采用试剂盒抽提基因组总DNA,以特异性引物对线粒体atp6基因进行PCR扩增,通过单链构象多态性技术(SSCP)筛选出差异带型并进行测序,用DNAStar 5.0及Mega 4.0软件进行比对分析.结果 湖南省5个流行区的日本血吸虫PCR扩增后获得了483 bp atp6部分序列,检测出17个变异位点,变异率为3.52%,雌雄虫之间的差异为0.0%~1.3%,不同地理来源虫株间的差异率为0.0%~1.5%.聚类分析结果表明:湖南不同地域自然隔离群日本血吸虫的种系发育不能按照其来源归为相应的系统发育进化树.结论 湖南不同地域自然隔离群日本血吸虫的atp6基因存在明显的个体差异,是否由于生存环境的选择压力造成了同一地理来源日本血吸虫个体之间atp6基因的遗传差异,其原因尚有待于进一步研究.%The aim of this study was to examinc sequence variation in the partial mitochondrial ATP synthase F0 subunit atp6 gene of Schistosoma japonicum and to provide foundation for studying the characterization and genetic structure of S. japonicum from different endemic origins of Hunan province. After extracting genomic DNA, atp6 was amplified with specific primers and screened by SSCP technique. Representative samples with variable banding patterns were selected for sequencing.DNAStar 5.0 and Mega 4.0 were used to analyze the sequences subsequently. Results showed that partial atp6 gene of S. japonicum was 483 bp, and 17 mutation sites were detected (3. 52%). Sequence variations for the partial atp6 were 0.0% -1.3% between male and female parasites, and 0.0%-1.5 % among isolates from different geographical origins. Phylogenetic analysis revealed that S. japonicum in Hunan province could not be

  15. 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

  16. Effect of bacterial root symbiosis and urea as source of nitrogen on performance of soybean plants grown hydroponically for bioregenerative life support systems (BLSSs)

    OpenAIRE

    Roberta eParadiso; Roberta eBuonomo; Dixon, Mike A.; Giancarlo eBarbieri; Stefania eDe Pascale

    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 r...

  17. 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.

  18. 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.

  19. 重组日本血吸虫磷酸丙糖转移酶免疫保护性的研究%Preliminary study of recombinant the triosephosphate transferase in immuno-protection of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    陈多玲; 张婧; 姚涌; 汪学龙

    2012-01-01

    Objective To observe the immuno-protection in mice attacked with cercaria for a new vaccine candidate of Schistosoma japonicum, the recombinant protein triosephosphate transferase (SjTPM). Methods The BALB/c mice were immunized with the recombinant SjTPM in the dose of 50μg/100μl、 100μg/100μl and 200μg/100μl, the saline solution was used for control group, then attacked with cercaria. The percentage of worm reduction and egg reduction was calculated to evaluate the immuno-protection effect. HE staining method was used to detect the hepatic changes and granulomatous changes in the liver tissue. Results The percentage of worm reduction was 27.1%,32.1% and 35.2%, respectively, in the experimental groups. The number of eggs in liver tissue was reduced by 34.2% ,39.29% ,43.66%, respectively. There were remarkable difference among the experimental groups and control group by t-test. There was no remarkable difference between the experimental groups by X2-test. Gross observation showed that the livers in control groups were dark brown and hard, with miliary nodules or worm eggs diffused in the livers. The recombinant SjTPM group has some extent of amelioration, the livers were bright red, relative soft and smooth, with less nodules of worm eggs in livers. Hepatic histological section demonstrated that there're generous large granulo- degeneration and necrosis of hepatic cells. The recombinant SjTPM vaccine group reversed the hepatic injury better than the control group. The structure of hepatic lobule was integrated on the whole, and the inflammatory reaction around the worm egg granulomas was light. Conclusion The molecular vaccine of rSjTPM had some immuno-protection effect.It suggested that rSjTPM was a potential candidate of S.japonicum.%目的 观察重组日本血吸虫磷酸丙糖转移酶(rSj-TPM)在小鼠抗血吸虫感染中的免疫保护作用,寻找新的血吸虫疫苗候选分子.方法 以rSj-TPM每鼠50μg/100μl、100μg/100μl和200μg/100μl

  20. 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

  1. 日本血吸虫26kDa谷胱甘肽S-转移酶基因真核表达载体的构建及序列测定%CONSTRUCTION AND SEQUENCING OF 26-KILODALTON GLUTATHIONE TRANSFERASES GENE OF SCHISTOSOMA JAPONICUM INTO EUKARYOTIC EXPRESSION VECTOR

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    目的扩增日本血吸虫26kDa谷胱甘肽S-转移酶(Sj26GST)基因片段,构建其重组真核表达载体,以研制SjGST核酸疫苗。方法根据已知基因序列设计一对引物,运用RT-PCR技术扩增Sj26GST目的基因片段,将其克隆到pcD-NA3质粒中,并经酶切、PCR鉴定,测序证实。结果获得GST-pcDNA3重组克隆。结论:本实验获得Sj26GST重组克隆,为进一步研制核酸疫苗创造了条件。%Aim To amplify the gene of 26-Kilodalton glutathione S-transferases of Schistosoma japonicum and construct a recombinant eukaryotic expression vector. Methods One pair of primers was designed according to the sequence of GST. The gene fragment of Sj26GST was obtained by RT-PCR,then it was cloned into a vector pcDNA3. It was corroborated through restriction enzymes cleaving,PCR and sequencing. Results GST-pcDNA3 recombinant clone was constructed successfully. Conclusions Sj26GST is a promising candidate of vaccine for Schistosoma japonicum. GSTrecombinant clone was achieved in the experiment. It is prepared for further study for DNA vaccine.

  2. 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.

  3. Extraction of IgG from serum of rabbit infected with Schistosoma japonicum by saturated ammonium sulphate%硫酸铵法提取血吸虫感染兔血清IgG初探

    Institute of Scientific and Technical Information of China (English)

    兰炜明; 姜唯声; 谢曙英; 徐银; 戴坤教; 高晓晖; 曾小军

    2016-01-01

    Objective To explore the feasibility of preparing the schistosomiasis positive reference product by the method of ammonium sulfate precipitation to extract the IgG from the infected rabbits ’ serum of Schistosoma japonicum. Methods We used the saturated ammonium sulfate solution to precipitate and separate the IgG from the infected rabbits ’ serum. Compared the titer of infected rabbits’ serum with the separated IgG,and compared unfreeze-drying titer of IgG with the freeze-drying by IHA,also studied the stability of freeze-drying IgG after we dissolve with water. Results The titer of the infected rabbits’ serum was 1:320 used to extract IgG by the method of ammonium sulfate precipitation,we dialysised,concentrated the liquid of the extracted IgG to the original volume,the extracted IgG titer was 1:320. the negative rabbits’ serum and sucrose as the protein protective agent added to the extracted IgG to freeze-drying,the extracted IgG titer of the unfreeze-drying and the freeze-drying were 1:160. The titer of the freeze-drying IgG dissolved in water changed with the time,the 0th,3rd,6th and 8th titer is 1:160,1:80,1:40 and 1:40 respectively. Conclusion All of the process of the ammonium sulfate precipitation method to extract the IgG and the freeze-dry-ing can not chang the titer of the antibody,but the process adding the protein protective agent to the extracted IgG reduced the titer,the titer of freeze-drying IgG dissolve with water falled after the 3rd. The ammonium sulfate precipitation method is suitable for extracting the IgG from the infected rabbit serum.%目的:探索硫酸铵法提取血吸虫感染兔血清IgG用于制备阳性参考品的可行性。方法采用饱和硫酸铵溶液对感染兔血清中IgG进行沉淀分离,IHA试剂测定分离前后、冻干前后的抗体滴度以及冻干IgG复溶后的稳定性。结果滴度为1:320的血吸虫感染兔血清经硫酸铵沉淀法提取IgG,经透析、浓缩至

  4. An investigation of polymorphism in the tetraspanin-2 gene of Schistosoma japonicum%日本血吸虫被膜蛋白Sj-TSP2基因序列多态性分析

    Institute of Scientific and Technical Information of China (English)

    张丽丽; 徐新东; 潘卫庆

    2011-01-01

    Objective To identify the polymorphic sites of Schistosoma japonicum tegument protein tetraspanin-2 (Sj-TSP2).Methods Paired worms were separated in cold PBS with the help of forceps under a microscope.Total RNA of individual worms was extracted by Trizol reagent.First strands cDNAs were synthesized using reverse transcriptase.The open reading frame(ORF) of Sj-TSP2 was amplified by PCR.The resulting PCR products were sequenced after inserted into the pBlu2KSM vector between BamH I and EcoR I sites.DnaSP was used to calculate the alleles and the polymorphic sites.Mega 4 was used to build Neighborjoining tree and calculate the ratio of non-synonymous to synonymous substitutions (dN/dS).Results 46 fulllength ORFs of Sj-TSP2 were successfully amplified by RT-PCR from 60 individual worms.In total,92 fulllength sequences were obtained from 46 worms,containing 20 unique alleles.20 alleles were generated by point mutation and Sj-TSP2-10 was the predominant allele.There were a total of 85 polymorphism sites.Of the 63 resulting codon changes,27 gave rise to amino acid substitution (non-synonymous changes).Among them,22 amino acid substitutions were located in the extracellular loop 2.Conclusion Sj-TSP2 was an extensive polymorphic gene with 20 alleles identified.%目的 分析日本血吸虫Sj-TSP2序列多态性.方法 抽提单条日本血吸虫成虫的总RNA,逆转录合成cDNA,PCR扩增目的片段.将目的片段克隆到载体pBlue2KSM中转化大肠杆菌,挑取10个阳性克隆测序.将获得的Sj-TSP2序列导入DnaSP序列分析软件进行分析,计算等位基因数目,鉴定单核苷酸多态性位点(single nuclear polymorphism,SNP).同时将序列导入Mega 4序列分析软件,计算错义突变率与同义突变率的比值(dN/dS),进行进化分析,构建进化树.结果 共抽提了60条日本血吸虫成虫的总RNA,其中46条通过RT-PCR获得了目的基因Sj-TSP2;序列分析共鉴定了20个等位基因,主要是由点突变引起,Sj-P2-10是

  5. 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

  6. Cloning and function analysis of high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Mainland strain)%日本血吸虫高速泳动家族B1蛋白基因的克隆表达与功能分析

    Institute of Scientific and Technical Information of China (English)

    姚媛; 许永良; 杨静; 余传信; 宋丽君; 殷旭仁; 王玠; 金一; 沈双; 张伟; 高玒

    2014-01-01

    Objective To clone and express a high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Main-land strain)and analyze its function. Methods The DNA fragment of open reading frame encoding Sj HMGB1 protein was ampli-fied by RT-PCR from the mRNA of S. japonicum worms,then it was subcloned into the expression vector pET28a(+)to form the recombinant expression plasmid SjHMGB1-pET28a. The recombinant expression plasmid was transformed into the component E. coli BL21(DE3),and the tranformant containing recombinant expression plasmid was induced with IPTG to express the recombi-nant protein SjHMGB1. The recombinant SjHMGB1 protein was purified by affinity chromatography with nickel chelating affinity chromatography agarose gel. The Gel retard experiment and animal immunization were performed to analyze the DNA binding ca- pacity and the immunologic property of recombinant SjHMGB1. The expression levels of HMGB1 in different life cycle stages of S. japonicum were analyzed by Western bloting and RT-PCR. Female ICR mice were immunized with the recombinant SjHMGB1 pro-tein and infected with 45±2 cercariae of S. japonicum after three immunizations. Forty-two days post-infection,the worms and eggs of S. japonicum were recovered from the portal vein and liver tissue,respectively. The worm and egg reduction rates were calculat-ed respectively. Results A 530 bp of specific DNA fragment was amplified from mRNA of S. japonicum by RT-PCR,which was the open reading frame(ORF)encoding SjHMGB1protein confirmed by DNA sequencing analysis. The recombinant expression plasmid SjHMGB1-pET28a was constructed by cloning the ORF of SjHMGB1 into a expression vector pET28a(+). The bacterium transformants containing the recombinant plasmid expressed a soluble recombinant protein about 28 kDa after induced by IPTG, and the recombinant SjHMGB1 protein was purified by nickel chelating affinity chromatography. The gel retard experiment showed that the recombinant SjHMGB1 protein could bind

  7. 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

  8. 鄱阳湖湖北钉螺日本血吸虫和外睾吸虫尾蚴逸出模式及其流行病学的初步调查%Preliminary investigation on the prevalence and cercarial rhythm pattern of Schistosoma japonicum and Exorchis sp.in Oncomelania hupensis (Gastropoda) populations in marshlands of Poyang Lake, China

    Institute of Scientific and Technical Information of China (English)

    Yassir Sulieman; Theerakamol Pengsakul; 黄帅钦; 彭午弦

    2013-01-01

    Digenean trematodes are a widespread group of parasitic flatworms that use snails somewhat exclusively as their first intermediate host and one species can competitively exclude other species in a single individual snail. In this study, prevalence of the trematodes, Schistosoma japonicum and Exorchis sp., among populations of Oncomelania hupensis and population density of the snail in four different marshland locations around Poyang Lake, China, was studied during spring (April) and autumn (October) of 2012, by using a 0.11 m2 frame sampler. Also, the rhythm of cercarial liberation of the two trematodes from this area was investigated. Results showed that the prevalence of Exorchis sp. infection among O. hupensis populations was high (2.01%), compared to S. japonicum (0.15%) over the four investigated sites during the spring- and autumn-collected samples. The mean density of live snails in spring was 12.2 per 0.11 m2 with overall infection of 0.3 snail per 0.11 m2;while in autumn, the mean density of live snails was 6.7 per 0.11 m2 with infection rate of 0.2 snail per 0.11 m2. Cercarial release pattern from S. japonicum was circadian, with a single peak of emergence between 12:00 and 14:00 pm, whereas that of Exorchis sp. displayed a bimodal peak of emergence: an early emergence peak between 06:00 and 08:00 am, and a late emergence peak between 16:00 and 18:00 pm. Finaly, the higher prevalence of Exorchis sp. infection among O. hupensis populations compared to S. japonicum at different sites and seasons in this study necessitates further investigation concerning interaction among trematode infections within O. hupensis populations, especially in regard to Exorchis sp., due to its importance in schistosomiasis biological control.%目的 复殖吸虫是一种广泛分布的寄生性扁形动物,它们专一性地利用贝类作为第一中间宿主.自然环境中常见到一种贝类是数种吸虫的中间宿主,但该贝类的每一个体通常只存在一

  9. 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

  10. Immuno-protection of SJIR-2 DNA vaccine with microspheres adjuvant in mice challenged with Schistosoma japonicum%日本血吸虫SJIR-2纳米微球核酸疫苗的免疫保护性研究

    Institute of Scientific and Technical Information of China (English)

    王正印; 潘丽红; 汪学龙

    2016-01-01

    目的:研究日本血吸虫胰岛素受体-2( SJIR-2)纳米微球核酸疫苗对小鼠攻击感染的免疫保护效果。方法构建pEGFP-SJIR-2重组质粒,双酶切鉴定并测序,大量提取pEGFP-SJIR-2质粒,用壳聚糖( CHS)修饰的聚乳酸-羟基乙酸共聚物( PLGA)微球包裹,用包裹后的SJIR-2纳米微球免疫小鼠。将40只雌性BALB/c小鼠随机分为4组( n=10),分别注射 PBS、空 pEGFP 质粒、CHS-PLGA 微球和 CHS-PL-GA-pEGFP-SJIR-2微球各100μg免疫小鼠,末次免疫2周后,用日本血吸虫尾蚴攻击感染小鼠,每次免疫及感染尾蚴前收集各组小鼠血清,ELISA法检测各组小鼠血清内免疫球蛋白( IgG)水平的变化。小鼠感染尾蚴42 d后全部剖杀,收集成虫和虫卵并计算减虫率和减卵率。结果成功构建了pEGFP-SJIR-2重组质粒,与PBS组比较, CHS-PLGA-pEGFP-SJIR-2组的成虫数和虫卵数差异有统计学意义( P<0.01)。CHS-PLGA-pEGFP-SJIR-2组的减虫率和减卵率分别为37.36%和46.82%,和PBS组相比,CHS-PLGA-pEGFP-SJIR-2组小鼠血清内IgG水平比明显增高( P<0.01),而pEGFP组和CHS-PLGA组成虫数和虫卵数与PBS组比较差异无统计学意义。结论 SJIR-2纳米微球核酸疫苗对感染血吸虫的BALB/c小鼠有一定的免疫保护效果,对其潜在的候选抗原疫苗的价值尚需深入研究。%Objective To research the immuno-protection of SJIR-2 DNA vaccine with nanometer microspheres a-gainst Schistosoma japonicum infection in mice. Methods To construct eukaryotic expression plasmid pEGFP-SJIR-2, identified by double digestion and sequenced delivery. The recombinant plasmid pEGFP-SJIR-2 was ex-tracted and was encapsulated into PLGA nanometer microspheres which were modified by CHS. 40 female BALB/c mice were randomly divided into 4 groups (n=10), each group of mice were injected with PBS, empty pEGFP plasmid, CHS-PLGA nanometer microspheres and CHS-PLGA-pEGFP-SJIR-2 nanometer microspheres 100 μg, re-spectively. Two weeks

  11. Survival of Oncomelania Snails from Lake Regions and Mountain Regions and Their Susceptibility to Schistosoma Japonicum under Simulated Field Conditions%模拟自然条件下湖沼型和山区型钉螺存活率及其血吸虫易感性研究

    Institute of Scientific and Technical Information of China (English)

    龚珍强; 李传昌; 汪雁; 黄重峰; 刘大仁; 陶波; 周宪民; 邹节新

    2014-01-01

    Objective To compare the 4-month survive rates of ribbed shell Oncomelania snails from lake regions of Xingzi County of Jiangxi Province,ribbed shell Oncomelania snails from mountain regions of Yushan County of Jiangxi Province and smooth shell Oncomelania snails from mountain regions of Pengshan County of Sichuan Province and their susceptibility to Schis-tosoma japonicum under simulated natural conditions.Methods Lake-region ribbed shell On-comelania snails,mountain-region ribbed shell Oncomelania snails and mountain-region smooth shell Oncomelania snails were collected from Xingzi County of Jiangxi Province,Yushan County of Jiangxi Province and Pengshan County of Sichuan Province,respectively.The infected and un-infected snails were screened using the cercariae escaping method.The Schistosoma japonicum-in-fected snails from Xingzi County were used to infect a rabbit.The liver was removed and miracidia were obtained after 45 days.Then snails were infected with the miracidia at a ratio of 120 at 26-30 ℃ under fluorescent lamp irradiation.These infected snails were fed for 4 months in Schistoso-miasis Control Station of Xingzi County under simulated natural conditions,and the survival rate and infection rate were determined.Results Oncomelania snails from mountain regions of Yushan County of Jiangxi Province and mountain regions of Pengshan County of Sichuan Province could survive under simulated natural conditions in Xingzi County of Jiangxi Province.But their survival rates were significantly lower than those from lake regions of Xingzi County of Jiangxi Province (P < 0.001 ).Furthermore,Oncomelania snails from lake regions of Xingzi County of Jiangxi Province and mountain regions of Yushan County of Jiangxi Province,but not those from moun-tain regions of Pengshan County of Sichuan Province,were easily infected with Schistosoma ja-ponicum miracidia from Xingzi County of Jiangxi Province.In addition,the infection rate of snails from lake regions of Xingzi

  12. 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

  13. 9个水稻栽培种和1个水稻组培种对稻螟赤眼蜂和赤眼卵蜂的互利作用%Synomonal effect of nine varieties and one culture of rice on Trichogramma japonicum Ashmead and Trichogramma chilonis (Ishii) (Hymenoptera: Trichogrammatidae)

    Institute of Scientific and Technical Information of China (English)

    ARCHNA; A. K. SINGH; A. V. N. PAUL; Anju JAIN

    2009-01-01

    采用培养皿法对营养生长期和开花期的9个水稻栽培种(Pusa Sugandh-2, Pusa Basmati-1, Pusa-2511, Pusa Basmati-370, Pusa-1077, Karnal Local, PRR-78, Jaya, Pusa-1238)和1个组培种(Culture No.34)的正己烷提取物进行了生测,以调查它们对稻螟赤眼蜂Trichogramma japonicum Ashmead和赤眼卵蜂Trichogramma chilonis (Ishii)平均寄生活性指数(PAI)和平均寄生百分率的影响.将不同水稻品种的正己烷提取物进行气相-液相色谱,来测定饱和碳氢化合物.其中,营养生长期的Pusa Sugandh-2叶片提取物激发两种寄生蜂产生最大的反应,平均寄生率最大.而开花期的Pusa Basmati-1使两种寄生蜂的平均寄生率最高.对营养生长阶段的叶片提取物进行的气相-液相色谱分析表明:Pusa Sugandh-2 含有25个碳原子(C25) 和29个碳原子 (C29)的2种化合物.对开花期的叶片提取物进行的气相-液相色谱分析也表明:Basmati-1 含有25个碳原子(C25)、26个碳原子(C26)和29个碳原子 (C29)的3种化合物,可激发寄生蜂产生最大的反应.%Bioassays of the hexane extracts of nine varieties viz., Pusa Sugandh-2, Pusa Basmati-1, Pusa-2511, Pusa Basmati-370, Pusa-1077, Karnal Local, PRR-78, Jaya, Pusa-1238 and one culture of rice, Culture No.34 in vegetative and flowering phase were carried out in Petri dish, to observe their synomonal effect on mean parasitoid activity index (PAI) and mean percent parasitism by Trichogramma japonicum, Ashmead and Trichogramma chilonis (Ishii). The hexane extracts of different rice varieties were subjected to gas liquid chromatography to determine the saturated hydrocarbons. Among these, leaf extracts from variety Pusa Sugandh-2 elicited maximum response in vegetative phase for both the parasitoids by way of mean percent parasitism. Whereas in flowering phase variety Pusa Basmati-1 elicited maximum mean percent parasitism. Gas liquid chromatography of the leaf extracts from vegetative phase revealed that Pusa

  14. Molecular diversity and phylogeny of rhizobia associated with Lablab purpureus (Linn.) grown in Southern China.

    Science.gov (United States)

    Chang, Yue Li; Wang, En Tao; Sui, Xin Hua; Zhang, Xiao Xia; Chen, Wen Xin

    2011-06-01

    As an introduced plant, Lablab purpureus serves as a vegetable, herbal medicine, forage and green manure in China. In order to investigate the diversity of rhizobia associated with this plant, a total of 49 rhizobial strains isolated from ten provinces of Southern China were analyzed in the present study with restriction fragment length polymorphism and/or sequence analyses of housekeeping genes (16S rRNA, IGS, atpD, glnII and recA) and symbiotic genes (nifH and nodC). The results defined the L. purpureus rhizobia as 24 IGS-types within 15 rrs-IGS clusters or genomic species belonging to Bradyrhizobium, Rhizobium, Ensifer (synonym of Sinorhizobium) and Mesorhizobium. Bradyrhizobium spp. (81.6%) were the most abundant isolates, half of which were B. elkanii. Most of these rhizobia induced nodules on L. purpureus, but symbiotic genes were only amplified from the Bradyrhizobium and Rhizobium leguminosarum strains. The nodC and nifH phylogenetic trees defined five lineages corresponding to B. yuanmingense, B. japonicum, B. elkanii, B. jicamae and R. leguminosarum. The coherence of housekeeping and symbiotic gene phylogenies demonstrated that the symbiotic genes of the Lablab rhizobia were maintained mainly through vertical transfer. However, a putative lateral transfer of symbiotic genes was found in the B. liaoningense strain. The results in the present study clearly revealed that L. purpureus was a promiscuous host that formed nodules with diverse rhizobia, mainly Bradyrhizobium species, harboring different symbiotic genes.

  15. 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

  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. The Rhizobium etli rpoN Locus: DNA Sequence Analysis and Phenotypical Characterization of rpoN, ptsN, and ptsA Mutants

    OpenAIRE

    Michiels, Jan; Van Soom, Tom; D’hooghe, Inge; Dombrecht, Bruno; Benhassine, Traki; Wilde, Petra; Vanderleyden, Jos

    1998-01-01

    The rpoN region of Rhizobium etli was isolated by using the Bradyrhizobium japonicum rpoN1 gene as a probe. Nucleotide sequence analysis of a 5,600-bp DNA fragment of this region revealed the presence of four complete open reading frames (ORFs), ORF258, rpoN, ORF191, and ptsN, coding for proteins of 258, 520, 191, and 154 amino acids, respectively. The gene product of ORF258 is homologous to members of the ATP-binding cassette-type permeases. ORF191 and ptsN are homologous to conserved ORFs f...

  18. Rhizobium strains differ considerably in outer membrane permeability and polymyxin B resistance.

    Science.gov (United States)

    Komaniecka, Iwona; Zamłyńska, Katarzyna; Zan, Radosław; Staszczak, Magdalena; Pawelec, Jarosław; Seta, Irena; Choma, Adam

    2016-01-01

    Six rhizobium (Rhizobium leguminosarum bv. Trifolii TA1, Sinorhizobium meliloti 1021, Mesorhizobium huakuii IFO 15243(T), Ochrobactrum lupini LUP 21(T), Bradyrhizobium japonicum USDA110 and B. elkanii USDA 76) and two Escherichia coli strains (E. coli ATCC 25922 and E. coli HB 101) were compared in respect to polymyxin B and EDTA resistance, as well as bacterial outer membrane (OM) permeability to a fluorescent hydrophobic agent (N-phenyl-1-naphthylamine - NPN). TEM (Transmission Electron Microscopy) and a microbial test demonstrated that all the rhizobia were much more resistant to polymyxin B in comparison with E. coli strains. EDTA and polymyxin B enhance permeability of B. japonicum and O. lupini OM. Other rhizobia incorporated NPN independently of the presence of membrane-deteriorating agents; however, the level of fluorescence (measured as NPN absorption) was strain dependent.

  19. 血吸虫病肝纤维化小鼠IL-22表达及肝星状细胞衰老的意义%Dynamic expressions of IL-22 and hepatic stellate cells senescence in mice in-fected with Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    李兰; 孙颖; 王洪武; 黄宇; 黄加权; 宁琴

    2014-01-01

    Objective To investigate the dynamic expressions of interleukin-22(IL-22),Interleukin-22 receptor 1(IL-22R1),and hepatic stellate cells(HSC)senescence in mice with Schistosoma japonicum infection. Methods A murine model of S. japonicum infection was established and the serum samples and liver tissues were collected 4,6,8,12 weeks post-infection. The serum samples were detected for the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST). The pathological changes and proliferation of hepatic collagen fibers in the liver tissue were observed after HE staining and Masson staining. The HSC senescence in fibrotic livers was determined by the detection of senescence-associatedβ-galactosidase(SA-β-Gal). Sandwich ELISA was used to measure the expressions of IL-22,and Real-time PCR was used to test the mRNA levels of IL-22 and IL-22R1. The control group without S. japonicum infection was set up. Results The serum levels of ALT and AST signifi-cantly increased 8 weeks and 12 weeks after the infection(vs. 0 week,all P<0.05). The level of IL-22 increased 4 weeks and 6 weeks after the infection(vs. 0 week,both P<0.05),but reduced 8 weeks post-infection,and was even lower 12 weeks post-in-fection(vs. 4 weeks and 6 weeks,both P<0.01). Being consistent with the dynamic expression of IL-22 protein,the mRNA ex-pression of IL-22 began to increase 4 weeks and reached the peak 6 weeks after the infection(vs. 0 week,both P<0.05),and continuously declined 8 weeks and 12 weeks post-infections(vs. 6 weeks,both P<0.05). The increase of the expression of IL-22R1 mRNA was correlated with the progression of fibrosis,and the peak was in 12 weeks post-infections(vs. 0 week and 6 weeks,both P<0.05). The number of senescence-associated beta-galactosidase-positive HSCs was reduced with the decreasing expression of IL-22 in the advanced liver fibrosis. Conclusion IL-22 and IL-22R1 are involved in the pathogenesis of schistoso-miasis liver fibrosis. As an inflammation factor

  20. 山蚂蝗慢生根瘤菌的遗传多样性及系统发育%Genetic diversity and phylogeny of bradyrhizobia associated with Desmodium spp

    Institute of Scientific and Technical Information of China (English)

    谷峻; 张静苗; 贾瑞宗; 陈文新

    2011-01-01

    [目的]研究我国亚热带和温带地区与山蚂蝗共生的慢生根瘤菌遗传多样性和系统发育.[方法]采用BOX-PCR和多位点基因序列分析(nifH,nodC和recA基因)方法对分离自我国不同地区的29株山蚂蝗慢生根瘤菌进行遗传多样性和系统发育分析.[结果]BOX-PCR分析表明供试的山蚂蝗慢生根瘤菌形成25个基因遗传型,具有丰富的基因组多样性.多位点基因序列分析发现代表菌株位于慢生根瘤菌的3个分支上,分别与埃氏慢生根瘤菌(Bradyrhizobium elkanii),大豆慢生根瘤菌(Bradyrhizobium japonicum)和圆明慢生根瘤菌(Bradyrhizobium yuanmingense)亲缘关系近.[结论]我国山蚂蝗慢生根瘤菌具有丰富的遗传多样性,共生基因系统发育分析表明它们多与持家基因共同进化,并以垂直进化为主.%[Objective ] Genetic diversity and phylogeny of bradyrhizobial strains associated with Desmodium spp. In subtropic and temperate regions of China were analyzed. [ Methods ] We studied 29 desmodia isolates from different regions with BOX-PCR fingerprinting and multilocus sequence analysis ( nifH, nodC and recA gene) to describe the genotypic characteristics and phylogenetic relationships. [Results] We achieved 25 genotypes with BOX-PCR genomic fingerprinting analysis, indicating that the tested strains had a great diversity at genomic level. The representative bradyrhizobial strains were located in three phylogenic branches with multilocus sequence analysis ( nifH, nodC and recA gene) , that was closely related to Bradyrhizobium elkanii, Bradyrhizobium japonicum and Bradyrhizobium yuanmingense, respectively. [ Conclusion] The desmodia bradyrhizobia had abundantly diversity. Diverse symbiotic genes including nifH and nodC genes were also found in these strains that indicated that the symbiotic genes were mainly maintained by vertical transfer in these bradyrhizobial populations and coevolved with housekeeping genes.

  1. EXPRESSION OF SCHISTOSOMA JAPONICUM CALRETICULIN PROTEIN IN SF9 INSECT CELLS AND PRELIMINARY ANALYSIS OF ITS ROLE IN THE ACTIVATION OF DENDRITIC CELLS%日本血吸虫钙网织蛋白在Sf9昆虫细胞中的表达及其活化树突细胞的初步分析

    Institute of Scientific and Technical Information of China (English)

    李莹; 李运燕; 徐瑞敏; 苑纯秀; 杨健美; 林矫矫; 李利珍; 冯新港

    2012-01-01

    为获得在Sf9昆虫细胞中表达的日本血吸虫钙网织蛋白(Schistosoma japonicum calreticulin protein,SjCRT)并分析其活化小鼠骨髓来源树突状细胞的功能,将构建的重组杆状病毒转移载体pFastBacHTA-SjCRT转入DH 10Bac细胞,得到重组穿梭质粒reBacmid-SjCRT,再转染到Sf9昆虫细胞,进行重组蛋白的表达.用Westem blot和间接免疫荧光对表达蛋白进行鉴定.His柱亲和层析法纯化表达的蛋白,Westen blot鉴定纯化后的蛋白.从BALB/c小鼠产生骨髓来源的树突细胞mDCs,用纯化的重组SjCRT蛋白与mDCs共培养,流式细胞术检测mDCs细胞的表面分子MHCⅡ、CD40和CD86的表达.结果显示,在Sf9昆虫细胞中成功表达了SjCRT蛋白;纯化后的重组SjCRT蛋白既能被感染日本血吸虫42 d的兔阳性血清识别,也能被原核表达的重组SJC RT蛋白免疫鼠的血清所识别.流式细胞术结果显示,与对照组的相比,SjCRT蛋白刺激组mDCs细胞表面分子MHCⅡ和CD86的表达量显著增强(P<0.05).可见,在Sf9昆虫细胞中表达的SjCRT蛋白能刺激小鼠骨髓来源树突细胞表型的成熟.

  2. 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.

  3. 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

  4. Recombinant Plasmid ZLW/pEGFP-C2 Transfection into Schistosomula of Schistosoma japonicum%日本血吸虫表膜特异结合肽重组质粒ZLW/pEGFP-C2体外转染日本血吸虫童虫的研究

    Institute of Scientific and Technical Information of China (English)

    刘彦; 曾庆仁; 杨胜辉; 魏琦; 周军; 李立新; 兰玲梅

    2011-01-01

    目的 观察日本血吸虫表膜特异结合肽合成基因(ZLW)构建的ZLW/pEGFP-C2重组质粒体外转染H本血吸虫童虫的效率,了解其抗虫作用.方法 用0.75%二甲基亚砜(DMSO)和高浓度质粒浸泡法,使用重组质粒ZLW/pEGFP-C2转染机械转化的日本血吸虫脱尾童虫,以瞬时表达的增强型绿色荧光蛋白(EGFP)基因为报告基因,在倒置荧光镜下观察虫体.抽提转染后培养48 h虫体的总RNA和全虫蛋白,分别用RT-PCR和蛋白质印迹(Western blotting)检测ZLW基因和EGFP基因在虫体内的表达情况.于转染后培养24、48、72和96 h,用美蓝染色法鉴别虫体存活情况,并计数.以上试验均设空质粒组和Tris-HCl缓冲盐溶液(TBS)对照组.结果 镜下观察结果显示,转染率约为10%,其EGFP主要分布于虫体皮层,以口、腹吸盘最为明显;空质粒转染组虫体腹吸盘处略带绿色荧光.RT-PCR结果显示,ZLW/pEGFP-C2转染组虫体的扩增产物大小约为259 bp,测序结果与ZLW序列一致.Western blotting分析证实EGFP基因在童虫体内表达.抗虫效果显示,转染后24 h和48 h,ZLW/pEGFP-C2转染组童虫死亡率(14.0%,48.8%)与空质粒组(15.9%,45.7%)和TBS对照组(16.9%,50.3%)间的差异无统计学意义(P>0.05);转染后72 h,ZLW/pEGFP-C2转染组死亡率(92.7%)高于空质粒组(73.2%)和TBS对照组(76.8%)(P<0.01);转染后96 h,ZLW/pEGFP-C2转染组童虫死亡率达100%.结论 DMSO作用的高浓度质粒浸泡法可将重组质粒ZLW/pEGFP-C2引入日本血吸虫章虫体内,并获得表达.%Objective To study the efficiency of ZLW/pEGFP-C2 plasmid transfection into Schistosoma japonicum schistosomula and observe its in vitro effect of anti-schistosomula. Methods Recombinant plasmid ZLW/pEGFP-C2 was transfected into mechanically transformed schistosomula by immersion in 0.75% DMSO and high concentration of plasmid. Enhanced green fluorescent protein (EGFP) transfected cells were observed under inverted fluorescence

  5. 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

  6. 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.

  7. Phylogenetic relationship of Lotus uliginosus symbionts with bradyrhizobia nodulating genistoid legumes.

    Science.gov (United States)

    Lorite, María J; Videira e Castro, Isabel; Muñoz, Socorro; Sanjuán, Juan

    2012-02-01

    Lotus species are legumes with potential for pastures in soils with low-fertility and environmental constraints. The aim of this work was to characterize bacteria that establish efficient nitrogen-fixing symbiosis with the forage species Lotus uliginosus. A total of 39 isolates were obtained from nodules of L. uliginosus naturally growing in two different locations of Portugal. Molecular identification of the isolates plus the commercial inoculant strain NZP2039 was performed by REP-PCR, 16S rRNA RFLP, and 16S rRNA, glnII and recA sequence analyses. Limited genetic diversity was found among the L. uliginosus symbionts, which showed a close phylogenetic relationship with the species Bradyrhizobium japonicum. The symbiotic nifH, nodA and nodC gene sequences were closely related with the corresponding genes of various Bradyrhizobium strains isolated from Lupinus and other genistoid legumes and therefore were phylogenetically separated from other Lotus spp. rhizobia. The L. uliginosus bradyrhizobia were able to nodulate and fix nitrogen in association with L. uliginosus, could nodulate Lotus corniculatus with generally poor nitrogen-fixing efficiency, formed nonfixing nodules in Lotus tenuis and Lupinus luteus roots and were unable to nodulate Glycine soja or Glycine max. Thus, L. uliginosus rhizobia seem closely related to B. japonicum biovar genistearum strains.

  8. Phenotype comparison between bone marrow derived dendritic cell and DC2.4 cell stimulated with antigen from Schistosoma japonicum%血吸虫抗原刺激小鼠骨髓来源的树突状细胞与DC2.4细胞的表型比较

    Institute of Scientific and Technical Information of China (English)

    李小红; 曹建平; 汤林华; 王胜军; 成静

    2011-01-01

    目的 研究比较小鼠树突状细胞DC2.4和骨髓来源树突状细胞(bone marrow derived dendritic cell,BMDC)经血吸虫抗原谷胱甘肽转移酶(GST)刺激后表面分子的表达异同.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 colonystimulating factor,GM-CSF)诱导培养,获得树突状细胞.常规方法培养DC2.4.体外用日本血吸虫抗原GST刺激前述两种细胞,以PBS和脂多糖(lipopolysaccharide,LPS)作对照,流式细胞仪检测细胞表面分子CD40、CDSO、CD86的平均荧光强度,并进行统计学分析.结果 日本血吸虫抗原GST刺激BMDC后,表面分子CD40、CD80、CD86的平均荧光强度依次为100.39、42.38、170.83,与PBS对照组比较,CD40无明显变化,而CD80、CD86表达上调(P<0.01);GST刺激DC2.4后,细胞表面分子CD40、CD80、CD86的平均荧光强度依次为23.73、72.13、59.58,与PBS对照组比较,CD40和CD86表达上调(P<0.01),而CD80变化不明显.结论 DC2.4与BMDC经日本血吸虫抗原刺激后表面分子的表达变化不同.%Objective To compare the phenotypes of bone marrow derived dendritic cell(BMDC)and DC2.4 cell stimulated with GST from Schistosoma japonicum.Methods Bone marrow cells were cultured in the presence of IL-4 and GM-CSF to induce dendritic cells.DC2.4 cells were cultured as routine.Both cells were stimulated with GST and the expressions of CD40,CD80 and CD86 on the cells'surface were analyzed by FACS,using PBS and lipopolysaccharide as controls. Results After stimulating with GST,the means of fluorescence intensity(MFI)for CD40,CD80 and CD86 on BMDC surface were 100.39,42.38 and 170.83,respectively.Compared with PBS control,the MFI of CD80 and CD86 on BMDC,but not CD40,enhanced significantly.The MFIs of CD40.CD80 and CD86 on DC2.4 loaded by GST were 23.73,72.13 and 59.58 respectively.Compared with PBS control,the expressions of CD40 and CD86 enhanced significantly after schistosome

  9. Preliminary Observation on the Effect of Mefolquine against Egg Granuloma Formation in the Liver of Mice Infected with Schistosoma japonicum%甲氟喹对感染小鼠日本血吸虫虫卵肉芽肿形成影响的初步观察

    Institute of Scientific and Technical Information of China (English)

    肖树华; 张超威

    2013-01-01

    形成具有抑制作用.%Objective To explore whether mefolquine possesses the effect on granuloma formation induced by Schistosoma japonicum eggs. Methods Seventeen out of twenty-eight mice infected with 20 S. japonicum cercariae for 35 days were treated orally with mefloquine at a single dose of 200 mg/kg, and groups of 2-3 mice were sacrificed at various intervals post-treatment. The livers removed from each group of mice were fixed in 10% formaldhyde. While the remained 11 untreated mice divided into 6 groups (1-2 mice per group) were sacrificed at the same time periods as groups of mice treated with mefloquine, and their livers served as untreated corresponding controls. The granulomas with single egg in the center were counted and their diameters were measured using an ocular micrometer. The liver tissue sections were stained with hematoxylin and eosin (HE), Foot's or Mallory's methods for observation on histopathological alteration of egg granulomas, and on the appearance of reticular and collagen fibers within the granulomas. Results After infected mice were treated with mefloquine for 3, 7, 14, 21, 28 and 35 days, i.e., 38, 42, 49, 56, 63, and 70 days post-infection, the mean diameters of granuloma with single egg measured in the liver tissues section were (161 ±19), (175±13), (195±9), (171±40), (180±13), and (145±25)μm, respectively, and each of them was significantly lower than that of its corresponding control group of (189±18), (197±11), (211±12), (208±19), (203±16), and (207±36) μm (P<0.01 or P<0.05). Histopatological observation showed that in mice treated with mefloquine, the eosinophil-predominant inflammatory cells around the egg granuloma were sustained to 14-21 d post treatment (49-56 d post infection), which was significantly different from the corresponding control groups that all the eggs were surrounded by fibroblasts at 42 d post infection. Up to 28-35 d post treatment (63-70 d post infection), the boundary of egg granulomas distributed in the liver tissues of

  10. Genetic diversity of indigenous soybean-nodulating rhizobia in response to locally-based long term fertilization in a Mollisol of Northeast China.

    Science.gov (United States)

    Yan, Jun; Chen, WenFeng; Han, XiaoZeng; Wang, EnTao; Zou, WenXiu; Zhang, ZhiMing

    2017-01-01

    The influences of five different fertilizer treatments on diversity of rhizobia in soybean nodule were investigated in a long-term experiment with with four replicates: (1) control (without fertilization), (2) balanced NPK fertilizer (NPK), and (3-5) unbalanced chemical fertilizers without one of the major elements (NP, PK, and NK) in Mollisol in Northeast China. The highest soybean yield was observed in the NPK treatment. Total of 200 isolates were isolated and grouped into four Bradyrhizobium genospecies corresponding to B. japonicum, B. diazoefficiens, B. ottawaense and Bradyrhizobium sp. I, based upon the multilocus sequence analysis of 6 housekeeping genes. The Bradyrhizobium sp. I was extensively distributed throughout the study site and was recorded as the dominant soybean rhizobia (82.5-87.5%). Except the NK treatment, the other fertilizer treatments had no effect on rhizobial species composition. Compared with the CK treatment, all the fertilizer treatments decreased species richness, diversity and evenness. The soil organic carbon contents, available N content and pH were the key soil factors to rhizobial community structure. Results suggest that long-term fertilization can decrease rhizobial species diversity, while balanced fertilization with NPK is the most suitable fertilization regime if taking both soybean yields and rhizobial diversity into account.

  11. Characterization of soybean rhizobia at different levels using PCR based techniques%采用PCR-RFLP技术在不同水平上鉴定大豆根瘤菌

    Institute of Scientific and Technical Information of China (English)

    张学贤; M.木论贝格; 李德安; 周俊初; 李阜棣

    1999-01-01

    Nineteen standard USDA strains representing three recognized soybean rhizobia, Sinorhizobium fredii,Bradyrhizobium japonicum and B. elkanii, were examined by restriction fragment length polymorphism (RFLP)analysis of 16S rRNA genes amplified by polymerase chain reaction (PCR). Four composite genotypes were obtained from the combined data of the RFLP analysis with three endonucleases. S. fredii, B. japonicum, B. elkanii Ⅱ and Ⅱa reference strains fell into these four genotypes respectively. No genotype was shared by two species. Therefore,16S rDNA PCR-RFLP is a rapid tool for the identification of soybean rhizobia. According to 16S rDNA PCRRFLP, 22 fast-growing and 19 slow-growing soybean rhizobia strains from China were identified to S. fredii and B . japonicum , respectively.The reference strains and wild type isolates were also examined by 16S~23S intergenic spacer PCR-RFLP analysis. PCR results indicated that the length of the intergenic region between S. fredii and Bradyrhizobium sp.is different. All fast-growing S. fredii strains produced one larger band (2.1 kb), and all slow-growing Bradyrhizobium strains produced one smaller band (2.0 kb). Based on 16S~23S RFLP analysis, 22 S. fredii isolates can be further separated into two "genotypes" that originated from two different regions in China; 19 B. japonicum strains from Heilongjiang Province (northeast of China) can be separated into 11 genotypes. These wild type isolates were also characterized at strain level by REP (repetitive extragenic palindromic) and ERIC (enterobacterial repetitive intergenic consensus) PCR fingerprinting.%采用16S rRNA基因PCR扩增与限制性酶切片段多态性分析(RFLP)技术对选自弗氏中华根瘤菌(S.fredii)、大豆慢生根瘤菌(B.japonicum)和埃氏慢生根瘤菌(B.elkanii)的19株代表菌进行了比较分析,根据用3种限制性内切酶的RFLP分析结果,可将供试菌株分为S.fredii,B.japonicum,B.elkaniiⅡ和B.elkaniiⅡa等4种基因型.各

  12. 日本血吸虫重组质粒pGEX-Sj26GST的构建及在大肠埃希菌BL21(DE3)中的表达%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; 李文桂; 向进平

    2013-01-01

    目的 构建日本血吸虫(Schistosoma japonic um,Sj)重组质粒pGEX-Sj26GST,并研究其在大肠埃希菌(Escheric hia coli,E.coli BL21 (DE3)中的表达.方法 使用RNeasy Mini试剂盒提取sj成虫总RNA,实时荧光定量PCR(real-time PCR,RT-PCR)扩增日本血吸虫26 kDa的谷胱甘肽-S-转移酶(Sj26GST)编码基因,并将扩增产物定向克隆到pGEX-1λT载体上,构建重组质粒pGEX-Sj26GST;将重组质粒电穿孔转化感受态大肠埃希菌BL21(DE3),异丙基硫代-β-D-半乳糖苷(IPTG)诱导其表达;通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表达产物及蛋白印迹(Western blot)试验鉴定表达的融合蛋白的抗原性.结果 RT-PCR扩增出676 bp大小的Sj26GST抗原编码基因;双酶切试验证实Sj26GST抗原编码基因成功克隆到pGEX-1λT载体中;SDS-PAGE分析出现相对分子质量(Mr)约52×103的融合蛋白,薄层扫描分析显示表达的融合蛋白约占总菌体蛋白的20%,Western blot结果显示融合蛋白可被Sj感染的兔血清识别.结论 成功构建重组质粒pGEX-Sj26GST,其在大肠埃希菌BL21(DE3)中可获得表达,且表达的融合蛋白具有特异的抗原性.%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 Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double

  13. 日本血吸虫间接血凝法抗体检测试剂盒稀释液的改进%Alternative diluent used for the commercial IHA assay kit for detection of Schistosoma japonicum anti-body

    Institute of Scientific and Technical Information of China (English)

    章乐生; 王恩木; 张世清; 汪奇志; 呼明闯; 司武敏; 杨荣笙

    2014-01-01

    目的:确定一种适用于安徽安吉医药科技有限公司新包装试剂盒的稀释液,替代原包装中的冻干致敏红细胞稀释液(生理盐水)和血清标本稀释液(蒸馏水)。方法以pH7.2磷酸盐缓冲液作为稀释液制备新包装试剂盒替代原包装,比较新包装试剂盒与原包装的敏感性和特异性以及与肝吸虫、肺吸虫、姜片虫和钩虫等病人血清交叉反应情况;比较各试剂盒的最低检出量,观察凝集反应模式。结果新包装试剂盒与原包装敏感性分别为95%和94.2%,差异无统计学意义(χ2=0.000,P=1.000);特异性分别为96.9%和95.8%,差异无统计学意义(χ2=0.000,P=1.000);约登指数分别为0.92和0.9;最低检出量均为1:1280,磷酸盐缓冲液组凝集模式较好;与肝吸虫、姜片虫和钩虫病人的血清交叉反应率均为0,与肺吸虫的交叉反应率均为19.4%。结论确定磷酸盐缓冲液作为日本血吸虫间接血凝法抗体检测试剂盒新包装的稀释液,替代原包装中的致敏红细胞稀释液和标本稀释液。%Objective To evaluate the feasibility of a new diluent as a substitute for the original ones (sa⁃line used for diluting the lyophilized sensitized red cells, and distilled water for diluting the serum specimen) packaged in the commercial indirect hemagglutination (IHA)assay kit manufactured by Anhui Anji Pharmaceu⁃tical Technology Co., Ltd. Methods Phosphate buffered solution(PBS, pH7.2)was used to replace the diluent in the original kit, and two preparations were compared regarding the sensitivity and specificity in detection of Schistosoma japonicum as well as the cross-reactivity with liver fluke, lung fluke, giant intestinal fluke and hook⁃worm. The minimal quantification and the agglutination pattern in each group were also observed. Results The sensitivity and specificity were 95% and 96.9% for the modified kit, 94.2% and 95.8% for the original one

  14. The inhibitory effect of eupatorium japonicum thunb and foeniculum vulgare extract on prostatic hyperpla-sia in rats%泽兰与小茴香提取物抑制大鼠前列腺增生的实验研究

    Institute of Scientific and Technical Information of China (English)

    唐维; 代光成; 薛波新; 单玉喜; 张文芳

    2014-01-01

    能显著抑制大鼠前列腺增生,其作用机制在一定程度上与降低大鼠血清睾酮、双氢睾酮含量及睾酮/雌二醇值有关。%Objective Benign prostatic hyperplasia ( BPH) is one of common diseases in aged males , and searching for new therapeutic drugs to BPH has been a research hotspot in recent years .This article was to study the inhibitory effect of eupatorium ja-ponicum thunb and foeniculum vulgare extract ( EFE) on benign prostatic hyperplasia in rats and its possible mechanism . Methods 48 male rats were randomly divided into 6 groups:normal control group without any treatment , model group of BPH treated with subcu-taneous injection of testosterone propionate , positive control group of BPH treated with dutasteride , high, middle and low dosage groups according to different EFE dosage (156 mg/kg, 234 mg/kg and 312 mg/kg).45 days after the treatment, the rats were sacrificed for measurement of the prostate glandular wet weight , the index of prostate gland ( PI ) , the morphological changes of prostate gland by light microscopy and the content of sex hormone . Results The prostate wet weight and PI decreased after EFE treatment for 45 days compared with the BPH model group(P<0.01 ).The hyperplastic glandular epithelium papilla waned and even disappeared in three EFE groups under the light microscope , and the epithelial cells became cubical or flat .High dosage EFE group (312 mg/kg) has simi-lar efficacy to dutasteride group .EFE significantly reduced serum testosterone content , dihydrotestosterone content and T/E2 ratio( P<0.05 ). Conclusion EFE can significantly inhibit prostatic hyperplasia in rats , and its mechanism is related to the decrease of the contents of serum testosterone and dihydrotestosterone as well as T/E2 ratio.

  15. 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

  16. Effect of bacterial root symbiosis and urea as source of nitrogen on performance of soybean plants grown hydroponically for bioregenerative life support systems (BLSSs

    Directory of Open Access Journals (Sweden)

    Roberta eParadiso

    2015-10-01

    Full Text Available Soybean is traditionally grown in soil, where root symbiosis with Bradyrhizobium japonicum can supply nitrogen (N, by means of bacterial fixation of atmospheric N2.Nitrogen fertilizers inhibit N-fixing bacteria. However urea is profitably used in soybean cultivation in soil, where urease enzymes of telluric microbes catalyze the hydrolysis to ammonium, which has a lighter inhibitory effect compared to nitrate.Previous researches demonstrated that soybean can be grown hydroponically with recirculating complete nitrate-based nutrient solutions. In Space, urea derived from crew urine could be used as N source, with positive effects in resource procurement and waste recycling. However, whether the plants are able to use urea as the sole source of N and its effect on root symbiosis with B. japonicum is still unclear in hydroponics.We compared the effect of two N sources, nitrate and urea, on plant growth and physiology, and seed yield and quality of soybean grown in closed-loop Nutrient Film Technique (NFT in growth chamber, with or without inoculation with B. japonicum.Urea limited plant growth and seed yield compared to nitrate by determining nutrient deficiency, due to its low utilization efficiency in the early developmental stages, and reduced nutrients uptake (K, Ca and Mg throughout the whole growing cycle. Root inoculation with B. japonicum did not improve plant performance, regardless of the N source. Specifically, nodulation increased under fertigation with urea compared to nitrate, but this effect did not result in higher leaf N content and better biomass and seed production.Urea was not suitable as sole N source for soybean in closed-loop NFT. However the ability to use urea increased from young to adult plants, suggesting the possibility to apply it during reproductive phase or in combination with nitrate in earlier developmental stages. Root symbiosis did not contribute significantly to N nutrition and did not enhance the plant ability

  17. Rhizosphere effect of colonizer plant species on the development of soil microbial community during primary succession on postmining sites

    Energy Technology Data Exchange (ETDEWEB)

    Elhottova, D.; Kristufek, V.; Maly, S.; Frouz, J. [Academy of Sciences of the Czech Republic, Ceske Budejovice (Czech Republic). Inst. for Soil Biology

    2009-07-01

    The impact of pioneer plant species Tussilago farfara on structural, functional, and growth characterization of microbial community colonizing the spoil colliery substrate was studied in a laboratory microcosm experiment. Microcosms consisting of spoil substrate (0.7 dm{sup 3} of tertiary alkaline clay sediment from Sokolov brown-coal mine area) from a pioneer site (without vegetation, 5 years after heaping) were cultivated in a greenhouse with one plant of this species. Plant roots substantially increased microbial diversity and biomass after one season (7 months) of cultivation. Roots influenced the microbial community and had nearly twice the size, higher growth, and metabolic potential in comparison to the control. The development of microbial specialists improves the plant nutrient status. Bacterial nitrogen (N{sub 2}) fixators (Bradyrhizobium japonicum, Rhizobium radiobacter) and arbuscular mycorrhizal fungi were confirmed in the rhizosphere of Tussilago farfara.

  18. Efficient synthesis of the intermediate of abacavir and carbovir using a novel (+)-γ-lactamase as a catalyst.

    Science.gov (United States)

    Gao, Shuaihua; Zhu, Shaozhou; Huang, Rong; Lu, Yingxiu; Zheng, Guojun

    2015-09-15

    The enantiomers of 2-azabicyclo[2.2.1]hept-5-en-3-one (γ-lactam) are key chiral synthons in the synthesis of antiviral drugs such as carbovir and abacavir. (+)-γ-Lactamase can be used as a catalyst in the enzymatic preparation of optically pure (-)-γ-lactam. Here, a (+)-γ-lactamase discovered from Bradyrhizobium japonicum USDA 6 by sequence-structure guided genome mining was cloned, purified and characterized. The enzyme possesses a significant catalytic activity towards γ-lactam. The active site of the (+)-γ-lactamase was studied by homologous modeling and molecular docking, and the accuracy of the prediction was confirmed by site-specific mutagenesis. The (+)-γ-lactamase reveals the great practical potential as an enzymatic method for the efficient production of carbocyclic nucleosides of pharmaceutical interest.

  19. 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.

  20. Avaliação de populações de possíveis rizobactérias em solos sob espécies florestais Evaluation of possible rhizobacteria populations in soils under forest species

    Directory of Open Access Journals (Sweden)

    Rodrigo Matheus Pereira

    2008-10-01

    Full Text Available Embora estudos recentes relatem a utilização de RCPC (Rizobactérias Promotoras de Crescimento de Plantas no Brasil, raríssimos trabalhos avaliam a presença natural dessas espécies bacterianas no solo. O objetivo deste trabalho foi avaliar a ocorrência de RPCP em duas amostras de solo sob diferentes tipos de manejo, através da construção e do seqüenciamento de bibliotecas de DNA metagenômico. Utilizaram-se oligonucleotídeos específicos para amplificação da região hipervariável do espaço intergênico dos genes ribossomais 16S-23S de DNA extraído de diferentes solos, sob Eucalyptus sp. e sob mata. Os fragmentos obtidos foram inseridos em vetor e clonados. As bibliotecas geraram 495 clones, que foram seqüenciados e identificados através de comparações realizadas pelo software Blast. O solo sob Eucalyptus sp. apresentou maior número de RPCP do que sob mata. Os filos Actinobacteria e Proteobacteria eram maiores no solo sob Eucalyptus sp., estando o filo Firmicutes ausente no solo sob mata. Somente oito espécies diferentes de RPCP foram detectadas: Bacillus subtilis, Bacillus megaterium, Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium sp., Frankia sp., Pseudomonas fluorescens e Pseudomonas gladioli. O trabalho forneceu valiosos dados sobre a presença de RPCP em solos com espécies florestais e sua possível utilização em reflorestamentos, assim como para o melhor conhecimento desses microrganismos nos solos do Brasil.Although new studies describe the use of PGPR (Plant Growth-Promoting Rhizobacteria in Brazil, they rarely evaluate the natural existence of these bacterial species in the soil. The objective of this study was to evaluate PGPR in two samples under different use types, one with native forest and the other with eucalyptus, through construction and sequencing of a metagenomic DNA library. Using specific probes from the internally transcribed region of 16S-23S rRNA genes, fragments of PCR products

  1. A lipochito-oligosaccharide, Nod factor, induces transient calcium influx in soybean suspension-cultured cells.

    Science.gov (United States)

    Yokoyama, T; Kobayashi, N; Kouchi, H; Minamisawa, K; Kaku, H; Tsuchiya, K

    2000-04-01

    Lipochito-oligosaccharides (Nod factors) produced by Rhizobium or Bradyrhizobium are the key signal molecules for eliciting nodulation in their corresponding host legumes. To elucidate the signal transduction events mediated by Nod factors, we investigated the effects of Nod factors on the cytosolic [Ca2+] of protoplasts prepared from roots and suspension-cultured cells of soybean (Glycine max and G. soja) using a fluorescent Ca2+ indicator, Fura-PE3. NodBj-V (C18:1, MeFuc), which is a major component of Nod factors produced by Bradyrhizobium japonicum, induces transient elevation of cytosolic [Ca2+] in the cells of soybean within a few minutes. This effect is specific to soybean cells and was not observed in the tobacco BY-2 cells. Furthermore, NodBj-V without MeFuc did not induce any cytosolic [Ca2+] elevation in soybean cells. Exclusion of Ca2+ from the medium, as well as pre-treatment of the cells with an external Ca2+ chelator or with a plasma membrane voltage-dependent Ca2+ channel inhibitor, suppressed the Nod factor-dependent cytosolic [Ca2+] elevation. These results indicate that transient Ca2+ influx from extracellular fluid is one of the earliest responses of soybean cells to NodBj-V (C18:1, MeFuc) in a host-specific manner.

  2. Relationships of bradyrhizobia from the legumes Apios americana and Desmodium glutinosum.

    Science.gov (United States)

    Parker, M A

    1999-11-01

    Multilocus enzyme electrophoresis, partial 23S rRNA sequences, and nearly full-length 16S rRNA sequences all indicated high genetic similarity among root-nodule bacteria associated with Apios americana, Desmodium glutinosum, and Amphicarpaea bracteata, three common herbaceous legumes whose native geographic ranges in eastern North America overlap extensively. A total of 19 distinct multilocus genotypes (electrophoretic types [ETs]) were found among the 35 A. americana and 33 D. glutinosum isolates analyzed. Twelve of these ETs (representing 78% of all isolates) were either identical to ETs previously observed in A. bracteata populations, or differed at only one locus. Within both 23S and 16S rRNA genes, several isolates from A. americana and D. glutinosum were either identical to A. bracteata isolates or showed only single nucleotide differences. Growth rates and nitrogenase activities of A. bracteata plants inoculated with isolates from D. glutinosum were equivalent to levels found with native A. bracteata bacterial isolates, but none of the three A. americana isolates tested had high symbiotic effectiveness on A. bracteata. Phylogenetic analysis of both 23S and 16S rRNA sequences indicated that both A. americana and D. glutinosum harbored rare bacterial genotypes similar to Bradyrhizobium japonicum USDA 110. However, the predominant root nodule bacteria on both legumes were closely related to Bradyrhizobium elkanii.

  3. 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.

  4. 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.

  5. Biodiversity of amoebae and amoeba-resisting bacteria in a hospital water network.

    Science.gov (United States)

    Thomas, Vincent; Herrera-Rimann, Katia; Blanc, Dominique S; Greub, Gilbert

    2006-04-01

    Free-living amoebae (FLA) are ubiquitous organisms that have been isolated from various domestic water systems, such as cooling towers and hospital water networks. In addition to their own pathogenicity, FLA can also act as Trojan horses and be naturally infected with amoeba-resisting bacteria (ARB) that may be involved in human infections, such as pneumonia. We investigated the biodiversity of bacteria and their amoebal hosts in a hospital water network. Using amoebal enrichment on nonnutrient agar, we isolated 15 protist strains from 200 (7.5%) samples. One thermotolerant Hartmannella vermiformis isolate harbored both Legionella pneumophila and Bradyrhizobium japonicum. By using amoebal coculture with axenic Acanthamoeba castellanii as the cellular background, we recovered at least one ARB from 45.5% of the samples. Four new ARB isolates were recovered by culture, and one of these isolates was widely present in the water network. Alphaproteobacteria (such as Rhodoplanes, Methylobacterium, Bradyrhizobium, Afipia, and Bosea) were recovered from 30.5% of the samples, mycobacteria (Mycobacterium gordonae, Mycobacterium kansasii, and Mycobacterium xenopi) were recovered from 20.5% of the samples, and Gammaproteobacteria (Legionella) were recovered from 5.5% of the samples. No Chlamydia or Chlamydia-like organisms were recovered by amoebal coculture or detected by PCR. The observed strong association between the presence of amoebae and the presence of Legionella (P amoebae when water control measures are designed.

  6. 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.

  7. Diversity of rhizobia nodulating wild shrubs of Sicily and some neighbouring islands.

    Science.gov (United States)

    Cardinale, Massimiliano; Lanza, Angela; Bonnì, Maria Laura; Marsala, Salvatore; Puglia, Anna Maria; Quatrini, Paola

    2008-10-01

    Legume shrubs have great potential for rehabilitation of semi-arid degraded soils in Mediterranean ecosystems as they establish mutualistic symbiosis with N-fixing rhizobia. Eighty-eight symbiotic rhizobia were isolated from seven wild legume shrubs native of Sicily (Southern Italy) and grouped in operational taxonomic units (OTU) by analysis of the ribosomal internal transcribed spacer (ITS) polymorphism. Partial sequencing of 16S rRNA gene of representative isolates of each OTU revealed that most Genisteae symbionts are related to Bradyrhizobium canariense, B. japonicum and B. elkanii. Teline monspessulana was the only Genistea nodulated by Mesorhizobium strains, and Anagyris foetida (Thermopsideae) was promiscuosly nodulated by Rhizobium, Mesorhizobium, Agrobacterium and Bradyrhizobium strains. Analysis of the nodulation gene nodA assigned most Mediterranean Genisteae bradyrhizobia to clade II but also to clades IV, I and III, which included, so far, sequences of (sub)tropical and Australian isolates. The high diversity and low host specificity observed in most wild legumes isolates suggest that preferential associations may establish in the field depending on differences in the benefits conferred to the host and on competition ability. Once identified, these beneficial symbiosis can be exploited for rehabilitation of arid, low productive and human-impacted soils of the Mediterranean countries.

  8. 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.

  9. 日本血吸虫重组质粒pGEX-Sj14-3-3-Sj32的构建及其在大肠埃希菌BL21(DE3)中的表达%Construction and expression of a recombinant plasmid pGEX-Sj14-3-3-Sj32 of Schistosoma japonicum in Escherichia coli BL21(DE3)

    Institute of Scientific and Technical Information of China (English)

    覃婷; 李文桂; 谭建蓉

    2015-01-01

    目的 构建日本血吸虫重组质粒pGEX-Sj14-3-3-Sj32,探讨重组质粒在大肠埃希菌BL21 (DE3)中的表达.方法 以重庆医科大学附属第一医院传染病寄生虫病研究所保存的质粒pGEX-Sj14-3-3和pET28α-Sj32为模板,通过PCR扩增Sj14-3-3和Sj32抗原编码基因,然后采用基因拼接法(gene SOEing)剪接Sj14-3-3和Sj32基因,得到Sj14-3-3-Sj32融合基因,定向克隆入穿梭载体pGEX-1 λT,构建重组质粒pGEX-Sj14-3-3-Sj32,并采用双酶切法进行验证.将重组质粒转化大肠埃希菌BL21 (DE3),经异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达后,用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹(Western blot)法对表达产物进行分析鉴定.结果 基因拼接法得到约1 750 bp的Sj14-3-3-Sj32融合基因;双酶切证实Sj14-3-3-Sj32融合基因成功插入pGEX-1λT载体中;SDS-PAGE分析显示,表达产物为相对分子质量约为73×103的重组蛋白;Western blot显示,重组蛋白可被日本血吸虫感染的兔血清识别.结论 成功构建日本血吸虫重组质粒pGEX-Sj 14-3-3-Sj32,该重组质粒在大肠埃希菌BL21(DE3)中得到了高效融合表达,且表达的融合蛋白具有特异的抗原性.%Objective To construct and express a recombinant plasmid pGEX-Sj 14-3-3-Sj32 of Schistosoma japonicum in Escherichia coli (E.Coli) BL21 (DE3).Methods Sj14-3-3 and Sj32 antigen genes were amplified by PCR from template of plasmids pGEX-Sj14-3-3 and pET28α-Sj32 which were extracted from recombinant bacteria BL21 (pET28α-Sj32) and BL21 (pGEX-Sj14-3-3) stored in Institute of Infectious and Parasitic Disease of the First Affiliated Hospital of Chongqing Medical University.Sj14-3-3-Sj32 fusion gene obtained with gene SOEing was cloned into the vector pGEX-1λT to construct pGEX-Sj14-3-3-Sj32 which was identified by double digestion.The recombinant plasmid pGEX-Sj 14-3-3-Sj32 was transformed into E.Coli BL21 (DE3).The recombinant strains were induced by

  10. Distinct changes in soybean xylem sap proteome in response to pathogenic and symbiotic microbe interactions

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    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.

  11. Lotus corniculatus nodulation specificity is changed by the presence of a soybean lectin gene

    Science.gov (United States)

    van Rhijn P; Goldberg; Hirsch

    1998-08-01

    Plant lectins have been implicated as playing an important role in mediating recognition and specificity in the Rhizobium-legume nitrogen-fixing symbiosis. To test this hypothesis, we introduced the soybean lectin gene Le1 either behind its own promoter or behind the cauliflower mosaic virus 35S promoter into Lotus corniculatus, which is nodulated by R. loti. We found that nodulelike outgrowths developed on transgenic L. corniculatus plant roots in response to Bradyrhizobium japonicum, which nodulates soybean and not Lotus spp. Soybean lectin was properly targeted to L. corniculatus root hairs, and although infection threads formed, they aborted in epidermal or hypodermal cells. Mutation of the lectin sugar binding site abolished infection thread formation and nodulation. Incubation of bradyrhizobia in the nodulation (nod) gene-inducing flavonoid genistein increased the number of nodulelike outgrowths on transgenic L. corniculatus roots. Studies of bacterial mutants, however, suggest that a component of the exopolysaccharide surface of B. japonicum, rather than Nod factor, is required for extension of host range to the transgenic L. corniculatus plants.

  12. 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

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    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.

  13. 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.

  14. 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.

  15. 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...

  16. 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.

  17. Evaluation of the reaction oof interspecific hybrids of common bean and tepary bean to Bradyrhizobium y Rhizobium

    Science.gov (United States)

    Interspecific hybrids between common bean, Phaseolus vulgaris L., and tepary bean, Phaseolus acutifolius A. Gray, have the potential to increase bean production in regions where rainfall is limited. In 2014, an experiment was initiated using a split-plot design. The treatments included inoculation, ...

  18. 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

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

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    Francisco Pérez-Montaño

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

  20. EFEITOS DA INOCULAÇÃO COM BACTÉRIAS DIAZOTRÓFICAS E DA ADUBAÇÃO NITROGENADA NO CRESCIMENTO E NA QUALIDADE DE MUDAS DE Inga laurina (SW. Willd. (Fabaceae1

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    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.

  1. TRATAMENTO DE SEMENTES DE ARROZ, TRIGO, FEIJÃO E SOJA COM UM PRODUTO FORMULADO À BASE DE CÉLULAS E DE METABÓLITOS DE Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    E LAZZARETTI

    1997-01-01

    Full Text Available Um produto constituido de pó-molhável (PBBS formulado à base de células (60g e de metabólitos (60g de Bacillus subtilis, argila (480g, espalhante (7,92g e água (2400ml, moído e seco, mostrou-se semelhante aos tratamentos com fungicidas recomendados para o controle de Rhizoctonia solani, Aspergillus sp. e Sclerotinia sclerotiorum em sementes de feijão; Pyricularia oryzae e Rhinchosporium sativum em sementes de arroz; e Cercospora kikuchii, Phomopsis phaseoli e Fusarium spp. em sementes de soja. Para Dreschlera oryzae em arroz e Bipolaris sorokiniana, P. oryzae e Alternaria tenuis em sementes de trigo, o tratamento com o PBBS, embora não tenha se igualado ao tratamento com o fungicida padrão, diferiu estatisticamente do tratamento testemunha. A nodulação das raízes de feijão e soja por bactérias simbióticas, fixadoras de nitrogênio, não foi influenciada quando o PBBS foi aplicado simultaneamente ao inoculante contendo Rhyzobium sp. e Bradyrhizobium japonicum, respectivamente. O PBBS também não afetou a emergência das plântulas das culturas testadas.A biological fungicide product containing B. subtilis cells (60 g and metabolites (60g was transformed into a wettable powder formulated with, clay (480 g, surfactant (7,92 g and water (2400 ml.The product showed to be similar to the conventional fungicides used in the control of Rhizoctonia solani, Aspergillus sp. and Sclerotinia sclerotiorum in bean seeds. The control of Pyricularia oryzae and Rhinchosporium sativum in rice seeds and Cercospora kikuchii, Phomopsis phaseoli and Fusarium spp. in soybean seeds was also as efficient as the comercial fungicides. For Dreschlera oryzae in rice seeds and Bipolaris sorokiniana, P. oryzae and Alternaria tenuis in wheat seeds, the treatment with the product, althought not as efficient as the chemical treatment, was statistically distinct from the control. The root nodulation of bean and soybean by nitrogen fixing bacteria, was not affected

  2. 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

  3. Rhizobial position as a main determinant in the problem of competition for nodulation in soybean.

    Science.gov (United States)

    López-García, Silvina L; Vázquez, Tirso E E; Favelukes, Gabriel; Lodeiro, Aníbal R

    2002-04-01

    Selected Bradyrhizobium japonicum strains inoculated on soybean seeds often fail to occupy a significant proportion of nodules when a competitor rhizobial population is established in the soil. This competition problem could result from a genetic/ physiological advantage of the adapted soil population over the introduced inoculant or from a positional advantage, as the soil population already occupies the soil profile where the roots will penetrate, whereas the inoculant remains concentrated around the seeds. Here, we have assessed the contribution of these factors with a laboratory model in which a rhizobial population is established in sterile vermiculite. We observed that the wild-type strain B. japonicum LP 3004 was able to grow in pots with N-free plant nutrient solution-watered vermiculite for six or seven generations with a duplication rate of at least 0.7 day(-1). In addition, the rhizobial population persisted for 3 months with 10(6)-10(7) colony-forming units ml(-1) of the vermiculite-retained solution. N-starved, young rhizobial cultures are more efficient in performing several steps along their early association with soybean roots. However, N starvation during growth of rhizobia used for seed inoculation did not enhance their competitiveness against a 1 month vermiculite-established rhizobial population, which occupied more than 72% of the nodules. When a similarly established rhizobial population was recovered from the vermiculite and homogeneously suspended in plant nutrient solution, these cells were significantly less competitive (29% of nodules occupied) than rhizobia obtained from a fresh, logarithmic culture in a N-poor minimal medium, thus indicating that cell position rather than intrinsic competitiveness was the determinant for nodule occupation.

  4. Solubilisation of inorganic phosphates by inoculant strains from tropical legumes

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    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.

  5. Environ: E00579 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00579 Processed aconitum japonicum tuberous root Crude drug Aconitum japonicum, Ac...t with caustic lime (dried) Crude drugs [BR:br08305] Dicot plants: others Ranunculaceae (buttercup family) E00579 Processed aconitum japonicum tuberous root ...

  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. 根瘤菌基因组内简单重复序列的分析%Analysis of Simple Sequence Repeats in Rhizobium Genomes

    Institute of Scientific and Technical Information of China (English)

    高亚梅; 韩毅强; 汤辉; 孙东梅; 王彦杰; 王伟东

    2008-01-01

    [目的]分析根瘤菌基因组中的简单重复序列(simple sequence repeats,SSRs),为其在根瘤菌遗传多样性研究中的应用提供有益的信息.[方法]利用公共的微生物串联重复序列数据库资源,对已测序的3种根瘤菌基因组中SSRs的结构类型,分布,丰度等进行系统的比较分析.[结果]大豆慢生根瘤菌(Bradyrhizobium japonicum)、百脉根根瘤菌(blesorhizobium loti)和苜蓿中华根瘤菌(Sinorhizobium meliloti)基因组中的SSRs分别为1 410个、859个和638个,3种根瘤菌基因组中长重复的四、五、六核苷酸基序更为丰富,变异性更高.数目最少的为单碱基重复.[结论]3种根瘤菌的SSR在结构类型和分布规律上均具有一定的相似性.

  8. Denitrification in Sinorhizobium meliloti.

    Science.gov (United States)

    Torres, María J; Rubia, María I; Bedmar, Eulogio J; Delgado, María J

    2011-12-01

    Denitrification is the complete reduction of nitrate or nitrite to N2, via the intermediates nitric oxide (NO) and nitrous oxide (N2O), and is coupled to energy conservation and growth under O2-limiting conditions. In Bradyrhizobium japonicum, this process occurs through the action of the napEDABC, nirK, norCBQD and nosRZDFYLX gene products. DNA sequences showing homology with nap, nirK, nor and nos genes have been found in the genome of the symbiotic plasmid pSymA of Sinorhizobium meliloti strain 1021. Whole-genome transcriptomic analyses have demonstrated that S. meliloti denitrification genes are induced under micro-oxic conditions. Furthermore, S. meliloti has also been shown to possess denitrifying activities in both free-living and symbiotic forms. Despite possessing and expressing the complete set of denitrification genes, S. meliloti is considered a partial denitrifier since it does not grow under anaerobic conditions with nitrate or nitrite as terminal electron acceptors. In the present paper, we show that, under micro-oxic conditions, S. meliloti is able to grow by using nitrate or nitrite as respiratory substrates, which indicates that, in contrast with anaerobic denitrifiers, O2 is necessary for denitrification by S. meliloti. Current knowledge of the regulation of S. meliloti denitrification genes is also included.

  9. 苜蓿中华根瘤菌matB基因的克隆及其功能的研究%Cloning and Functional Analysis of matB Gene in Sinorhizobium meliloti

    Institute of Scientific and Technical Information of China (English)

    王永宝; 孙杰; 刘君; 陈爱民; 王彦章

    2009-01-01

    通过同源性分析,发现苜蓿中华根瘤菌(Sinorhizobium meliloti)菌株Rm1021中matB基因与三叶草生物型豌豆根瘤菌(Rhizobium leguminosarum by.trifolii)和慢生型大豆根瘤菌(Bradyrhizobium japonicum USDA110)中编码丙二酸单酰辅酶A合成酶(malonyl-CoA synthetase)基因在氨基酸水平上分别达到了75%和67%一致性,具有高度同源性.因此,从Rm1021中克隆出matB基因,并在大肠埃希菌(Escherichia coli)中进行体外诱导表达和纯化.纯化的MalB蛋白具有丙二酸单酰辅酶A合成酶的活性.测定的Km值是710 μmol,Vmax是0.209 μmol/min/mg.

  10. HmuS and HmuQ of Ensifer/Sinorhizobium meliloti degrade heme in vitro and participate in heme metabolism in vivo.

    Science.gov (United States)

    Amarelle, Vanesa; Rosconi, Federico; Lázaro-Martínez, Juan Manuel; Buldain, Graciela; Noya, Francisco; O'Brian, Mark R; Fabiano, Elena

    2016-04-01

    Ensifer meliloti is a nitrogen-fixing symbiont of the alfalfa legume able to use heme as an iron source. The transport mechanism involved in heme acquisition in E. meliloti has been identified and characterized, but the fate of heme once inside the cell is not known. In silico analysis of E. meliloti 1021 genome revealed no canonical heme oxygenases although two genes encoding putative heme degrading enzymes, smc01518 and hmuS, were identified. SMc01518 is similar to HmuQ of Bradyrhizobium japonicum, which is weakly homologous to the Staphylococcus aureus IsdG heme-degrading monooxygenase, whereas HmuS is homolog to Pseudomonas aeruginosa PhuS, a protein reported as a heme chaperone and as a heme degrading enzyme. Recombinant HmuQ and HmuS were able to bind hemin with a 1:1 stoichiometry and displayed a Kd value of 5 and 4 µM, respectively. HmuS degrades heme in vitro to the biliverdin isomers IX-β and IX-δ in an equimolar ratio. The HmuQ recombinant protein degrades heme to biliverdin IX-δ only. Additionally, in this work we demonstrate that humS and hmuQ gene expression is regulated by iron and heme in a RirA dependent manner and that both proteins are involved in heme metabolism in E. meliloti in vivo.

  11. Dual control of Sinorhizobium meliloti RpoE2 sigma factor activity by two PhyR-type two-component response regulators.

    Science.gov (United States)

    Bastiat, Bénédicte; Sauviac, Laurent; Bruand, Claude

    2010-04-01

    RpoE2 is an extracytoplasmic function (ECF) sigma factor involved in the general stress response of Sinorhizobium meliloti, the nitrogen-fixing symbiont of the legume plant alfalfa. RpoE2 orthologues are widely found among alphaproteobacteria, where they play various roles in stress resistance and/or host colonization. In this paper, we report a genetic and biochemical investigation of the mechanisms of signal transduction leading to S. meliloti RpoE2 activation in response to stress. We showed that RpoE2 activity is negatively controlled by two paralogous anti-sigma factors, RsiA1 (SMc01505) and RsiA2 (SMc04884), and that RpoE2 activation by stress requires two redundant paralogous PhyR-type response regulators, RsiB1 (SMc01504) and RsiB2 (SMc00794). RsiB1 and RsiB2 do not act at the level of rpoE2 transcription but instead interact with the anti-sigma factors, and we therefore propose that they act as anti-anti-sigma factors to relieve RpoE2 inhibition in response to stress. This model closely resembles a recently proposed model of activation of RpoE2-like sigma factors in Methylobacterium extorquens and Bradyrhizobium japonicum, but the existence of two pairs of anti- and anti-anti-sigma factors in S. meliloti adds an unexpected level of complexity, which may allow the regulatory system to integrate multiple stimuli.

  12. Nodule Formation and Development in Soybeans (Glycine max L.) in Response to Phosphorus Supply in Solution Culture

    Institute of Scientific and Technical Information of China (English)

    MIAO Shu-Jie; QIAO Yun-Fa; HAN Xiao-Zeng; M. AN

    2007-01-01

    Phosphorus (P) is necessary for growth and nitrogen fixation, and thus its deficiency is a major factor limiting legume production in most agricultural soils. The effect of phosphorus supply on nodule development and its role in soybeans (Glycine max L.) was studied in a nutrient solution. Plants were inoculated with Bradyrhizobium japonicum and grown for 35 days in a glasshouse at a day and night temperature of 25 ℃ and 15 ℃, respectively. Although increasing P supply increased the concentrations of P and N in the shoots and roots, the external P supply did not significantly affect the P concentration in the nodules, and the N fixed per unit nodule biomass decreased with increasing P supply. The nitrogen content in the shoots correlated well with the P content (r = 0.92**). At an inoculation level of 102 cells mL-1, the P supply did not affect the number of nodules; however, at inoculation levels of 103.5 and 105 cells mL-1, increasing P supply increased both the number and size of nodules. Irrespective of the inoculation level, increasing P supply increased the nodule biomass relative to the biomass of the host plant. It is suggested that the P deficiency specifically inhibited the nodule development and thereby the total N2 fixation.

  13. The ccoNOQP gene cluster codes for a cb-type cytochrome oxidase that functions in aerobic respiration of Rhodobacter capsulatus.

    Science.gov (United States)

    Thöny-Meyer, L; Beck, C; Preisig, O; Hennecke, H

    1994-11-01

    The genes for a new type of a haem-copper cytochrome oxidase were cloned from Rhodobacter capsulatus strain 37b4, using the Bradyrhizobium japonicum fixNOQP gene region as a hybridizing probe. Four genes, probably organized in an operon (ccoNOQP), were identified; their products share extensive amino acid sequence similarity with the FixN, O, Q and P proteins that have recently been shown to be the subunits of a cb-type oxidase. CcoN is a b-type cytochrome, CcoO and CcoP are membrane-bound mono- and dihaem c-type cytochromes and CcoQ is a small membrane protein of unknown function. Genes for a similar oxidase are also present in other non-rhizobial bacterial species such as Azotobacter vinelandii, Agrobacterium tumefaciens and Pseudomonas aeruginosa, as revealed by polymerase chain reaction analysis. A ccoN mutant was constructed whose phenotype, in combination with the structural information on the gene products, provides evidence that the CcoNOQP oxidase is a cytochrome c oxidase of the cb type, which supports aerobic respiration in R. capsulatus and which is probably identical to the cbb3-type oxidase that was recently purified from a different strain of the same species. Mutant analysis also showed that this oxidase has no influence on photosynthetic growth and nitrogen-fixation activity.

  14. Phytotoxicity attenuation in Vigna radiata under heavy metal stress at the presence of biochar and N fixing bacteria.

    Science.gov (United States)

    Seneviratne, Mihiri; Weerasundara, Lakshika; Ok, Yong Sik; Rinklebe, Jörg; Vithanage, Meththika

    2017-01-15

    This study assesses the effect of N-fixing bacteria and biochar synergism on plant growth and development of Vigna mungo under heavy metal stress (HM). Heavy metal stress is a worldwide problem, which causes critical effects on plant life due to oxidative stress. Application of biochar is a recent biological remediation technique, which often leads to an immobilization of heavy metals in soil. . Synergism of bacteria and biochar is a novel aspect to enhance plant growth under heavy metal stress. Woody biochar a byproduct of a dendro power industry was added as 1, 2.5 and 5% amounts combination with Bradyrhizobium japonicum, where mung seedlings were planted in serpentine soil rich in Ni, Mn, Cr and Co. Pot experiments were conducted for 12 weeks. The plant height, heavy metal uptake by plants, soil bioavailable heavy metal contents, soil N and P and microbial biomass carbon (MBC) were measured. The plant growth was enhanced with biochar amendment but a retardation was observed with high biochar application (5%). The soil N and P increased with the increase of biochar addition percentage while soil MBC showed reductions at 5% biochar amendment. Both soil bioavailable fractions of HM and up take of HMs by plants were gradually reduced with increase in biochar content. Based on the results, 2.5% biochar synergism with bacteria was the best for plant growth and soil nutrition status. Despite the synergism, available N was negatively correlated with the decrease of bioavailable metal percentage in soil whereas it was conversely for P.

  15. 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

  16. X-ray microanalytical studies of mineral elements in the tripartite symbiosis between lima bean, N2-fixing bacteria and mycorrhizal fungi.

    Science.gov (United States)

    Rodak, Bruna Wurr; Freitas, Douglas Siqueira; Bamberg, Soraya Marx; Carneiro, Marco Aurélio Carbone; Guilherme, Luiz Roberto Guimarães

    2017-01-01

    The symbiosis between legumes, arbuscular mycorrhizal (AM) fungi, and N2-fixing bacteria (NFB) provides mutual nutritional gains. However, assessing the nutritional status of the microorganisms is a difficult task. A methodology that could assess this status, in situ, could assist managing these organisms in agriculture. This study used X-ray microanalyses to quantify and locate mineral elements in structures formed in a tripartite symbiosis. Lima bean (Phaseolus lunatus L. Walp) was cultivated in pots under greenhouse conditions, to which we have added AM fungal isolates (Glomus macrocarpum and Acaulospora colombiana) and NFB (Bradyrhizobium japonicum) inocula. Uninoculated control plants were also included. Symbionts were evaluated at the onset of flowering. Quantification of the mineral elements in the symbiotic components was performed using energy dispersive X-ray spectroscopy (EDX) and a scanning electron microscopy (SEM) was used to identify structures. EDX analysis detected 13 elements with the most abundant being N, Ca, and Se, occurring in all tissues, Fe in roots, Ni and Al in epidermis and P and Mo in nodules. Elemental quantification in fungal structures was not possible. The distribution of elements was related to their symbiotic function. X-ray microanalysis can be efficiently applied for nutritional diagnosis in tripartite symbiosis.

  17. 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.

  18. 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.

  19. Functional conservation of the capacity for ent-kaurene biosynthesis and an associated operon in certain rhizobia.

    Science.gov (United States)

    Hershey, David M; Lu, Xuan; Zi, Jiachen; Peters, Reuben J

    2014-01-01

    Bacterial interactions with plants are accompanied by complex signal exchange processes. Previously, the nitrogen-fixing symbiotic (rhizo)bacterium Bradyrhizobium japonicum was found to carry adjacent genes encoding two sequentially acting diterpene cyclases that together transform geranylgeranyl diphosphate to ent-kaurene, the olefin precursor to the gibberellin plant hormones. Species from the three other major genera of rhizobia were found to have homologous terpene synthase genes. Cloning and functional characterization of a representative set of these enzymes confirmed the capacity of each genus to produce ent-kaurene. Moreover, comparison of their genomic context revealed that these diterpene synthases are found in a conserved operon which includes an adjacent isoprenyl diphosphate synthase, shown here to produce the geranylgeranyl diphosphate precursor, providing a critical link to central metabolism. In addition, the rest of the operon consists of enzymatic genes that presumably lead to a more elaborated diterpenoid, although the production of gibberellins was not observed. Nevertheless, it has previously been shown that the operon is selectively expressed during nodulation, and the scattered distribution of the operon via independent horizontal gene transfer within the symbiotic plasmid or genomic island shown here suggests that such diterpenoid production may modulate the interaction of these particular symbionts with their host plants.

  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. Plant dependence on rhizobia for nitrogen influences induced plant defenses and herbivore performance.

    Science.gov (United States)

    Dean, Jennifer M; Mescher, Mark C; De Moraes, Consuelo M

    2014-01-21

    Symbiotic rhizobia induce many changes in legumes that could affect aboveground interactions with herbivores. We explored how changing the intensity of Bradyrhizobium japonicum, as modulated by soil nitrogen (N) levels, influenced the interaction between soybean (Glycine max) and herbivores of different feeding guilds. When we employed a range of fertilizer applications to manipulate soil N, plants primarily dependent on rhizobia for N exhibited increased root nodulation and higher levels of foliar ureides than plants given N fertilizer; yet all treatments maintained similar total N levels. Soybean podworm (Helicoverpa zea) larvae grew best on plants with the highest levels of rhizobia but, somewhat surprisingly, preferred to feed on high-N-fertilized plants when given a choice. Induction of the defense signaling compound jasmonic acid (JA) by H. zea feeding damage was highest in plants primarily dependent on rhizobia. Differences in rhizobial dependency on soybean did not appear to affect interactions with the phloem-feeding soybean aphid (Aphis glycines). Overall, our results suggest that rhizobia association can affect plant nutritional quality and the induction of defense signaling pathways and that these effects may influence herbivore feeding preferences and performance-though such effects may vary considerably for different classes of herbivores.

  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. Indole-3-acetic acid biosynthesis is deficient in Gluconacetobacter diazotrophicus strains with mutations in cytochrome c biogenesis genes.

    Science.gov (United States)

    Lee, Sunhee; Flores-Encarnación, M; Contreras-Zentella, M; Garcia-Flores, L; Escamilla, J E; Kennedy, Christina

    2004-08-01

    Gluconacetobacter diazotrophicus is an endophyte of sugarcane frequently found in plants grown in agricultural areas where nitrogen fertilizer input is low. Recent results from this laboratory, using mutant strains of G. diazotrophicus unable to fix nitrogen, suggested that there are two beneficial effects of G. diazotrophicus on sugarcane growth: one dependent and one not dependent on nitrogen fixation. A plant growth-promoting substance, such as indole-3-acetic acid (IAA), known to be produced by G. diazotrophicus, could be a nitrogen fixation-independent factor. One strain, MAd10, isolated by screening a library of Tn5 mutants, released only approximately 6% of the amount of IAA excreted by the parent strain in liquid culture. The mutation causing the IAA(-) phenotype was not linked to Tn5. A pLAFR3 cosmid clone that complemented the IAA deficiency was isolated. Sequence analysis of a complementing subclone indicated the presence of genes involved in cytochrome c biogenesis (ccm, for cytochrome c maturation). The G. diazotrophicus ccm operon was sequenced; the individual ccm gene products were 37 to 52% identical to ccm gene products of Escherichia coli and equivalent cyc genes of Bradyrhizobium japonicum. Although several ccm mutant phenotypes have been described in the literature, there are no reports of ccm gene products being involved in IAA production. Spectral analysis, heme-associated peroxidase activities, and respiratory activities of the cell membranes revealed that the ccm genes of G. diazotrophicus are involved in cytochrome c biogenesis.

  4. nodSU, two new nod genes of the broad host range Rhizobium strain NGR234 encode host-specific nodulation of the tropical tree Leucaena leucocephala.

    Science.gov (United States)

    Lewin, A; Cervantes, E; Chee-Hoong, W; Broughton, W J

    1990-01-01

    Rhizobium species strain NGR234 nodulates at least 35 diverse genera of legumes as well as the nonlegume Parasponia andersonii. Most nodulation genes are located on the 500-kilobase pair symbiotic plasmid, pNGR234a. Previously, three plasmid-borne host range determinants (HsnI, HsnII, and HsnIII) were identified by their ability to extend the nodulation capacity of heterologous rhizobia to include Vigna unguiculata. In this study, we show that HsnII contains two new nod-box linked hsn genes, nodS and nodU.nodS controls nodulation of the tropical tree Leucaena leucocephala, while the nodSU genes regulate nodulation of the pasture legume Desmodium intortum and the grain legume V. unguiculata. Regulation of the nod-box upstream of nodSU by the flavonoid naringenin was shown using a fusion with a promoterless lacZ gene. Determination of the nucleotide sequence of the nodS gene did not reveal homology with any gene in the EMBL library, although Bradyrhizobium japonicum USDA110 contains both nodS and nodU (M. Göttfert, S. Hitz, and H. Hennecke, Molecular Plant-Microbe Interactions 3:308-316, 1990). We suggest that broad host range in NGR234 is controlled in part by a nodD gene which interacts with a wide range of flavonoids, and in part by host-specific nod genes such as nodS.

  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. Exposure versus Susceptibility as Alternative Bases for New Approaches to Surveillance for Schistosoma japonicum in Low Transmission Environments.

    Directory of Open Access Journals (Sweden)

    Shuo Wang

    2016-03-01

    Full Text Available Currently, schistosomiasis in China provides an excellent example of many of the challenges of moving from low transmission to the elimination of transmission for infectious diseases generally. In response to the surveillance dimension of these challenges, we here explore two strategic approaches to inform priorities for the development of improved methods addressed specifically to schistosomiasis in the low transmission environment. We utilize an individually-based model and the exposure data used earlier to explore surveillance strategies, one focused on exposure assessment and the second on our estimates of variability in individual susceptibility in the practical context of the current situation in China and the theoretical context of the behavior of transmission dynamics near the zero state. Our findings suggest that individual susceptibility is the major single determinant of infection intensity in both the low and medium risk environments. We conclude that there is considerable motivation to search for a biomarker of susceptibility to infection in humans, but that there would also be value in a method for monitoring surface waters for the free-swimming forms of the parasite in endemic or formerly endemic environments as an early warning of infection risk.

  7. 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.

  8. NCBI nr-aa BLAST: CBRC-TTRU-01-1326 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-1326 emb|CAX75571.1| Transmembrane protein 118 [Schistosoma japonicum]... emb|CAX75572.1| Transmembrane protein 118 [Schistosoma japonicum] emb|CAX75574.1| Transmembrane protein 118 [Schistosoma... japonicum] emb|CAX75575.1| Transmembrane protein 118 [Schistosoma japonicum] CAX75571.1 0.36 27% ...

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

    Lifescience Database Archive (English)

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

  10. NCBI nr-aa BLAST: CBRC-MMUR-01-0199 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-0199 emb|CAX76110.1| putative tubulin, beta, 2 [Schistosoma japonicum]... emb|CAX76111.1| putative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76112.1| putative tubulin, beta, 2 [Schistosoma... japonicum] emb|CAX76113.1| putative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76114.1| p...utative tubulin, beta, 2 [Schistosoma japonicum] emb|CAX76115.1| putative tubulin, beta, 2 [Schistosoma... japonicum] emb|CAX76116.1| putative tubulin, beta, 2 [Schistosoma japonicum] CAX76110.1 2e-31 82% ...

  11. 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

  12. 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

  13. Conservation of gene order and content in the circular chromosomes of 'Candidatus Liberibacter asiaticus' and other Rhizobiales.

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    L David Kuykendall

    Full Text Available 'Ca. Liberibacter asiaticus,' an insect-vectored, obligate intracellular bacterium associated with citrus-greening disease, also called "HLB," is a member of the Rhizobiales along with nitrogen-fixing microsymbionts Sinorhizobium meliloti and Bradyrhizobium japonicum, plant pathogen Agrobacterium tumefaciens and facultative intracellular mammalian pathogen Bartonella henselae. Comparative analyses of their circular chromosomes identified 514 orthologous genes shared among all five species. Shared among all five species are 50 identical blocks of microsyntenous orthologous genes (MOGs, containing a total of 283 genes. While retaining highly conserved genomic blocks of microsynteny, divergent evolution, horizontal gene transfer and niche specialization have disrupted macrosynteny among the five circular chromosomes compared. Highly conserved microsyntenous gene clusters help define the Rhizobiales, an order previously defined by 16S RNA gene similarity and herein represented by the three families: Bartonellaceae, Bradyrhizobiaceae and Rhizobiaceae. Genes without orthologs in the other four species help define individual species. The circular chromosomes of each of the five Rhizobiales species examined had genes lacking orthologs in the other four species. For example, 63 proteins are encoded by genes of 'Ca. Liberibacter asiaticus' not shared with other members of the Rhizobiales. Of these 63 proteins, 17 have predicted functions related to DNA replication or RNA transcription, and some of these may have roles related to low genomic GC content. An additional 17 proteins have predicted functions relevant to cellular processes, particularly modifications of the cell surface. Seventeen unshared proteins have specific metabolic functions including a pathway to synthesize cholesterol encoded by a seven-gene operon. The remaining 12 proteins encoded by 'Ca. Liberibacter asiaticus' genes not shared with other Rhizobiales are of bacteriophage origin. 'Ca

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

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    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.

  15. Comparison of the 'Ca. Liberibacter asiaticus' genome adapted for an intracellular lifestyle with other members of the Rhizobiales.

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    John S Hartung

    Full Text Available An intracellular plant pathogen 'Candidatus Liberibacter asiaticus,' a member of the Rhizobiales, is related to Sinorhizobium meliloti, Bradyrhizobium japonicum, nitrogen fixing endosymbionts, Agrobacterium tumefaciens, a plant pathogen, and Bartonella henselae, an intracellular mammalian pathogen. Whole chromosome comparisons identified at least 50 clusters of conserved orthologous genes found on the chromosomes of all five metabolically diverse species. The intracellular pathogens 'Ca. Liberibacter asiaticus' and Bartonella henselae have genomes drastically reduced in gene content and size as well as a relatively low content of guanine and cytosine. Codon and amino acid preferences that emphasize low guanosine and cytosine usage are globally employed in these genomes, including within regions of microsynteny and within signature sequences of orthologous proteins. The length of orthologous proteins is generally conserved, but not their isoelectric points, consistent with extensive amino acid substitutions to accommodate selection for low GC content. The 'Ca. Liberibacter asiaticus' genome apparently has all of the genes required for DNA replication present in Sinorhizobium meliloti except it has only two, rather than three RNaseH genes. The gene set required for DNA repair has only one rather than ten DNA ligases found in Sinorhizobium meliloti, and the DNA PolI of 'Ca. Liberibacter asiaticus' lacks domains needed for excision repair. Thus the ability of 'Ca. Liberibacter asiaticus' to repair mutations in its genome may be impaired. Both 'Ca. Liberibacter asiaticus and Bartonella henselae lack enzymes needed for the metabolism of purines and pyrimidines, which must therefore be obtained from the host. The 'Ca. Liberibacter asiaticus' genome also has a greatly reduced set of sigma factors used to control transcription, and lacks sigma factors 24, 28 and 38. The 'Ca. Liberibacter asiaticus' genome has all of the hallmarks of a reduced genome of a

  16. Genome sequence of the chemolithoautotrophic nitrite-oxidizing bacterium Nitrobacter winogradskyi Nb-255

    Energy Technology Data Exchange (ETDEWEB)

    Hauser, Loren John [ORNL; Land, Miriam L [ORNL; Larimer, Frank W [ORNL; Arp, D J [Oregon State University; Hickey, W J [University of Wisconsin, Madison

    2006-03-01

    The alphaproteobacterium Nitrobacter winogradskyi (ATCC 25391) is a gram-negative facultative chemolithoautotroph capable of extracting energy from the oxidation of nitrite to nitrate. Sequencing and analysis of its genome revealed a single circular chromosome of 3,402,093 bp encoding 3,143 predicted proteins. There were extensive similarities to genes in two alphaproteobacteria, Bradyrhizobium japonicum USDA110 (1,300 genes) and Rhodopseudomonas palustris CGA009 CG (815 genes). Genes encoding pathways for known modes of chemolithotrophic and chemoorganotrophic growth were identified. Genes encoding multiple enzymes involved in anapleurotic reactions centered on C2 to C4 metabolism, including a glyoxylate bypass, were annotated. The inability of N. winogradskyi to grow on C6 molecules is consistent with the genome sequence, which lacks genes for complete Embden-Meyerhof and Entner-Doudoroff pathways, and active uptake of sugars. Two gene copies of the nitrite oxidoreductase, type I ribulose-1,5-bisphosphate carboxylase/oxygenase, cytochrome c oxidase, and gene homologs encoding an aerobic-type carbon monoxide dehydrogenase were present. Similarity of nitrite oxidoreductases to respiratory nitrate reductases was confirmed. Approximately 10% of the N. winogradskyi genome codes for genes involved in transport and secretion, including the presence of transporters for various organic-nitrogen molecules. The N. winogradskyi genome provides new insight into the phylogenetic identity and physiological capabilities of nitrite-oxidizing bacteria. The genome will serve as a model to study the cellular and molecular processes that control nitrite oxidation and its interaction with other nitrogen-cycling processes.

  17. 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.

  18. Functional implication of β-carotene hydroxylases in soybean nodulation.

    Science.gov (United States)

    Kim, Yun-Kyoung; Kim, Sunghan; Um, Ji-Hyun; Kim, Kyunga; Choi, Sun-Kang; Um, Byung-Hun; Kang, Suk-Woo; Kim, Jee-Woong; Takaichi, Shinichi; Song, Seok-Bo; Lee, Choon-Hwan; Kim, Ho-Seung; Kim, Ki Woo; Nam, Kyoung Hee; Lee, Suk-Ha; Kim, Yul-Ho; Park, Hyang-Mi; Ha, Sun-Hwa; Verma, Desh Pal S; Cheon, Choong-Ill

    2013-07-01

    Legume-Rhizobium spp. symbiosis requires signaling between the symbiotic partners and differential expression of plant genes during nodule development. Previously, we cloned a gene encoding a putative β-carotene hydroxylase (GmBCH1) from soybean (Glycine max) whose expression increased during nodulation with Bradyrhizobium japonicum. In this work, we extended our study to three GmBCHs to examine their possible role(s) in nodule development, as they were additionally identified as nodule specific, along with the completion of the soybean genome. In situ hybridization revealed the expression of three GmBCHs (GmBCH1, GmBCH2, and GmBCH3) in the infected cells of root nodules, and their enzymatic activities were confirmed by functional assays in Escherichia coli. Localization of GmBCHs by transfecting Arabidopsis (Arabidopsis thaliana) protoplasts with green fluorescent protein fusions and by electron microscopic immunogold detection in soybean nodules indicated that GmBCH2 and GmBCH3 were present in plastids, while GmBCH1 appeared to be cytosolic. RNA interference of the GmBCHs severely impaired nitrogen fixation as well as nodule development. Surprisingly, we failed to detect zeaxanthin, a product of GmBCH, or any other carotenoids in nodules. Therefore, we examined the possibility that most of the carotenoids in nodules are converted or cleaved to other compounds. We detected the expression of some carotenoid cleavage dioxygenases (GmCCDs) in wild-type nodules and also a reduced amount of zeaxanthin in GmCCD8-expressing E. coli, suggesting cleavage of the carotenoid. In view of these findings, we propose that carotenoids such as zeaxanthin synthesized in root nodules are cleaved by GmCCDs, and we discuss the possible roles of the carotenoid cleavage products in nodulation.

  19. 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.

  20. Effect of mycorrhizae, Thiobacillus and sulfur nutrition on the chemical composition of soybean [Glycine max (L.)] Merr. seed.

    Science.gov (United States)

    Mostafavian, S R; Pirdashti, H; Ramzanpour, M R; Andarkhor, A A; Shahsavari, A

    2008-03-15

    A field experiment carried out in a calcareous soil with a low available phosphorus to evaluate effectiveness of biofertilizers, mycorrhizae (Glomus intraradices) and Thiobacillus sp. inoculation individually or in combination on seed yield, oil, protein and some elements (P, Fe, Mn, Zn) concentration in two soybean [Glycine max (L.) Merr.] cultivars. The applied treatments were different fertilizers with 6 levels (including: NP (control, 12 kg N ha(-1) as urea, 46 kg P2O5 ha(-1) as triple super phosphate); NPK (NP + 75 kg K2O ha(-1) as potassium sulphate); NPKS [NPK+ S (100 kg S ha(-1))]; NPKST (NPKS + seed inoculation with Thiobacillus bacteria); NPKM (NPK + Seed inoculation with mycorrhizae fungi) and NPKSTM (NPKS + seed inoculation with Thiobacillus and mycorrhizae) and two cultivars (JK and 032). Before planting, soybean seeds were inoculated by Bradyrhizobium japonicum in all treatments. Results showed that combined inoculation of biofertilizers increased yield, however the highest yield was observed in treatment NPKST. Increasing oil content (percentage) was more pronounced in treatments NPKM, while most protein content (percentage) increasing was observed in NPKS and NPKM. Fe and Zn concentrations were unaffected significantly by fertilizer treatments, but NPKSTM showed significantly higher value of seed's Mn concentration compared to treatments NP and NPK. Although no significant difference was observed in terms ofP concentration of 032 line among fertilizer treatments, JK cultivar and NPKSTM caused a significant increasing in P concentration compared to NP, NPKS and NPKM. Present results suggested that applying biofertilizers i.e., mycorrhizae and Thiobacillus increased soybean yield compared to control (NP). Overall, this study demonstrated that soybean seed yield and its chemical composition could be affected by biofertilizer inoculation.

  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. Complete genome sequence of Nitrobacter hamburgensis X14 and comparative genomic analysis of species within the genus Nitrobacter.

    Energy Technology Data Exchange (ETDEWEB)

    Starkenburg, Shawn R [Oregon State University; Larimer, Frank W [ORNL; Stein, Lisa Y [University of California, Riverside; Klotz, Martin G [University of Louisville, Louisville; Chain, Patrick S. G. [Lawrence Livermore National Laboratory (LLNL); Sayavedra-Soto, LA [Oregon State University; Poret-Peterson, Amisha T. [University of Louisville, Louisville; Gentry, ME [University of Louisville, Louisville; Arp, D J [Oregon State University; Ward, Bess B. [Princeton University; Bottomley, Peter J [Oregon State University

    2008-05-01

    The alphaproteobacterium Nitrobacter hamburgensis X14 is a gram-negative facultative chemolithoautotroph that conserves energy from the oxidation of nitrite to nitrate. Sequencing and analysis of the Nitrobacter hamburgensis X14 genome revealed four replicons comprised of one chromosome (4.4 Mbp) and three plasmids (294, 188, and 121 kbp). Over 20% of the genome is composed of pseudogenes and paralogs. Whole-genome comparisons were conducted between N. hamburgensis and the finished and draft genome sequences of Nitrobacter winogradskyi and Nitrobacter sp. strain Nb-311A, respectively. Most of the plasmid-borne genes were unique to N. hamburgensis and encode a variety of functions (central metabolism, energy conservation, conjugation, and heavy metal resistance), yet approximately 21 kb of a approximately 28-kb "autotrophic" island on the largest plasmid was conserved in the chromosomes of Nitrobacter winogradskyi Nb-255 and Nitrobacter sp. strain Nb-311A. The N. hamburgensis chromosome also harbors many unique genes, including those for heme-copper oxidases, cytochrome b(561), and putative pathways for the catabolism of aromatic, organic, and one-carbon compounds, which help verify and extend its mixotrophic potential. A Nitrobacter "subcore" genome was also constructed by removing homologs found in strains of the closest evolutionary relatives, Bradyrhizobium japonicum and Rhodopseudomonas palustris. Among the Nitrobacter subcore inventory (116 genes), copies of genes or gene clusters for nitrite oxidoreductase (NXR), cytochromes associated with a dissimilatory nitrite reductase (NirK), PII-like regulators, and polysaccharide formation were identified. Many of the subcore genes have diverged significantly from, or have origins outside, the alphaproteobacterial lineage and may indicate some of the unique genetic requirements for nitrite oxidation in Nitrobacter.

  3. Characterization of Flavin-Containing Opine Dehydrogenase from Bacteria.

    Directory of Open Access Journals (Sweden)

    Seiya Watanabe

    Full Text Available Opines, in particular nopaline and octopine, are specific compounds found in crown gall tumor tissues induced by infections with Agrobacterium species, and are synthesized by well-studied NAD(PH-dependent dehydrogenases (synthases, which catalyze the reductive condensation of α-ketoglutarate or pyruvate with L-arginine. The corresponding genes are transferred into plant cells via a tumor-inducing (Ti plasmid. In addition to the reverse oxidative reaction(s, the genes noxB-noxA and ooxB-ooxA are considered to be involved in opine catabolism as (membrane-associated oxidases; however, their properties have not yet been elucidated in detail due to the difficulties associated with purification (and preservation. We herein successfully expressed Nox/Oox-like genes from Pseudomonas putida in P. putida cells. The purified protein consisted of different α-, β-, and γ-subunits encoded by the OdhA, OdhB, and OdhC genes, which were arranged in tandem on the chromosome (OdhB-C-A, and exhibited dehydrogenase (but not oxidase activity toward nopaline in the presence of artificial electron acceptors such as 2,6-dichloroindophenol. The enzyme contained FAD, FMN, and [2Fe-2S]-iron sulfur as prosthetic groups. On the other hand, the gene cluster from Bradyrhizobium japonicum consisted of OdhB1-C-A-B2, from which two proteins, OdhAB1C and OdhAB2C, appeared through the assembly of each β-subunit together with common α- and γ-subunits. A poor phylogenetic relationship was detected between OdhB1 and OdhB2 in spite of them both functioning as octopine dehydrogenases, which provided clear evidence for the acquisition of novel functions by "subunit-exchange". To the best of our knowledge, this is the first study to have examined flavin-containing opine dehydrogenase.

  4. 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.

  5. Effects of Salinity on Leaf Spectral Reflectance and Biochemical Parameters of Nitrogen Fixing Soybean Plants (Glycine max L.)

    Science.gov (United States)

    Krezhova, Dora D.; Kirova, Elisaveta B.; Yanev, Tony K.; Iliev, Ilko Ts.

    2010-01-01

    Measurements of physiology and hyperspectral leaf reflectance were used to detect salinity stress in nitrogen fixing soybean plants. Seedlings were inoculated with suspension of Bradyrhizobium japonicum strain 273. Salinity was performed at the stage of 2nd-4th trifoliate expanded leaves by adding of NaCl in the nutrient solution of Helrigel in concentrations 40 mM and 80 mM. A comparative analysis was performed between the changes in the biochemical parameters - stress markers (phenols, proline, malondialdehyde, thiol groups), chlorophyll a and b, hydrogen peroxide, and leaf spectral reflectance in the spectral range 450-850 nm. The spectral measurements were carried out by an USB2000 spectrometer. The reflectance data of the control and treated plants in the red, green, red-edge and the near infrared ranges of the spectrum were subjected to statistical analysis. Statistically significant differences were found through the Student's t-criterion at the two NaCl concentrations in all of the ranges examined with the exception of the near infrared range at 40 mM NaCl concentration. Similar results were obtained through linear discriminant analysis. The tents of the phenols, malondialdehyde and chlorophyll a and b were found to decrease at both salinity treatments. In the spectral data this effect is manifested by decrease of the reflectance values in the green and red ranges. The contents of proline, hydrogen peroxide and thiol groups rose with the NaCl concentration increase. At 80 mM NaCl concentration the values of these markers showed a considerable increase giving evidence that the soybean plants were stressed in comparison with the control. This finding is in agreement with the results from the spectral reflectance analysis.

  6. Symbiotic capability of calopo rhizobia from an agrisoil with different crops in Pernambuco

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    Altanys Silva Calheiros

    2013-08-01

    Full Text Available Biological nitrogen fixation by rhizobium-legume symbiosis represents one of the most important nitrogen sources for plants and depends strongly on the symbiotic efficiency of the rhizobium strain. This study evaluated the symbiotic capacity of rhizobial isolates from calopo (CALOPOGONIUM MUCUNOIDES taken from an agrisoil under BRACHIARIA DECUMBENS pasture, sabiá (MIMOSA CAESALPINIIFOLIA plantations and Atlantic Forest areas of the Dry Forest Zone of Pernambuco. A total of 1,575 isolates were obtained from 398 groups. A single random isolate of each group was authenticated, in randomized blocks with two replications. Each plant was inoculated with 1 mL of a bacterial broth, containing an estimated population of 10(8 rhizobial cells mL-1. Forty-five days after inoculation, the plants were harvested, separated into shoots, roots and nodules, oven-dried to constant mass, and weighed. Next, the symbiotic capability was tested with 1.5 kg of an autoclaved sand:vermiculite (1:1 mixture in polyethylene bags. The treatments consisted of 122 authenticated isolates, selected based on the shoot dry matter, five uninoculated controls (treated with 0, 50, 100, 150, or 200 kg ha-1 N and a control inoculated with SEMIA 6152 (=BR1602, a strain of BRADYRHIZOBIUM JAPONICUM The test was performed as described above. The shoot dry matter of the plants inoculated with the most effective isolates did not differ from that of plants treated with 150 kg ha-1 N. Shoot dry matter was positively correlated with all other variables. The proportion of effective isolates was highest among isolates from SABIÁ forests. There was great variation in nodule dry weight, as well as in N contents and total N.

  7. 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

  8. 根瘤菌培养基的优化和剂型的比较研究%MEDIUM OPTIMIZATION AND INOCULANT TYPE COMPARISON OF RHIZOBIUM

    Institute of Scientific and Technical Information of China (English)

    吴红慧; 周俊初

    2004-01-01

    以费氏中华根瘤菌(Sinorhizobium fredii)HN01和大豆慢生根瘤菌(Bradyrhizobium japonicum)USDAl10作为供试菌,进行YMA、TY、SM、PA和BSE等5种培养基的比较试验.结果表明,两种菌都在BSE中生长速度最快.对USDA110进行培养基的优化试验,筛选出一种能将生长速度提高2倍左右的优化培养基.制作了供试菌的固体、液体与冻干菌剂,结果表明在3种供试固体剂型载体中,草炭要优于蛭石,蛭石又优于珍珠岩.供试菌在两种液体剂型中的存活率和活菌数均较高,氮气或真空剂型对存活率的影响没有明显的差别.两种冻干菌剂的结果表明,供试菌在冻干过程中的死亡率较高,液氮冻干优于常规冻干.冻干菌剂在储存条件下的存活率为:-20℃>4℃>室温.

  9. Genomic analysis of cyclic-di-GMP-related genes in rhizobial type strains and functional analysis in Rhizobium etli.

    Science.gov (United States)

    Gao, Shanjun; Romdhane, Samir Ben; Beullens, Serge; Kaever, Volkhard; Lambrichts, Ivo; Fauvart, Maarten; Michiels, Jan

    2014-05-01

    Rhizobia are soil bacteria that can fix nitrogen in symbiosis with leguminous plants or exist free living in the rhizosphere. Crucial to their complex lifestyle is the ability to sense and respond to diverse environmental stimuli, requiring elaborate signaling pathways. In the majority of bacteria, the nucleotide-based second messenger cyclic diguanosine monophosphate (c-di-GMP) is involved in signal transduction. Surprisingly, little is known about the importance of c-di-GMP signaling in rhizobia. We have analyzed the genome sequences of six well-studied type species (Bradyrhizobium japonicum, Mesorhizobium loti, Rhizobium etli, Rhizobium leguminosarum, Sinorhizobium fredii, and Sinorhizobium meliloti) for proteins possibly involved in c-di-GMP signaling based on the presence of four domains: GGDEF (diguanylate cyclase), EAL and HD-GYP (phosphodiesterase), and PilZ (c-di-GMP sensor). We find that rhizobia possess a high number of these proteins. Conservation analysis suggests that c-di-GMP signaling proteins modulate species-specific pathways rather than ancient rhizobia-specific processes. Two hybrid GGDEF-EAL proteins were selected for functional analysis, R. etli RHE_PD00105 (CdgA) and RHE_PD00137 (CdgB). Expression of cdgA and cdgB is repressed by the alarmone (p)ppGpp. cdgB is significantly expressed on plant roots and free living. Mutation of cdgA, cdgB, or both does not affect plant root colonization, nitrogen fixation capacity, biofilm formation, motility, and exopolysaccharide production. However, heterologous expression of the individual GGDEF and EAL domains of each protein in Escherichia coli strongly suggests that CdgA and CdgB are bifunctional proteins, possessing both diguanylate cyclase and phosphodiesterase activities. Taken together, our results provide a platform for future studies of c-di-GMP signaling in rhizobia.

  10. Molecular and microscopic analysis of the gut contents of abundant rove beetle species (Coleoptera, Staphylinidae in the boreal balsam fir forest of Quebec, Canada

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    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

  11. 海金沙草化学成分的研究%Studies on the chemical constituents in herb of Lygodium japonicum

    Institute of Scientific and Technical Information of China (English)

    张雷红; 殷志琦; 叶文才; 赵守训; 王磊; 胡芳

    2005-01-01

    目的:对海金沙草的化学成分进行研究.方法:采用色谱技术进行分离,通过NMR等波谱方法确定化合物的结构.结果:分离并鉴定了8个化合物,分别为:田蓟苷(tilianin)(Ⅰ),山柰酚-7-O-α-L-吡喃鼠李糖苷(kaempferol-7-O-α-L-rhamnopyranoside)(Ⅱ),山柰酚(kaempferol)(Ⅲ),对香豆酸(p-coumaric acid)(Ⅳ),1-正十六烷酸甘油酯(hexadecanoic acid 2,3-出hydroxy-propyl ester)(Ⅴ),胡萝卜苷(daucosterol)(Ⅵ),β-谷甾醇(β-sitosterol)(Ⅶ),正三十一烷醇(1-hentriacontanol)(Ⅷ).结论:除化合物Ⅲ,Ⅳ外,其余均为首次从海金沙中分离得到.化合物Ⅰ,Ⅱ,Ⅴ为首次从海金沙属植物中分离得到.

  12. Genome Sequence of Pectobacterium carotovorum Phage PPWS1, Isolated from Japanese Horseradish [Eutrema japonicum (Miq.) Koidz] Showing Soft-Rot Symptoms.

    Science.gov (United States)

    Hirata, Hisae; Kashihara, Misako; Horiike, Tokumasa; Suzuki, Tomohiro; Dohra, Hideo; Netsu, Osamu; Tsuyumu, Shinji

    2016-04-21

    ITALIC! Pectobacterium carotovorumsubsp. ITALIC! carotovorumand its lytic bacteriophage PPWS1 were isolated from a Japanese horseradish rhizome with soft rot. Sequencing of the phage genomic DNA suggested that PPWS1 is a new species of the family ITALIC! Podoviridaeand has high similarity to the bacteriophage Peat1 infectious to ITALIC! P. atrosepticum.

  13. Dynamic observation of splenocyte apoptosis in mice immunized with recombinant vaccine Bifidobacterium bifidum pGEX-Sj14-3-3 of Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    张宁

    2013-01-01

    Objective To investigate the effects of recombinant vaccine Bifidobacterium bifidum(Bb) pGEX-Sj14-3-3 on splenocyte apoptosis in BALB/c mice. Methods Ninety-six BALB/c mice were randomly divided into two groups according to their body mass: per os group(PO) and

  14. Environ: E00359 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available E00359 Aconitum japonicum tuberous root Crude drug Aconitine [CPD:C06091], Mesaconi...amine [CPD:C06346], Coryneine chloride Aconitum japonicum, Aconitum [TAX:49188] Ranunculaceae (buttercup family) Aconitum... japonicum tuberous root Crude drugs [BR:br08305] Dicot plants: others Ranunculaceae (buttercup family) E00359 Aconitum tuberous root ...

  15. NCBI nr-aa BLAST: CBRC-MMUR-01-0199 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-0199 emb|CAX75788.1| Tubulin beta-2C chain [Schistosoma japonicum] emb...|CAX75790.1| Tubulin beta-2C chain [Schistosoma japonicum] emb|CAX75791.1| Tubulin beta-2C chain [Schistosoma japonicum] CAX75788.1 2e-31 82% ...

  16. Advances in Schistosoma japonicum recombinant protein glutathione-S-transferase(SjGST)and Schistosoma japonicum signaling protein 14-3-3(Sj14-3-3)%日本血吸虫GST蛋白和14-3-3蛋白的研究进展

    Institute of Scientific and Technical Information of China (English)

    刘文; 徐振山; 宋礼华

    2011-01-01

    Schistosomiasis is one of the serious health-threatening parasitic zoonoses to human beings. It has been presented in 76 countries and regions. More than 200 million people have been suffered from the infectious diseases all over the world. At present,some comprehensive precautionary and therapeutic measures were introduced to prevent schistosomiasis of human beings and animals simultaneously. The measures have brought some obvious positive effects. but the high cost combined with the high re-infection rate and such problems are still barriers to the application of the measures. Thus, to seek novel therapeutic medicines, candidates for the vaccines and also manage to develop standard immunologic diagnosis reagents with high specificity and sensitivity is the essential topic of the basic schistosomiasis research currently. The recent study of progress of the SjGST protein was reviewed, most potential candidates for the vaccines presented by WHO were summarized. Besides. this paper briefly reviewed the Sj14-3-3 protein which was involved in many functions of Schistosoma.%血吸虫病是严重危害人类健康的人兽共患病之一,在全球范围内,血吸虫病曾在76个国家和地区流行,有超过2亿人感染了血吸虫病.目前,世界上采取了一些人畜同步治疗等综合防治措施,虽取得了一定的成效,但仍面临着成本高、再感染率高等一系列问题.因此,寻找新的治疗药物、疫苗候选分子以及开发高度特异且敏感的标准化免疫诊断试剂是当前日本血吸虫病基础研究的重要内容.主要对WHO提出的最具潜力的疫苗候选分子血吸虫GST蛋白以及参与血吸虫许多生物学功能的14-3-3蛋白的最新研究进展进行一些阐述.

  17. 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

  18. 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

  19. 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

  20. Complete Genomic Structure of the Cultivated Rice Endophyte Azospirillum sp. B510

    Science.gov (United States)

    Kaneko, Takakazu; Minamisawa, Kiwamu; Isawa, Tsuyoshi; Nakatsukasa, Hiroki; Mitsui, Hisayuki; Kawaharada, Yasuyuki; Nakamura, Yasukazu; Watanabe, Akiko; Kawashima, Kumiko; Ono, Akiko; Shimizu, Yoshimi; Takahashi, Chika; Minami, Chiharu; Fujishiro, Tsunakazu; Kohara, Mitsuyo; Katoh, Midori; Nakazaki, Naomi; Nakayama, Shinobu; Yamada, Manabu; Tabata, Satoshi; Sato, Shusei

    2010-01-01

    We determined the nucleotide sequence of the entire genome of a diazotrophic endophyte, Azospirillum sp. B510. Strain B510 is an endophytic bacterium isolated from stems of rice plants (Oryza sativa cv. Nipponbare). The genome of B510 consisted of a single chromosome (3 311 395 bp) and six plasmids, designated as pAB510a (1 455 109 bp), pAB510b (723 779 bp), pAB510c (681 723 bp), pAB510d (628 837 bp), pAB510e (537 299 bp), and pAB510f (261 596 bp). The chromosome bears 2893 potential protein-encoding genes, two sets of rRNA gene clusters (rrns), and 45 tRNA genes representing 37 tRNA species. The genomes of the six plasmids contained a total of 3416 protein-encoding genes, seven sets of rrns, and 34 tRNAs representing 19 tRNA species. Eight genes for plasmid-specific tRNA species are located on either pAB510a or pAB510d. Two out of eight genomic islands are inserted in the plasmids, pAB510b and pAB510e, and one of the islands is inserted into trnfM-CAU in the rrn located on pAB510e. Genes other than the nif gene cluster that are involved in N2 fixation and are homologues of Bradyrhizobium japonicum USDA110 include fixABCX, fixNOQP, fixHIS, fixG, and fixLJK. Three putative plant hormone-related genes encoding tryptophan 2-monooxytenase (iaaM) and indole-3-acetaldehyde hydrolase (iaaH), which are involved in IAA biosynthesis, and ACC deaminase (acdS), which reduces ethylene levels, were identified. Multiple gene-clusters for tripartite ATP-independent periplasmic-transport systems and a diverse set of malic enzymes were identified, suggesting that B510 utilizes C4-dicarboxylate during its symbiotic relationship with the host plant. PMID:20047946

  1. 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

  2. Disponibilidade de potássio e suas relações com cálcio e magnésio em soja cultivada em casa-de-vegetação

    Directory of Open Access Journals (Sweden)

    Oliveira Fábio Alvares de

    2001-01-01

    Full Text Available A falta de resposta da soja à aplicação de K em elevadas quantidades pode estar relacionada com suas interações com o Ca e o Mg do solo e, portanto, com a calagem. O objetivo deste trabalho foi avaliar a nutrição potássica da soja em relação ao cálcio e ao magnésio do solo, por um experimento em vasos protegidos, sob casa-de-vegetação, utilizando-se um Latossolo Vermelho Amarelo distrófico típico, textura média, disposto num esquema fatorial 5x7 em blocos casualizados, com 4 repetições. Aplicaram-se, 40 dias antes da semeadura, cinco níveis de calcário dolomítico calcinado (0, 500, 1000, 1500, 2000 mg dm-3, combinados com sete níveis de K (0, 15, 30, 45, 60, 75, 90 mg dm-3, aplicados na forma de KCl, previamente à semeadura. Em cada vaso, cultivaram-se 4 plantas de soja inoculadas com Bradyrhizobium japonicum, cultivar precoce IAC-17, com 112 dias de ciclo. Avaliaram-se duas plantas de cada vaso no estádio R2 (florescimento pleno e duas no estádio R8 (maturação plena. Quando a relação (Ca+Mg/K trocável no solo foi superior a 36 ou a relação dos teores foliares (Ca+Mg/K superior a 3,6, foram verificadas as menores produções de material vegetal em plantas com sintomas de deficiência e teores foliares reduzidos de K. Por outro lado, as maiores produções, aliadas ao maior equilíbrio dos teores foliares de K, Ca e Mg foram obtidas quando a relação (Ca+Mg/K trocável no solo apresentava-se entre 20 e 30. A relação (Ca+Mg/K trocável no solo mostrou-se um índice importante de avaliação da disponibilidade do K no solo para a cultura da soja. A recomendação de adubação potássica para a cultura da soja também deve considerar a quantidade de calcário aplicada.

  3. RNA-Seq analysis of differential gene expression responding to different rhizobium strains in soybean (Glycine max roots

    Directory of Open Access Journals (Sweden)

    Songli eYuan

    2016-05-01

    Full Text Available The root nodule symbiosis (RNS between legume plants and rhizobia is the most efficient and productive source of nitrogen fixation, and has critical importance in agriculture and mesology. Soybean (Glycine max, one of the most important legume crops in the world, establishes a nitrogen-fixing symbiosis with different types of rhizobia, and the efficiency of symbiotic nitrogen fixation in soybean greatly depends on the symbiotic host-specificity. Although it has been reported that rhizobia use surface polysaccharides, secretion proteins of the type-three secretion systems and nod factors to modulate host range, the host control of nodulation specificity remains poorly understood. In this report, the soybean roots of two symbiotic systems (Bradyrhizobium japonicum strain 113-2-soybean and Sinorhizobium fredii USDA205-soybeanwith notable different nodulation phenotypes and the control were studied at five different post-inoculation time points (0.5h, 7-24h, 5d, 16d and 21d by RNA-seq (Quantification. The results of qPCR analysis of 11 randomly-selected genes agreed with transcriptional profile data for 136 out of 165 (82.42% data points and quality assessment showed that the sequencing library is of quality and reliable. Three comparisons (control vs 113-2, control vs USDA205 and USDA205 vs 113-2 were made and the differentially expressed genes (DEGs between them were analyzed. The number of DEGs at 16 days post-inoculation (dpi was the highest in the three comparisons, and most of the DEGs in USDA205 vs 113-2 were found at 16 dpi and 21 dpi. 44 go function terms in USDA205 vs 113-2 were analyzed to evaluate the potential functions of the DEGs, and 10 important KEGG pathway enrichment terms were analyzed in the three comparisons. Some important genes induced in response to different strains (113-2 and USDA205 were identified and analyzed, and these genes primarily encoded soybean resistance proteins, NF-related proteins, nodulins and immunity

  4. 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

  5. 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.

  6. 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.

  7. 日本血吸虫Calpain序列分析及DNA疫苗体系的构建%Sequence analysis and DNA vaccine preparation of Calpain,Schistosoma japonicum vaccine candidate molecule

    Institute of Scientific and Technical Information of China (English)

    周爱琴; 张仁利; 石淑华; 龙彩虹; 高世同; 林敏; 吴少庭

    2003-01-01

    目的探讨日本血吸虫疫苗候选分子-钙离子激活的中性蛋白激酶(Calpain)在抗日本血吸虫感染的保护性作用及其保护性免疫机制.方法从日本血吸虫成虫中提取RNA,用RT-PCR扩增Calpain含多个B,T细胞表位的片段,引物中包含BamHI和EcoRI的酶切位点,PCR扩增产物经纯化后TA克隆,转化的阳性TA克隆经PCR筛选后,液体培养大肠杆菌并回收质粒DNA,质粒DNA经BamHⅠ和EcoRⅠ双酶切,目的基因亚克隆到真核表达质粒pVAC载体,构建pVAC-Calpain真核表达体系,转化的阳性亚克隆经PCR筛选,液体培养大肠杆菌并回收pVAC-Calpain质粒DNA,质粒DNA经BamHⅠ和EcoRⅠ双酶切和DNA序列分析鉴定被亚克隆的Calpain基因.结果 RT-PCR从日本血吸虫RNA中扩增了453bp的Calpain基因,经BamHⅠ和EcoRⅠ双酶切和DNA序列分析鉴定Calpain基因被克隆到真核表达质粒pVAC载体.结论日本血吸虫疫苗候选分子Calpain的DNA疫苗体系的建立将有助于解析这个疫苗候选分子抗日本血吸虫感染的保护性免疫作用及保护性免疫机制.

  8. Recombinant protein of Schistosoma japonicum calpain induced antibody production and detection of schistosoma infection%重组Calpain蛋白诱导抗体产生及检测日本血吸虫感染的研究

    Institute of Scientific and Technical Information of China (English)

    张仁利; 吴少庭; 高世同; 黄达娜; 袁仕善

    2003-01-01

    目的研究在大肠杆菌中表达的日本血吸虫Calpain蛋白的免疫原性及其在日本血吸虫病诊断上的应用. 方法用重组Calpain蛋白免疫BALB/c小鼠,ELISA法测定特异性IgG抗体的动态变化及其IgG亚型(IgG1,IgG2a,IgG2b,IgG3)产生特征;同时用重组Calpain蛋白作为诊断抗原,粪检血吸虫病阳性患者血清作为一抗,肝吸虫阳性患者血清作为考核交叉反应血清. 结果重组Calpain抗原免疫小鼠后,产生了一个极高的抗Calpain特异性抗体,免疫4周IgG类抗体达到高峰,与对照组鼠相比较免疫鼠血清中IgG1,IgG2a,IgG2b特异性抗体也显著上升;Calpain重组抗原血清检测日本血吸虫和粪检的符合率为100%,粪检阳性EPG低的个体抗Calpain抗体滴度高,EPG高的患者抗Calpain抗体滴度反而低,与肝吸虫的交叉反应率为37%. 结论日本血吸虫Calpain是一个具有免疫原性的蛋白,能够激发宿主产生高水平的免疫球蛋白,能够敏感地测定日本血吸虫的感染,提示Calpain可以发展成为日本血吸虫病的诊断抗原和日本血吸虫疫苗候选分子.

  9. 日本血吸虫Calpain疫苗候选分子抗感染的免疫机制%Immunoprotective Mechanism of Calpain as Vaccine Candidate against Schistosoma japonicum Infection

    Institute of Scientific and Technical Information of China (English)

    张仁利; 黄达娜; 高世同; 耿艺介; 吴少庭; 胡章立; 张顺祥;