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Sample records for juncea recombinant lines

  1. A duplicated coxI gene is associated with cytoplasmic male sterility in an alloplasmic Brassica juncea line derived from somatic hybridization with Diplotaxis catholica

    Indian Academy of Sciences (India)

    Aruna Pathania; Rajesh Kumar; V. Dinesh Kumar; Ashutosh; K. K. Dwivedi; P. B. Kirti; P. Prakash; V. L. Chopra; S. R. Bhat

    2007-08-01

    A cytoplasmic male sterile (CMS) line of Brassica juncea was derived by repeated backcrossing of the somatic hybrid (Diplotaxis catholica + B. juncea) to B. juncea. The new CMS line is comparable to euplasmic lines for almost all characters, except for flowers which bear slender, needle-like anthers with aborted pollen. Detailed Southern analysis revealed two copies of coxI gene in the CMS line. One copy, coxI-1 is similar to the coxI gene of B. juncea, whereas the second copy, coxI-2 is present in a novel rearranged region. Northern analysis with eight mitochondrial gene probes showed altered transcript pattern only for the coxI gene. Two transcripts of 2.0 and 2.4 kb, respectively, were detected in the CMS line. The novel 2.4 kb transcript was present in floral bud tissue but absent in the leaf tissue. In plants where male sterility broke down under high temperature during the later part of the growing season, the 2.4 kb coxI transcript was absent, which suggested its association with the CMS. The two coxI genes from the CMS line showed two amino acid changes in the coding region. The novel coxI gene showed unique repeats in the 5′ region suggesting recombination of mitochondrial genomes of the two species. The possible role of the duplicated coxI gene in causing male sterility is discussed.

  2. Brassica juncea Lines with Substituted Chimeric GFP-CENH3 Give Haploid and Aneuploid Progenies on Crossing with Other Lines

    OpenAIRE

    Watts, Anshul; Singh, Sunil K.; Bhadouria, Jyoti; Naresh, Vasupalli; Bishoyi, Ashok K.; K.A. Geetha; Chamola, Rohit; Pattanayak, Debasis; Bhat, Shripad R.

    2017-01-01

    Haploids and doubled haploids are invaluable for basic genetic studies and in crop improvement. A novel method of haploid induction through genetic engineering of the Centromere Histone Protein gene, CENH3, has been demonstrated in Arabidopsis. The present study was undertaken to develop haploid inducer (HI) lines of Brassica juncea based on the principles elaborated in Arabidopsis. B. juncea was found to carry three copies of CENH3 which generated five different transcripts, of which three t...

  3. Development and characterization of Brassica juncea – fruticulosa introgression lines exhibiting resistance to mustard aphid (Lipaphis erysimi Kalt

    Directory of Open Access Journals (Sweden)

    Atri Chhaya

    2012-11-01

    Full Text Available Abstract Background Mustard aphid is a major pest of Brassica oilseeds. No source for aphid resistance is presently available in Brassica juncea . A wild crucifer, Brassica fruticulosa is known to be resistant to mustard aphid. An artificially synthesized amphiploid, AD-4 (B. fruticulosa × B. rapa var. brown sarson was developed for use as a bridge species to transfer fruticulosa resistance to B. juncea. Using the selfed backcross we could select a large number of lines with resistance to mustard aphid. This paper reports cytogenetic stability of introgression lines, molecular evidence for alien introgression and their reaction to mustard aphid infestation. Results Majority of introgression lines had expected euploid chromosome number(2n= 36, showed normal meiosis and high pollen grain fertility. Well-distributed and transferable simple-sequence repeats (SSR markers for all the 18 B. juncea chromosomes helped to characterize introgression events. Average proportions of recipient and donor genome in the substitution lines were 49.72 and 35.06%, respectively. Minimum alien parent genome presence (27.29% was observed in the introgression line, Ad3K-280 . Introgressed genotypes also varied for their resistance responses to mustard aphid infestations under artificial release conditions for two continuous seasons. Some of the test genotypes showed consistent resistant reaction. Conclusions B.juncea-fruticulosa introgression set may prove to be a very powerful breeding tool for aphid resistance related QTL/gene discovery and fine mapping of the desired genes/QTLs to facilitate marker assisted transfer of identified gene(s for mustard aphid resistance in the background of commercial mustard genotypes.

  4. Radio Recombination Lines of Hydrogen

    Indian Academy of Sciences (India)

    G. Peach

    2015-12-01

    The impact theory of spectral line broadening is used to obtain complete profiles for radio recombination lines perturbed by electron and proton impact. The collisions can be divided into two types: inelastic, where transitions take place between hydrogen levels with different principal quantum number and elastic, where the transitions are only between degenerate levels for a particular value of . The widths of the radio lines are essentially determined by inelastic electron collisions and elastic proton collisions with the emitting hydrogen atom occupying either the upper or lower levels of the line. Here, earlier work is extended to examine the contribution from proton collisions to the line width in more detail, and it is shown that the trends in the behaviour of the widths again confirm previous results.

  5. Workshop on Radio Recombination Lines

    CERN Document Server

    1980-01-01

    Since their first detection 15 years ago, radio recombination lines from several elements have been observed in a wide variety of objects including HII regions, planetary nebulae, molecular clouds, the diffuse interstellar medium, and recently, other galaxies. The observations span almost the entire range from 0.1 to 100 GHz, and employ both single­ djsh and aperture synthesis techniques. The theory of radio recombination lines has also advanced strongly, to the point where it is perhaps one of the best-understood in astro­ physics. In a parallel development, it has become possible over the last decade to study these same highly-excited atoms in the laboratory; this work provides further confirmation of the theoretical framework. However there has been continuing controversy over the astrophysical interpre­ tation of radio recombination line observations, especially regarding the role of stimulated emission. A workshop was held in Ottawa on 24-25 August, 1979, bringing together many of the active scientist...

  6. Theoretic Study of CⅡ Recombination Line

    Institute of Scientific and Technical Information of China (English)

    彭永伦; 王民盛; 韩小英; 李家明

    2004-01-01

    Using the R-matrix method, we carry out theoretical calculations for recombination line λ 8794 A(3d'-3p') of CⅡ, which is important to estimate the abundances of carbon in planetary nebulae. Our calculations are based on three sets of target orbital basis, through which we elucidate the electron correlation and static polarization effects in the dielectronic recombination processes.

  7. Radio Recombination Lines in Galactic HII Regions

    CERN Document Server

    Quireza, C; Bania, T M; Rood, R T; Balser, Dana S.; Quireza, Cintia; Rood, Robert T.

    2006-01-01

    We report radio recombination line (RRL) and continuum observations of a sample of 106 Galactic HII regions made with the NRAO 140 Foot radio telescope in Green Bank, WV. We believe this to be the most sensitive RRL survey ever made for a sample this large. Most of our source integration times range between 6 and 90 hours which yield typical r.m.s. noise levels of 1.0--3.5 milliKelvins. Our data result from two different experiments performed, calibrated, and analyzed in similar ways. A CII survey was made at 3.5 cm wavelength to obtain accurate measurements of carbon radio recombination lines. When combined with atomic (CI) and molecular (CO) data, these measurements will constrain the composition, structure, kinematics, and physical properties of the photodissociation regions that lie on the edges of HII regions. A second survey was made at 3.5 cm wavelength to determine the abundance of 3He in the interstellar medium of the Milky Way. Together with measurements of the 3He+ hyperfine line we get high precis...

  8. An evolutionarily conserved mitochondrial orf108 is associated with cytoplasmic male sterility in different alloplasmic lines of Brassica juncea and induces male sterility in transgenic Arabidopsis thaliana.

    Science.gov (United States)

    Kumar, Pankaj; Vasupalli, Naresh; Srinivasan, R; Bhat, Shripad R

    2012-05-01

    Nuclear-mitochondrial gene interactions governing cytoplasmic male sterility (CMS) in angiosperms have been found to be unique to each system. Fertility restoration of three diverse alloplasmic CMS lines of Brassica juncea by a line carrying the fertility-restorer gene introgressed from Moricandia arvensis prompted this investigation to examine the molecular basis of CMS in these lines. Since previous studies had found altered atpA transcription associated with CMS in these lines, the atpA genes and transcripts of CMS, fertility-restored, and euplasmic lines were cloned and compared. atpA coding and downstream sequences were conserved among CMS and euplasmic lines but major differences were found in the 5' flanking sequences of atpA. A unique open reading frame (ORF), orf108, co-transcribed with atpA, was found in male sterile flowers of CMS lines carrying mitochondrial genomes of Diplotaxis berthautii, D. catholica, or D. erucoides. In presence of the restorer gene, the bicistronic orf108-atpA transcript was cleaved within orf108 to yield a monocistronic atpA transcript. Transgenic expression of orf108 with anther-specific Atprx18 promoter in Arabidopsis thaliana gave 50% pollen sterility, indicating that Orf108 is lethal at the gametophytic stage. Further, lack of transmission of orf108 to the progeny showed for the first time that mitochondrial ORFs could also cause female sterility. orf108 was found to be widely distributed among wild relatives of Brassica, indicating its ancient origin. This is the first report that shows that CMS lines of different origin and morphology could share common molecular basis. The gametic lethality of Orf108 offers a novel opportunity for transgene containment.

  9. Homeotic-like modification of stamens to petals is associated with aberrant mitochondrial gene expression in cytoplasmic male sterile Ogura Brassica juncea

    Indian Academy of Sciences (India)

    Gargi Meur; K. Gaikwad; S. R. Bhat; S. Prakash; P. B. Kirti

    2006-08-01

    We have previously reported correction of severe leaf chlorosis in the cytoplasmic male sterile Ogura (also called Ogu) Brassica juncea line carrying Ogura cytoplasm by plastid substitution via protoplast fusion. Two cybrids obtained from the fusion experiment, Og1 and Og2, were green and carried the plastid genome of B. juncea cv. RLM198. While Og1 displayed normal flower morphology comparable to that of its euplasmic B. juncea counterpart except for sterile anthers, Og2 retained homeotic-like floral modification of stamens to petal-like structures and several other floral deformities observed in the chlorotic (Ogu) B. juncea cv. RLM198 (or OgRLM). With respect to the mitochondrial genome, Og1 showed 81% genetic similarity to the fertile cultivar RLM while Og2 showed 93% similarity to OgRLM. In spite of recombination and rearrangements in the mitochondrial genomes in the cybrids, expression patterns of 10 out of 11 mitochondrial genes were similar in all the three CMS lines; the only exception was atp6, whose expression was altered. While Og1 showed normal atp6 transcript similar to that in RLM, in Og2 and OgRLM weak expression of a longer transcript was detected. These results suggest that the homeotic-like changes in floral patterning leading to petaloid stamens in Og2 and OgRLM may be associated with aberrant mitochondrial gene expression.

  10. Caenorhabditis briggsae recombinant inbred line genotypes reveal inter-strain incompatibility and the evolution of recombination.

    Directory of Open Access Journals (Sweden)

    Joseph A Ross

    2011-07-01

    Full Text Available The nematode Caenorhabditis briggsae is an emerging model organism that allows evolutionary comparisons with C. elegans and exploration of its own unique biological attributes. To produce a high-resolution C. briggsae recombination map, recombinant inbred lines were generated from reciprocal crosses between two strains and genotyped at over 1,000 loci. A second set of recombinant inbred lines involving a third strain was also genotyped at lower resolution. The resulting recombination maps exhibit discrete domains of high and low recombination, as in C. elegans, indicating these are a general feature of Caenorhabditis species. The proportion of a chromosome's physical size occupied by the central, low-recombination domain is highly correlated between species. However, the C. briggsae intra-species comparison reveals striking variation in the distribution of recombination between domains. Hybrid lines made with the more divergent pair of strains also exhibit pervasive marker transmission ratio distortion, evidence of selection acting on hybrid genotypes. The strongest effect, on chromosome III, is explained by a developmental delay phenotype exhibited by some hybrid F2 animals. In addition, on chromosomes IV and V, cross direction-specific biases towards one parental genotype suggest the existence of cytonuclear epistatic interactions. These interactions are discussed in relation to surprising mitochondrial genome polymorphism in C. briggsae, evidence that the two strains diverged in allopatry, the potential for local adaptation, and the evolution of Dobzhansky-Muller incompatibilities. The genetic and genomic resources resulting from this work will support future efforts to understand inter-strain divergence as well as facilitate studies of gene function, natural variation, and the evolution of recombination in Caenorhabditis nematodes.

  11. Recombinant protein production from stable mammalian cell lines and pools.

    Science.gov (United States)

    Hacker, David L; Balasubramanian, Sowmya

    2016-06-01

    We highlight recent developments for the production of recombinant proteins from suspension-adapted mammalian cell lines. We discuss the generation of stable cell lines using transposons and lentivirus vectors (non-targeted transgene integration) and site-specific recombinases (targeted transgene integration). Each of these methods results in the generation of cell lines with protein yields that are generally superior to those achievable through classical plasmid transfection that depends on the integration of the transfected DNA by non-homologous DNA end-joining. This is the main reason why these techniques can also be used for the generation of stable cell pools, heterogenous populations of recombinant cells generated by gene delivery and genetic selection without resorting to single cell cloning. This allows the time line from gene transfer to protein production to be reduced.

  12. Registration of 'RU9101001'/'Katy' recombinant inbred lines of rice

    Science.gov (United States)

    The cross of RU9101001/'Katy' rice (Oryza sativa L.) was used to develop a mapping population consisting of 238 F9 generation recombinant inbred lines of rice (Oryza sativa L.) (GSOR100361 to GSOR100600). This population has been used to map major genes that provide resistance to the rice blast pat...

  13. Radio recombination lines from obscured quasars with the SKA

    CERN Document Server

    Manti, Serena; Ferrara, Andrea; Feruglio, Chiara; Graziani, Luca; Bernardi, Gianni

    2015-01-01

    We explore the possibility of detecting hydrogen radio recombination lines from 0 < z < 10 quasars. We compute the expected Hnalpha flux densities as a function of absolute magnitude and redshift by considering (i) the range of observed AGN spectral indices from UV to X-ray bands, (ii) secondary ionizations from X-ray photons, and (iii) stimulated emission due to nonthermal radiation. All these effects are important to determine the line fluxes. We find that the combination of slopes: alpha_X,hard = -1.11, alpha_X,soft = -0.7, alpha_EUV = -1.3, alpha_UV = -1.7, maximizes the expected flux, f_Hnalpha = 10 microJy for z = 7 quasars with M_AB = -27 in the n = 50 lines; allowed SED variations produce variations by a factor of 3 around this value. Secondaries boost the line intensity by a factor of 2 to 4, while stimulated emission in high-z quasars with M_AB = -26 provides an extra boost to RRL flux observed at nu = 1 GHz if recombinations arise in HII regions with T_e = 10^3-5 K, n_e = 10^3-5 cm^-3. We com...

  14. Transfer line from the PSB to the PS (recombination)

    CERN Multimedia

    1976-01-01

    After sequential ejection of 5 bunches from each of the 4 rings of the Booster (originally 800 MeV, now 1.4 GeV), the 4 batches are brought to the same vertical level, so as to form a string of 20 bunches, filling the circumference of the PS. This vertical "recombination" is performed in the transfer line, using vertical bending magnets, septa and kickers. Here we see the section where the beam from ring 4 (the top one) is brought down to the level of ring 3, and the beam from ring 1 up to the level of ring 2. Further downstream (to the right, outside this picture), level 2 is brought up to level 3, identical to that of the PS. After this original recombination scheme, other ways of combining the 4 beams, vertically and/or longitudinally, were developed and used in operation.

  15. Radio recombination lines from the largest bound atoms in space

    CERN Document Server

    Stepkin, S V; Kantharia, N G; Shankar, N U

    2006-01-01

    In this paper, we report the detection of a series of radio recombination lines (RRLs) in absorption near 26 MHz arising from the largest bound carbon atoms detected in space. These atoms, which are more than a million times larger than the ground state atoms are undergoing delta transitions (n~1009, Delta n=4) in the cool tenuous medium located in the Perseus arm in front of the supernova remnant, Cassiopeia A. Theoretical estimates had shown that atoms which recombined in tenuous media are stable up to quantum levels n~1500. Our data indicates that we have detected radiation from atoms in states very close to this theoretical limit. We also report high signal-to-noise detections of alpha, beta and gamma transitions in carbon atoms arising in the same clouds. In these data, we find that the increase in line widths with quantum number (proportional to n^5) due to pressure and radiation broadening of lines is much gentler than expected from existing models which assume a power law background radiation field. T...

  16. Human cell lines: A promising alternative for recombinant FIX production.

    Science.gov (United States)

    de Sousa Bomfim, Aline; Cristina Corrêa de Freitas, Marcela; Picanço-Castro, Virgínia; de Abreu Soares Neto, Mário; Swiech, Kamilla; Tadeu Covas, Dimas; Maria de Sousa Russo, Elisa

    2016-05-01

    Factor IX (FIX) is a vitamin K-dependent protein, and it has become a valuable pharmaceutical in the Hemophilia B treatment. We evaluated the potential of recombinant human FIX (rhFIX) expression in 293T and SK-Hep-1 human cell lines. SK-Hep-1-FIX cells produced higher levels of biologically active protein. The growth profile of 293T-FIX cells was not influenced by lentiviral integration number into the cellular genome. SK-Hep-1-FIX cells showed a significantly lower growth rate than SK-Hep-1 cells. γ-carboxylation process is significant to FIX biological activity, thus we performed a expression analysis of genes involved in this process. The 293T gene expression suggests that this cell line could efficiently carboxylate FIX, however only 28% of the total secreted protein is active. SK-Hep-1 cells did not express high amounts of VKORC1 and carboxylase, but this cell line secreted large amounts of active protein. Enrichment of culture medium with Ca(+2) and Mg(+2) ions did not affect positively rhFIX expression in SK-Hep-1 cells. In 293T cells, the addition of 0.5 mM Ca(+2) and 1 mM Mg(+2) resulted in higher rhFIX concentration. SK-Hep-1 cell line proved to be very effective in rhFIX production, and it can be used as a novel biotechnological platform for the production of recombinant proteins.

  17. DISCOVERY OF CARBON RADIO RECOMBINATION LINES IN M82

    Energy Technology Data Exchange (ETDEWEB)

    Morabito, Leah K.; Oonk, J. B. R.; Salgado, Francisco; Röttgering, H. J. A.; Tielens, A. G. G. M.; Haverkorn, Marijke [Leiden Observatory, Leiden University, P.O. Box 9513, 2300 RA Leiden (Netherlands); Toribio, M. Carmen; Heald, George [Netherlands Institute for Radio Astronomy (ASTRON), Postbus 2, 7990 AA Dwingeloo (Netherlands); Beck, Rainer; Adebahr, Björn [MPI für Radioastronomie, Auf dem Hügel 69, D-53121 Bonn (Germany); Best, Philip [SUPA, Institute for Astronomy, Royal Observatory Edinburgh, Blackford Hill, Edinburgh EH9 3HJ (United Kingdom); Beswick, Robert [Jodrell Bank Centre for Astrophysics, University of Manchester, Oxford Road, Manchester M13 9PL (United Kingdom); Bonafede, Annalisa; Brüggen, Marcus [Hamburg Observatory, University of Hamburg, Gojenbergsweg 112, D-21029 Hamburg (Germany); Brunetti, Gianfranco [INAF-Istituto di Radioastronomia, Via P. Gobetti 101, I-40129 Bologna (Italy); Chyży, Krzysztof T. [Obserwatorium Astronomiczne Uniwersytetu Jagiellońskiego, ul. Orla 171, 30-244 Kraków (Poland); Conway, J. E.; Horellou, Cathy [Department of Earth and Space Sciences, Chalmers University of Technology, Onsala Space Observatory, SE-43992 Onsala (Sweden); Van Driel, Wim [GEPI, Observatoire de Paris, UMR 8111, CNRS, Université Paris Diderot, 5 Place Jules Janssen, F-92190 Meudon (France); Gregson, Jonathan, E-mail: morabito@strw.leidenuniv.nl [Department of Physical Sciences, The Open University, Milton Keynes MK7 6AA (United Kingdom); and others

    2014-11-10

    Carbon radio recombination lines (RRLs) at low frequencies (≲ 500 MHz) trace the cold, diffuse phase of the interstellar medium, which is otherwise difficult to observe. We present the detection of carbon RRLs in absorption in M82 with the Low Frequency Array in the frequency range of 48-64 MHz. This is the first extragalactic detection of RRLs from a species other than hydrogen, and below 1 GHz. Since the carbon RRLs are not detected individually, we cross-correlated the observed spectrum with a template spectrum of carbon RRLs to determine a radial velocity of 219 km s{sup –1}. Using this radial velocity, we stack 22 carbon-α transitions from quantum levels n = 468-508 to achieve an 8.5σ detection. The absorption line profile exhibits a narrow feature with peak optical depth of 3 × 10{sup –3} and FWHM of 31 km s{sup –1}. Closer inspection suggests that the narrow feature is superimposed on a broad, shallow component. The total line profile appears to be correlated with the 21 cm H I line profile reconstructed from H I absorption in the direction of supernova remnants in the nucleus. The narrow width and centroid velocity of the feature suggests that it is associated with the nuclear starburst region. It is therefore likely that the carbon RRLs are associated with cold atomic gas in the direction of the nucleus of M82.

  18. Using crossover breakpoints in recombinant inbred lines to identify quantitative trait loci controlling the global recombination frequency.

    Science.gov (United States)

    Esch, Elisabeth; Szymaniak, Jessica M; Yates, Heather; Pawlowski, Wojciech P; Buckler, Edward S

    2007-11-01

    Recombination is a crucial component of evolution and breeding, producing new genetic combinations on which selection can act. Rates of recombination vary tremendously, not only between species but also within species and for specific chromosomal segments. In this study, by examining recombination events captured in recombinant inbred mapping populations previously created for maize, wheat, Arabidopsis, and mouse, we demonstrate that substantial variation exists for genomewide crossover rates in both outcrossed and inbred plant and animal species. We also identify quantitative trait loci (QTL) that control this variation. The method that we developed and employed here holds promise for elucidating factors that regulate meiotic recombination and for creation of hyperrecombinogenic lines, which can help overcome limited recombination that hampers breeding progress.

  19. Detection of Voigt Spectral Line Profiles of Hydrogen Radio Recombination Lines toward Sagittarius B2(N)

    CERN Document Server

    von Prochazka, Azrael A; Balser, Dana S; Ryans, Robert S I; Marshall, Adele H; Schwab, Fredric R; Hollis, Jan M; Jewell, Philip R; Lovas, Frank J

    2010-01-01

    We report the detection of Voigt spectral line profiles of radio recombination lines (RRLs) toward Sagittarius B2(N) with the 100-m Green Bank Telescope (GBT). At radio wavelengths, astronomical spectra are highly populated with RRLs, which serve as ideal probes of the physical conditions in molecular cloud complexes. An analysis of the Hn(alpha) lines presented herein shows that RRLs of higher principal quantum number (n>90) are generally divergent from their expected Gaussian profiles and, moreover, are well described by their respective Voigt profiles. This is in agreement with the theory that spectral lines experience pressure broadening as a result of electron collisions at lower radio frequencies. Given the inherent technical difficulties regarding the detection and profiling of true RRL wing spans and shapes, it is crucial that the observing instrumentation produce flat baselines as well as high sensitivity, high resolution data. The GBT has demonstrated its capabilities regarding all of these aspects,...

  20. Radio Recombination Lines at Decametre Wavelengths: Prospects for the Future

    CERN Document Server

    Peters, W M; Clarke, T E; Erickson, W C; Kassim, N E

    2010-01-01

    This paper considers the suitability of a number of emerging and future instruments for the study of radio recombination lines (RRLs) at frequencies below 200 MHz. These lines arise only in low-density regions of the ionized interstellar medium, and they may represent a frequency-dependent foreground for next-generation experiments trying to detect H I signals from the Epoch of Reionization and Dark Ages ("21-cm cosmology"). We summarize existing decametre-wavelength observations of RRLs, which have detected only carbon RRLs. We then show that, for an interferometric array, the primary instrumental factor limiting detection and study of the RRLs is the areal filling factor of the array. We consider the Long Wavelength Array (LWA-1), the LOw Frequency ARray (LOFAR), the low-frequency component of the Square Kilometre Array (SKA-lo), and a future Lunar Radio Array (LRA), all of which will operate at decametre wavelengths. These arrays offer digital signal processing, which should produce more stable and better ...

  1. Mitochondrial atpA gene is altered in a new orf220-type cytoplasmic male-sterile line of stem mustard (Brassica juncea).

    Science.gov (United States)

    Yang, Jing-Hua; Huai, Yan; Zhang, Ming-Fang

    2009-02-01

    The purpose of this research is to identify the probable mitochondrial factor associated with cytoplasmic male sterility (cms) by comparative analysis of cms and its isogenic maintainer lines in stem mustards. Dramatic variations in the morphology of floral organs were observed in cms stem mustard. Mitochondrial atpA gene was shown to be altered in cms compared with that in its maintainer line, of which mitochondrial atpA gene from its maintainer line was sequenced to encode 507 amino acids. It was indicative of high homology with mitochondrial atpA genes from other species, even as high as 94% in similarity with Oryza sativa in terms of amino acid constituents. However, only 429 amino acids were deduced in cms showing 83% similarity with atpA gene from its maintainer line. Two copies were observed in its maintainer line, but only one was found in cms. Such numerous differences of mitochondrial atpA gene between cms and its maintainer lines may not be the results of evolutionary divergence but the rearrangements of mitochondria. Expression of mitochondrial atpA gene was shown to be down-regulated in cms by using Northern blot. Consequently, mitochondrial ATP synthesis was severely decreased more than one fold in cms stem mustard indicating deficiency in mitochondrial ATP synthesis in this type of cms. Therefore, we deduced that mitochondrial atpA gene altered in cms could be associated with male-sterility in this type of cms.

  2. Alterations of RNA Editing for the Mitochondrial ATP9 Gene in a New orf220-type Cytoplasmic Male-sterile Line of Stem Mustard (Brassica juncea var. tumida)

    Institute of Scientific and Technical Information of China (English)

    Jing-Hua Yang; Ming-Fang Zhang; Jing-Quan Yu

    2007-01-01

    RNA editing for the mitochondrial ATP9 gene of encoding regions has been observed in both cytoplasmic malesterile and maintainer lines of stem mustard, where its editing capacity varied spatially and temporally in the cytoplasmic male sterility (CMS) line. There were four RNA editing sites for the mitochondrial ATP9 gene according to its normal editing sites in mustard, of which three sites occurred as C-to-U changes and one as a U-to-C change.As a result, the hydrophobicity of deduced ATP9 protein was reduced due to the conversions at its 17th, 45th and 64th positions. Meanwhile, the conservation of deduced ATP9 protein was enhanced by changes at the 56th position.Loss of a specific editing site for ATP9 was observed in juvenile roots, senile roots, senile leaves and floret buds of the CMS line. Comparatively, complete RNA editing for ATP9 gene was retained in juvenile roots, juvenile leaves and floret buds of its maintainer line; however, the loss of a specific editing site for ATP9 gene occurred at senile roots and senile leaves in its maintainer line. These observations allow us to produce a hypothesis that the dysfunction of a specific mitochondrisl gene arising from RNA editing could probably be a factor triggering CMS and organ senescence through unknown cross-talk pathways during development.

  3. Comparative pharmacology of a new recombinant FSH expressed by a human cell Line

    DEFF Research Database (Denmark)

    Koechling, Wolfgang; Plaksin, Daniel; Croston, Glenn

    2017-01-01

    Recombinant FSH proteins are important therapeutic agents for the treatment of infertility, including follitropin alfa expressed in Chinese Hamster Ovary (CHO) cells and, more recently, follitropin delta expressed in the human cell line PER.C6. These recombinant FSH proteins have distinct...

  4. Radio recombination lines from the supernova remnant candidate G3392-04

    CERN Document Server

    Shaver, P A; McGee, R X; Urdin, P G

    1980-01-01

    H109 alpha and H137 beta recombination lines have been detected from the direction of the supernova remnant candidate G339.2-0.4. The source appears to be a low-excitation H II region with an electron temperature of approximately 5000K and a considerable overabundance of heavy elements. The recombination lines which Manchester and Mebold (1977) detected in the direction of the nearby pulsar PSR 1641-45 may also arise in this H II region. (9 refs).

  5. Introgressing Subgenome Components from Brassica rapa and B. carinata to B. juncea for Broadening Its Genetic Base and Exploring Intersubgenomic Heterosis

    Science.gov (United States)

    Wei, Zili; Wang, Meng; Chang, Shihao; Wu, Chao; Liu, Peifa; Meng, Jinling; Zou, Jun

    2016-01-01

    Brassica juncea (AjAjBjBj), is an allotetraploid that arose from two diploid species, B. rapa (ArAr) and B. nigra (BnBn). It is an old oilseed crop with unique favorable traits, but the genetic improvement on this species is limited. We developed an approach to broaden its genetic base within several generations by intensive selection. The Ar subgenome from the Asian oil crop B. rapa (ArAr) and the Bc subgenome from the African oil crop B. carinata (BcBcCcCc) were combined in a synthesized allohexaploid (ArArBcBcCcCc), which was crossed with traditional B. juncea to generate pentaploid F1 hybrids (ArAjBcBjCc), with subsequent self-pollination to obtain newly synthesized B. juncea (Ar/jAr/jBc/jBc/j). After intensive cytological screening and phenotypic selection of fertility and agronomic traits, a population of new-type B. juncea was obtained and was found to be genetically stable at the F6 generation. The new-type B. juncea possesses good fertility and rich genetic diversity and is distinctly divergent but not isolated from traditional B. juncea, as revealed by population genetic analysis with molecular markers. More than half of its genome was modified, showing exotic introgression and novel variation. In addition to the improvement in some traits of the new-type B. juncea lines, a considerable potential for heterosis was observed in inter-subgenomic hybrids between new-type B. juncea lines and traditional B. juncea accessions. The new-type B. juncea exhibited a stable chromosome number and a novel genome composition through multiple generations, providing insight into how to significantly broaden the genetic base of crops with subgenome introgression from their related species and the potential of exploring inter-subgenomic heterosis for hybrid breeding. PMID:27909440

  6. Introgressing subgenome components from Brassica rapa and B. carinata to B. juncea for broadening its genetic base and exploring intersubgenomic heterosis

    Directory of Open Access Journals (Sweden)

    Zili Wei

    2016-11-01

    Full Text Available Brassica juncea (AjAjBjBj, is an allotetraploid that arose from two diploid species, B. rapa (ArAr and B. nigra (BnBn. It is an old oilseed crop with unique favorable traits, but the genetic improvement on this species is limited. We developed an approach to broaden its genetic base within several generations by intensive selection. The Ar subgenome from the Asian oil crop B. rapa (ArAr and the Bc subgenome from the African oil crop B. carinata (BcBcCcCc were combined in a synthesized allohexaploid (ArArBcBcCcCc, which was crossed with traditional B. juncea to generate pentaploid F1 hybrids (ArAjBcBjCc, with subsequent self-pollination to obtain newly synthesized B. juncea (Ar/jAr/jBc/jBc/j. After intensive cytological screening and phenotypic selection of fertility and agronomic traits, a population of new-type B. juncea was obtained and was found to be genetically stable at the F6 generation. The new-type B. juncea possesses good fertility and rich genetic diversity and is distinctly divergent but not isolated from traditional B. juncea, as revealed by population genetic analysis with molecular markers. More than half of its genome was modified, showing exotic introgression and novel variation. In addition to the improvement in some traits of the new-type B. juncea lines, a considerable potential for heterosis was observed in inter-subgenomic hybrids between new-type B. juncea lines and traditional B. juncea accessions. The new-type B. juncea exhibited a stable chromosome number and a novel genome composition through multiple generations, providing insight into how to significantly broaden the genetic base of crops with subgenome introgression from their related species and the potential of exploring inter-subgenomic heterosis for hybrid breeding.

  7. Expression of recombination-activating genes and T cell receptor gene recombination in the human T cell leukemia cell line.

    Science.gov (United States)

    Zou, Hong-yun; Ma, Li; Meng, Min-jie; Yao, Xin-sheng; Lin, Ying; Wu, Zhen-qiang; He, Xiao-wei; Wang, Ju-fang; Wang, Xiao-ning

    2007-03-05

    Recent studies have suggested that mature T cells can change their specificity through reexpression of recombination-activating genes (RAG) and RAG-mediated V(D)J recombination. This process is named receptor revision and has been observed in mature peripheral T cells from transgenic mice and human donors. However, whether thebreceptor revision in mature T cells is a random or orientated process remains poorly understood. Here we used the Jurkathuman T cell line, which represents a mature stage of T cell development, as a model to investigate the regulation of Tcell receptor (TCR) gene recombination. TCR Dbeta-Jbeta signal joint T cell receptor excision DNA circles (sjTRECs) were determined by nested and seminested PCR. Double-strand DNA breaks at recombination signal sequences (RSSs) in the TCRVbeta chain locus were detected by ligation-mediated-PCR. Further analysis of the complementarity-determining region 3 (CDR3) size of the TCRVbeta chain was examined by the TCR GeneScan technique. RAG1, RAG2, and three crucial components of the nonhomologous DNA end-joining (NHEJ) pathway were readily detected in Jurkat. Characteristics of junctional diversity of Dbeta2-Jbeta2 signal joints and ds RSS breaks associated with the Dbeta2 5' and Dbeta 2 3' sites were detected in DNA from Jurkat cells. CDR3 size and the gene sequences of the TCRVbeta chain did not change during cell proliferation. RAG1 and RAG2 and ongoing TCR gene recombination are coexpressed in Jurkat cells, but the ongoing recombination process may not play a role in modification of the TCR repertoire.However, the results suggest that Jurkat could be used as a model for studying the regulation of RAGs and V(D)J recombination and as a "special" model of the coexistence of TCR gene rearrangements and "negative" receptor revision.

  8. Expression of recombination-activating genes and T cell receptor gene recombination in the human T cell leukemia cell line

    Institute of Scientific and Technical Information of China (English)

    ZOU Hong-yun; MA Li; MENG Min-jie; YAO Xin-sheng; LIN Ying; WU Zhen-qiang; HE Xiao-wei; WANG Ju-fang; WANG Xiao-ning

    2007-01-01

    Background Recent studies have suggested that mature T cells can change their specificity through reexpression of recombination-activating genes (RAG) and RAG-mediated V(D)J recombination. This process is named receptor revision and has been observed in mature peripheral T cells from transgenic mice and human donors. However, whether the receptor revision in mature T cells is a random or orientated process remains poorly understood. Here we used the Jurkat human T cell line, which represents a mature stage of T cell development, as a model to investigate the regulation of T cell receptor (TCR) gene recombination.Methods TCR Dβ-Jβ signal joint T cell receptor excision DNA circles (sjTRECs) were determined by nested and seminested PCR. Double-strand DNA breaks at recombination signal sequences (RSSs) in the TCRVβ chain locus were detected by ligation-mediated-PCR. Further analysis of the complementarity-determining region 3 (CDR3) size of the TCRVβ chain was examined by the TCR GeneScan technique.Results RAG1, RAG2, and three crucial components of the nonhomologous DNA end-joining (NHEJ) pathway were readily detected in Jurkat. Characteristics of junctional diversity of Dβ2-Jβ2 signal joints and ds RSS breaks associated with the Dβ25' and Dβ 23' sites were detected in DNA from Jurkat cells. CDR3 size and the gene sequences of the TCRVβ chain did not change during cell proliferation.Conclusions RAG1 and RAG2 and ongoing TCR gene recombination are coexpressed in Jurkat cells, but the ongoing recombination process may not play a role in modification of the TCR repertoire. However, the results suggest that Jurkat could be used as a model for studying the regulation of RAGs and V(D)J recombination and as a "special" model of the coexistence of TCR gene rearrangements and "negative" receptor revision.

  9. Tools and Methods of Low-Frequency Radio Recombination Lines Investigations

    Science.gov (United States)

    Konovalenko, A. A.; Stepkin, S. V.; Mukha, D. V.; Vasilkovskiy, E. V.

    In the report the tools and methods of observations of radio recombination lines which are carried out at Institute of Radio Astronomy of the National Academy of Sciences of Ukraine using the world's largest decameter radio telescope UTR-2 (arrays "South - North" and "East - West") are described. The low-frequency radio recombination lines can be used as effective means of the low-density partially ionized interstellar medium diagnostic. However, low intensities of the lines and high level of interferences makes such investigations very difficult and impose high requirements to equipment. Observations are carried out with the 4096-channel digital correlometer and new generation digital spectral processors with 8192 spectral channels. Currently, the systematic observations of radio recombination lines have been carried out in the directions of remnants of supernova stars, Galactic plane, nebulas and dust clouds. Experiments aimed to finding the redshifted line of neutral hydrogen HI which arises in the cosmological epochs of reionization in the range 8 - 32 MHz are carried out. The carbon radio recombination lines have been detected in the direction of Cassiopeia A in the broad range of frequencies from 20 to 32 MHz. The carbon radio recombination line, corresponding to the transitions to atomic level with number of 1009 (these corresponds to the Bohr size of atom near 0,1 mm) have been registered.

  10. LOFAR observations of decameter carbon radio recombination lines towards Cassiopeia A

    Science.gov (United States)

    Salas, P.; Oonk, J. B. R.; van Weeren, R. J.; Salgado, F.; Morabito, L. K.; Toribio, M. C.; Emig, K.; Röttgering, H. J. A.; Tielens, A. G. G. M.

    2017-01-01

    We present a study of carbon radio recombination lines towards Cassiopeia A using LOFAR observations in the frequency range 10-33 MHz. Individual carbon α lines are detected in absorption against the continuum at frequencies as low as 16 MHz. Stacking several Cα lines we obtain detections in the 11-16 MHz range. These are the highest signal-to-noise measurements at these frequencies. The peak optical depth of the Cα lines changes considerably over the 11-33 MHz range with the peak optical depth decreasing from 4 × 10-3 at 33 MHz to 2 × 10-3 at 11 MHz, while the line width increases from 20 km s-1 to ˜150 km s-1. The combined change in peak optical depth and line width results in a roughly constant integrated optical depth. We interpret this as carbon atoms close to local thermodynamic equilibrium. In this work we focus on how the 11-33 MHz carbon radio recombination lines can be used to determine the gas physical conditions. We find that the ratio of the carbon radio recombination lines to that of the 158 μm [CII] fine-structure line is a good thermometer, while the ratio between low frequency carbon radio recombination lines provides a good barometer. By combining the temperature and pressure constraints with those derived from the line width we are able to constrain the gas properties (electron temperature and density) and radiation field intensity. Given the 1σ uncertainties in our measurements these are; Te ≈ 68-98 K, ne ≈ 0.02-0.035 cm-3 and Tr, 100 ≈ 1500-1650 K. Despite challenging RFI and ionospheric conditions, our work demonstrates that observations of carbon radio recombination lines in the 10-33 MHz range can provide insight into the gas conditions.

  11. USDA 846-1 fractal melon and derived recombinant inbred lines

    Science.gov (United States)

    The Agricultural Research Service, United States Department of Agriculture announces the release of a melon (Cucumis melo L.) breeding line with highly branched, fractal-type architectural growth habit and 81 derived recombinant inbred lines (RIL). The indeterminate, monoecious USDA 846-1 produces 2...

  12. Dynamics of male meiotic recombination frequency during plant development using Fluorescent Tagged Lines in Arabidopsis thaliana.

    Science.gov (United States)

    Li, Fan; De Storme, Nico; Geelen, Danny

    2017-02-13

    Meiotic homologous recombination plays a central role in creating genetic variability, making it an essential biological process relevant to evolution and crop breeding. In this study, we used pollen-specific fluorescent tagged lines (FTLs) to measure male meiotic recombination frequency during the development of Arabidopsis thaliana. Interestingly, a subset of pollen grains consistently shows loss of fluorescence expression in tested lines. Using nine independent FTL intervals, the spatio-temporal dynamics of male recombination frequency was assessed during plant development, considering both shoot type and plant age as independent parameters. In most genomic intervals assayed, male meiotic recombination frequency is highly consistent during plant development, showing no significant change between different shoot types and during plant aging. However, in some genomic regions, such as I1a and I5a, a small but significant effect of either developmental position or plant age were observed, indicating that the meiotic CO frequency in those intervals varies during plant development. Furthermore, from an overall view of all nine genomic intervals assayed, both primary and tertiary shoots show a similar dynamics of increasing recombination frequency during development, while secondary and lateral shoots remain highly stable. Our results provide new insights in the dynamics of male meiotic recombination frequency during plant development.

  13. Low Frequency Carbon Radio Recombination Lines I: Calculations of Departure Coefficients

    CERN Document Server

    Salgado, F; Oonk, J B R; Salas, P; Toribio, M C; Rottgering, H J A; Tielens, A G G M

    2016-01-01

    In the first paper of this series, we study the level population problem of recombining carbon ions. We focus our study on high quantum numbers anticipating observations of Carbon Radio Recombination Lines to be carried out by the LOw Frequency ARray (LOFAR). We solve the level population equation including angular momentum levels with updated collision rates up to high principal quantum numbers. We derive departure coefficients by solving the level population equation in the hydrogenic approximation and including low temperature dielectronic recombination effects. Our results in the hydrogenic approximation agree well with those of previous works. When comparing our results including dielectronic recombination we find differences which we ascribe to updates in the atomic physics (e.g., collision rates) and to the approximate solution method of the statistical equilibrium equations adopted in previous studies. A comparison with observations is discussed in an accompanying article, as radiative transfer effect...

  14. Development of Chromosomal Segment Substitution Lines from a Backcross Recombinant Inbred Population of Interspecific Rice Cross

    Institute of Scientific and Technical Information of China (English)

    CHEN Jie; Hafeez Ur Rahman BUGHIO; CHEN Da-zhou; LIU Guang-jie; ZHENG Kang-le; ZHUANG Jie-yun

    2006-01-01

    A backcross recombinant inbred line population consisting of 202 lines was developed from Xieqingzao B//Xieqingzao B Dongxiang wild rice. The population was assayed with DNA markers and phenotyped on planthopper resistance and yield traits. A linkage map consisting of 119 DNA markers and spanned for 1188 cM over the 12 rice chromosomes was constructed. Thirty-two chromosomal segment substitution lines were selected based on the percentage of Xieqingzao B allele at marker loci. These lines are of great potential for gene mapping and alien gene introgression.

  15. Physical conditions derived from OII recombination lines in planetary nebulae and their implications

    CERN Document Server

    Peimbert, Antonio; Delgado-Inglada, Gloria; Garcia-Rojas, Jorge; Peña, Miriam

    2014-01-01

    Based on high quality observations of multiplet V1 of OII and the NLTE atomic computations for OII we study the density and temperature of a sample of PNe. We find that, in general, the densities derived from recombination lines of OII are similar than the densities derived from forbidden lines. This implies that the signature for oxygen rich clumps of high density and low temperature is absent in most of the objects of our sample. Electron pressures derived from the hotter zones are similar or slightly larger than those derived from the colder zones, suggesting the presence of shock waves. The average temperatures and t2 values derived from H, He and O lines are similar and consistent with chemical homogeneity. These results suggest that the abundances of these objects are the ones derived from recombination lines.

  16. An efficient detection agent for the high throughput screening of recombinant manufacturing cell lines.

    Science.gov (United States)

    Duverger, Valérie; Sauvage, Christophe; Kobr, Michel; Imhof, Markus O

    2013-12-31

    To ensure the selection of high producing recombinant cell lines, a number of screening processes were developed in the presence of detection agents. Here, CHO cell lines secreting recombinant antibodies were detected in semi-solid medium containing detection agents. The aim was to compare two protein A-derived detection agents to two commercial fluorescent antibodies directed against the Fc part of the antibody of interest: the protein A derived Z domain fused to the red fluorescent protein and protein A labelled with a fluorescent Dylight™ 488 dye. All of these agents were compatible with cell recovery and colony formation, and specifically detected colonies secreting recombinant antibodies. Optimisation of the concentration of the fluorescent protein A allowed the identification of a higher number of good producers. Thus these data demonstrate that fluorescently labelled protein A-derivatives can be used for the selection of high producer cells.

  17. Genetic Analysis of Health-Related Secondary Metabolites in a Brassica rapa Recombinant Inbred Line Population

    NARCIS (Netherlands)

    Bagheri, H.; Soda, El M.; Kim, H.K.; Fritsche, S.; Jung, C.; Aarts, M.G.M.

    2013-01-01

    The genetic basis of the wide variation for nutritional traits in Brassica rapa is largely unknown. A new Recombinant Inbred Line (RIL) population was profiled using High Performance Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance (NMR) analysis to detect quantitative trait loci (QTLs) c

  18. Functional analysis of synthetic insectatachykinin analogs on recombinant neurokinin receptor expressing cell lines

    NARCIS (Netherlands)

    Torfs, H.; Akerman, K.E.; Nachman, R.J.; Oonk, H.B.; Detheux, M.; Poels, J.; Loy, van T.; Loof, A.; Meloen, R.H.; Vassart, G.; Parmentier, M.; Broeck, van den J.

    2002-01-01

    The activity of a series of synthetic tachykinin-like peptide analogs was studied by means of microscopic calcium imaging on recombinant neurokinin receptor expressing cell lines. A C-terminal pentapeptide (FTGMRa) is sufficient for activation of the stomoxytachykinin receptor (STKR) expressed in

  19. Optical Recombination Lines of Heavy-elements in Giant Extragalactic HII Regions

    CERN Document Server

    Esteban, C; Torres-Peimbert, S; Rodríguez, M

    2002-01-01

    We present high resolution observations of the giant extragalactic H II regions NGC 604, NGC 2363, NGC 5461 and NGC 5471, based on observations taken with the ISIS spectrograph on the William Herschel Telescope. We have detected -by the first time- C II and O II recombination lines in these objects. We find that recombination lines give larger C^{++} and O^{++} abundances than collisionallly excited lines, suggesting that temperature variations can be present in the objects. We detect [Fe IV] lines in NGC 2363 and NGC 5471, the most confident detection of optical lines of this kind in H II regions. Considering the temperature structure we derive their H, He, C, N, O, Ne, S, Ar, and Fe abundances. From the recombination lines of NGC 5461 and NGC 5471 we determine the presence of C/H and O/H gradients in M101. We calculate the Delta Y/Delta O and Delta Y/Delta Z values considering the presence of temperature variations and under the assumption of constant temperature. We obtain a better agreement with models of...

  20. Elemental abundances of Galactic bulge planetary nebulae from optical recombination lines

    CERN Document Server

    Wang, W

    2007-01-01

    (abridged) Deep long-slit optical spectrophotometric observations are presented for 25 Galactic bulge planetary nebulae (GBPNe) and 6 Galactic disk planetary nebulae (GDPNe). The spectra, combined with archival ultraviolet spectra obtained with the International Ultraviolet Explorer (IUE) and infrared spectra obtained with the Infrared Space Observatory (ISO), have been used to carry out a detailed plasma diagnostic and element abundance analysis utilizing both collisional excited lines (CELs) and optical recombination lines (ORLs). Comparisons of plasma diagnostic and abundance analysis results obtained from CELs and from ORLs reproduce many of the patterns previously found for GDPNe. In particular we show that the large discrepancies between electron temperatures (Te's) derived from CELs and from ORLs appear to be mainly caused by abnormally low values yielded by recombination lines and/or continua. Similarly, the large discrepancies between heavy element abundances deduced from ORLs and from CELs are large...

  1. Infrared recombination lines of hydrogen from young objects in the southern Galactic plane

    Science.gov (United States)

    Beck, Sara C.; Fischer, Jacqueline; Smith, Howard A.

    1991-01-01

    Near infrared recombination lines of hydrogen are observed in twelve young objects in the southern Galactic plane. The sample includes Herbig-Haro objects and IRAS dark-cloud point sources from the 1987 catalog of Persson and Campbell. In four of the IRAS sources two or three infrared lines are measured, and their intensity ratios are consistent with models of optically thick ionized winds. The intrinsic line shapes, retrieved from maximum-entropy deconvolutions, indicate gas velocities of 100 km/s or more as expected from ionized winds. These sources are apparently embedded pre-main-sequence objects with outflows. They include some of the brightest known YSOs.

  2. The HI/OH/Recombination line survey of the inner Milky Way (THOR)

    CERN Document Server

    Beuther, H; Rugel, M; Johnston, K; Wang, Y; Walter, F; Brunthaler, A; Walsh, A J; Ott, J; Stil, J; Henning, Th; Schierhuber, T; Kainulainen, J; Heyer, M; Goldsmith, P F; Anderson, L D; Longmore, S N; Klessem, R S; Glover, S C O; Urquhart, J S; Plume, R; Ragan, S E; Schneider, N; McClure-Griffiths, N M; Menten, K M; Smith, R; Roy, N; Shanahan, R; Nguyen-Luong, Q; Bigiel, F

    2016-01-01

    Context: The past decade has witnessed a large number of Galactic plane surveys at angular resolutions below 20". However, no comparable high-resolution survey exists at long radio wavelengths around 21cm in line and continuum emission. Methods: Employing the Very Large Array (VLA) in the C-array configuration and a large program, we observe the HI 21cm line, four OH lines, nineteen Halpha radio recombination lines as well as the continuum emission from 1 to 2GHz in full polarization over a large part of the first Galactic quadrant. Results: Covering Galactic longitudes from 14.5 to 67.4deg and latitudes between +-1.25deg, we image all of these lines and the continuum at ~20" resolution. These data allow us to study the various components of the interstellar medium (ISM): from the atomic phase, traced by the HI line, to the molecular phase, observed by the OH transitions, to the ionized medium, revealed by the cm continuum and the Halpha radio recombination lines. Furthermore, the polarized continuum emission...

  3. A suspended cell line from Trichoplusia ni (Lepidoptera):Characterization and expression of recombinant proteins

    Institute of Scientific and Technical Information of China (English)

    Min-Juan Meng; Tian-Long Li; Chang-You Li; Guo-Xun Li

    2008-01-01

    A suspended cell line from Trichoplusia ni embryos was established, and its susceptibility to Autographa californica multiple nuclear polyhedrosis virus (AcMNPV)infection was investigated. This cell line had characteristics distinct from the BTI-Tn5B 14 cell line (Tn5B 1-4) from T. ni in growth, and showed approximately the same responses to AcMNPV infection, production of occlusion bodies, and levels of recombinant protein expression. No clumps were observed at maximum cell density at late-log phase in shakeflask or T-flask cultures, and thus the cells represent a useful new contribution for baculovirus research. The cells consist of two major morphological types: approximately 70% spindle-shaped cells and 30% round cells. The cell line was highly susceptible to virus infection and produced around 107 AcMNPV occlusion bodies per cell, on average.Production of β-galactosidase and secreted alkaline phosphatase was high with 3.97 + 0.13×104 IU/mL and 3.48±0.40 IU/mL, respectively. This cell line may be applicable for studies of scale-up production of viruses or baculovirus-insect cell expression. We also believe the new line can be a source for cell clones with higher production of virus and recombinant proteins compared to the parent or other existing cell lines such as Tn5B 1-4.

  4. Recombination activating activity of XRCC1 analogous genes in X-ray sensitive and resistant CHO cell lines

    Science.gov (United States)

    Golubnitchaya-Labudová, O.; Portele, A.; Vaçata, V.; Lubec, G.; Rink, H.; Höfer, M.

    1997-10-01

    The XRCC1 gene (X-ray repair cross complementing) complements the DNA repair deficiency of the radiation sensitive Chinese hamster ovary (CHO) mutant cell line EM9 but the mechanism of the correction is not elucidated yet. XRCC1 shows substantial homology to the RAG2 gene (recombination activating gene) and we therefore tried to answer question, whether structural similarities (sequence of a putative recombination activating domain, aa 332-362 for XRCC1 and aa 286-316 in RAG2) would reflect similar functions of the homologous, putative recombination activating domain. PCR experiments revealed that no sequence homologous to the structural part of human XRCC1 was present in cDNA of CHO. Differential display demonstrated two putative recombination activating domains in the parental CHO line AA8 and one in the radiosensitive mutant EM9. Southern blot experiments showed the presence of several genes with partial homology to human XRCC1. Recombination studies consisted of expressing amplified target domains within chimeric proteins in recA - bacteria and subsequent detection of recombination events by sequencing the recombinant plasmids. Recombination experiments demonstrated recombination activating activity of all putative recombination activating domains amplified from AA8 and EM9 genomes as reflected by deletions within the insert of the recombinant plasmids. The recombination activating activity of XRCC1 analogues could explain a mechanism responsible for the correction of the DNA repair defect in EM9.

  5. Carbon and hydrogen radio recombination lines from the cold clouds towards Cassiopeia A

    Science.gov (United States)

    Oonk, J. B. R.; van Weeren, R. J.; Salas, P.; Salgado, F.; Morabito, L. K.; Toribio, M. C.; Tielens, A. G. G. M.; Röttgering, H. J. A.

    2017-02-01

    We use the Low Frequency Array to perform a systematic high spectral resolution investigation of the low-frequency 33-78 MHz spectrum along the line of sight to Cassiopeia A. We complement this with a 304-386 MHz Westerbork Synthesis Radio Telescope observation. In this first paper, we focus on the carbon radio recombination lines. We detect Cnα lines at -47 and -38 km s-1 in absorption for quantum numbers n = 438-584 and in emission for n = 257-278 with a high signal-to-noise ratio. These lines are associated with cold clouds in the Perseus spiral arm component. Hnα lines are detected in emission for n = 257-278. In addition, we also detect Cnα lines at 0 km s-1 associated with the Orion arm. We analyse the optical depth of these transitions and their linewidth. Our models show that the carbon line components in the Perseus arm are best fitted with an electron temperature of 85 K and an electron density of 0.04 cm-3 and can be constrained to within 15 per cent. The electron pressure is constrained to within 20 per cent. We argue that most of these carbon radio recombination lines arise in the CO-dark surface layers of molecular clouds, where most of the carbon is ionized, but hydrogen has made the transition from atomic to molecular. The hydrogen lines are clearly associated with the carbon line emitting clouds, but the low-frequency upper limits indicate that they likely do not trace the same gas. Combining the hydrogen and carbon results, we arrive at a firm lower limit to the cosmic-ray ionization rate of 2.5 × 10-18 s-1, but the actual value is likely much larger.

  6. Discovery of Carbon Radio Recombination Lines in absorption towards Cygnus~A

    CERN Document Server

    Oonk, J B R; Salgado, F; Morabito, L K; Tielens, A G G M; Rottgering, H J A; Asgekar, A; White, G J; Alexov, A; Anderson, J; Avruch, I M; Batejat, F; Beck, R; Bell, M E; van Bemmel, I; Bentum, M J; Bernardi, G; Best, P; Bonafede, A; Breitling, F; Brentjens, M; Broderick, J; Brueggen, M; Butcher, H R; Ciardi, B; Conway, J E; Corstanje, A; de Gasperin, F; de Geus, E; de Vos, M; Duscha, S; Eisloeffel, J; Engels, D; van Enst, J; Falcke, H; Fallows, R A; Fender, R; Ferrari, C; Frieswijk, W; Garrett, M A; Griessmeier, J; Hamaker, J P; Hassall, T E; Heald, G; Hessels, J W T; Hoeft, M; Horneffer, A; van der Horst, A; Iacobelli, M; Jackson, N J; Juette, E; Karastergiou, A; Klijn, W; Kohler, J; Kondratiev, V I; Kramer, M; Kuniyoshi, M; Kuper, G; van Leeuwen, J; Maat, P; Macario, G; Mann, G; Markoff, S; McKean, J P; Mevius, M; Miller-Jones, J C A; Mol, J D; Mulcahy, D D; Munk, H; Norden, M J; Orru, E; Paas, H; Pandey-Pommier, M; Pandey, V N; Pizzo, R; Polatidis, A G; Reich, W; Scaife, A M M; Schoenmakers, A; Schwarz, D; Shulevski, A; Sluman, J; Smirnov, O; Sobey, C; Stappers, B W; Steinmetz, M; Swinbank, J; Tagger, M; Tang, Y; Tasse, C; ter Veen, S; Thoudam, S; Toribio, C; van Nieuwpoort, R; Vermeulen, R; Vocks, C; Vogt, C; Wijers, R A M J; Wise, M W; Wucknitz, O; Yatawatta, S; Zarka, P; Zensus, A

    2014-01-01

    We present the first detection of carbon radio recombination line absorption along the line of sight to Cygnus A. The observations were carried out with the LOw Frequency ARray in the 33 to 57 MHz range. These low frequency radio observations provide us with a new line of sight to study the diffuse, neutral gas in our Galaxy. To our knowledge this is the first time that foreground Milky Way recombination line absorption has been observed against a bright extragalactic background source. By stacking 48 carbon $\\alpha$ lines in the observed frequency range we detect carbon absorption with a signal-to-noise ratio of about 5. The average carbon absorption has a peak optical depth of 2$\\times$10$^{-4}$, a line width of 10 km s$^{-1}$ and a velocity of +4 km s$^{-1}$ with respect to the local standard of rest. The associated gas is found to have an electron temperature $T_{e}\\sim$ 110 K and density $n_{e}\\sim$ 0.06 cm$^{-3}$. These properties imply that the observed carbon $\\alpha$ absorption likely arises in the c...

  7. Untangling the Recombination Line Emission from HII Regions with Multiple Velocity Components

    CERN Document Server

    Anderson, L D; Wenger, T V; Bania, T M; Balser, Dana S

    2015-01-01

    HII regions are the ionized spheres surrounding high-mass stars. They are ideal targets for tracing Galactic structure because they are predominantly found in spiral arms and have high luminosities at infrared and radio wavelengths. In the Green Bank Telescope HII Region Discovery Survey (GBT HRDS) we found that >30% of first Galactic quadrant HII regions have multiple hydrogen radio recombination line (RRL) velocities, which makes determining their Galactic locations and physical properties impossible. Here we make additional GBT RRL observations to determine the discrete HII region velocity for all 117 multiple-velocity sources within 18deg. < l < 65deg. The multiple-velocity sources are concentrated in the zone 22deg. < l < 32deg., coinciding with the largest regions of massive star formation, which implies that the diffuse emission is caused by leaked ionizing photons. We combine our observations with analyses of the electron temperature, molecular gas, and carbon recombination lines to determ...

  8. Australia telescope compact array observations of radio recombination lines toward 30 Doradus

    NARCIS (Netherlands)

    Peck, AB; Goss, WM; Dickel, HR; Roelfsema, PR; Kesteven, MJ; Dickel, [No Value; Milne, DK; Points, SD

    1997-01-01

    Three hydrogen recombination lines-H90 alpha at 8.9 GHz, H92 alpha at 8.3 GHz, and H109 alpha at 5.0 GHz-have been observed with the Australia Telescope Compact Array toward the 30 Doradus Nebula, the giant H II region in the Large Magellanic Cloud. In this paper, emphasis is placed on the more sens

  9. Low Frequency Carbon Radio Recombination Lines II: The Diffuse Interstellar Medium

    CERN Document Server

    Salgado, F; Oonk, J B R; Salas, P; Toribio, M C; Rottgering, H J A; Tielens, A G G M

    2016-01-01

    In the second paper of the series, we have modeled low frequency carbon radio recombination lines (CRRL) from the interstellar medium. Anticipating the LOw Frequency ARray (LOFAR) survey of Galactic CRRLs, we focus our study on the physical conditions of the diffuse cold neutral medium (CNM). We have used the improved departure coefficients computed in the first paper of the series to calculate line-to-continuum ratios. The results show that the line width and integrated optical depths of CRRL are sensitive probes of the electron density, gas temperature, and the emission measure of the cloud. Furthermore, the ratio of CRRL to the [CII] at 158 $\\mu$m line is a strong function of the temperature and density of diffuse clouds. Guided by our calculations, we analyze CRRL observations and illustrate their use with data from the literature.

  10. Nitrogen fertilization of coffee: organic compost and Crotalaria juncea L.

    Directory of Open Access Journals (Sweden)

    João Batista Silva Araujo

    2013-12-01

    Full Text Available Information concerning the response of coffee to organic fertilizers is scarce. This study evaluates the effect of different doses of compost and Crotalaria juncea L. on growth, production and nitrogen nutrition of coffee trees. The treatments consisted of compost at rates of 25, 50, 75 and 100% of the recommended fertilization, with or without the aerial part of C. juncea. C. juncea was grown with NH4-N (2% 15N and applied to coffee. The use of C. juncea increased growth in height and diameter of the coffee canopy. In the first year, the percentage of N derived from C. juncea reached 8.5% at seven months and 4.1% at fifteen months after fertilization. In the second year, the percentage of N derived from C. juncea reached 17.9% N at the early harvest, five months after fertilization. Increased rates of compost increased pH , P , K , Ca , Mg , sum of bases , effective CEC, base saturation and organic matter and reduced potential acidity. 15N allowed the identification of the N contribution from C. juncea with percentage of leaf N derived from Crotalaria juncea from 9.2 to 17.9%.

  11. Effective selection criteria for screening drought tolerant recombinant inbred lines of sunflower

    Directory of Open Access Journals (Sweden)

    Abdi Nishtman

    2013-01-01

    Full Text Available In this study, seventy two sunflower recombinant inbred lines were tested for their yielding ability under both water-stressed and well-watered states. The inbred lines were evaluated in a rectangular 8´9 lattice design with two replications in both well-watered and water-stressed conditions, separately. Eight drought tolerance indices including stability tolerance index (STI, mean productivity (MP, geometric mean productivity (GMP, harmonic mean (HM, stress susceptibility index (SSI, tolerance index (TOL, yield index (YI and yield stability index (YSI were calculated based on grain yield for every genotype. Results showed the highest values of mean productivity (MP index, geometric mean productivity (GMP, yield index (YI, harmonic mean (HM and stress tolerance index (STI indices for ‘C134a’ inbred line and least values of stress susceptibility index (SSI and tolerance (TOL for C61 inbred line. According to correlation of indices with yield performance under both drought stress and non-stress states and principle component analysis, indices including HM, MP, GMP and STI could properly distinguish drought tolerant sunflower inbred lines with high yield performance under both states. Cluster analysis of inbred lines using Ys, Yp and eight indices, categorized them into four groups including 19, 6, 26 and 19 inbred lines.

  12. Low-frequency Carbon Radio Recombination Lines. I. Calculations of Departure Coefficients

    Science.gov (United States)

    Salgado, F.; Morabito, L. K.; Oonk, J. B. R.; Salas, P.; Toribio, M. C.; Röttgering, H. J. A.; Tielens, A. G. G. M.

    2017-03-01

    In the first paper of this series, we study the level population problem of recombining carbon ions. We focus our study on high quantum numbers, anticipating observations of carbon radio recombination lines to be carried out by the Low Frequency Array. We solve the level population equation including angular momentum levels with updated collision rates up to high principal quantum numbers. We derive departure coefficients by solving the level population equation in the hydrogenic approximation and including low-temperature dielectronic capture effects. Our results in the hydrogenic approximation agree well with those of previous works. When comparing our results including dielectronic capture, we find differences that we ascribe to updates in the atomic physics (e.g., collision rates) and to the approximate solution method of the statistical equilibrium equations adopted in previous studies. A comparison with observations is discussed in an accompanying article, as radiative transfer effects need to be considered.

  13. Identification of two novel multiple recombinant avian leukosis viruses in two different lines of layer chicken.

    Science.gov (United States)

    Cai, Liming; Shen, Yanwei; Wang, Guihua; Guo, Huijun; Liu, Jianzhu; Cheng, Ziqiang

    2013-10-01

    Avian leukosis virus (ALV) is the most common oncogenetic retrovirus that emerges spontaneously as a result of recombination between exogenous viruses, exogenous viruses and endogenous viruses, and exogenous viruses and non-homologous cellular genes. In the present study, two natural recombinant avian leukosis viruses (rALVs) (LC110515-5 and LC110803-5) carrying a subgroup C gp85 gene, a subgroup E gp37 gene, and a subgroup J 3'UTR and 3'LTR were isolated from two different lines of layer flocks, Black-bone silky fowl (BSF) and commercial layer chicken, that suffered from myeloid leukosis. Although tumours were not observed in rALV-infected individual chickens, other non-neoplastic inflammatory lesions were evident. The two rALVs were cultured on DF-1 cells and identified by PCR, immunofluorescence assay and gene sequencing. The gp85 nucleotide sequence in the two isolates displayed a high identity (>95 %) with that of the gp85 gene in ALV-C, but the identity was less than 90 % with ALV-A/B/D/E and only 51 % with ALV-J. Phylogenetic analysis of the nucleotide and amino acid sequences confirmed that the two isolates were recombinant between ALV-C, ALV-E and ALV-J. Subgroup C ALV is rarely found in field cases. This report is the first to provide evidence that ALV-C has recombined with ALV-E and ALV-J in two different chicken lines. The source and characteristics of the two rALVs and ALV-C need to be further investigated.

  14. Diagnostics of recombining laser plasma parameters based on He-like ion resonance lines intensity ratios

    Science.gov (United States)

    Ryazantsev, S. N.; Skobelev, I. Yu; Faenov, A. Ya; Pikuz, T. A.; Grum-Grzhimailo, A. N.; Pikuz, S. A.

    2016-11-01

    While the plasma created by powerful laser expands from the target surface it becomes overcooled, i.e. recombining one. Improving of diagnostic methods applicable for such plasma is rather important problem in laboratory astrophysics nowadays because laser produced jets are fully scalable to young stellar objects. Such scaling is possible because of the plasma hydrodynamic equations invariance under some transformations. In this paper it is shown that relative intensities of the resonance transitions in He-like ions can be used to measure the parameters of recombining plasma. Intensity of the spectral lines corresponding to these transitions is sensitive to the density in the range of 1016-1020 cm-3 while the temperature ranges from 10 to 100 eV for ions with nuclear charge Zn ∼ 10. Calculations were carried out for F VIII ion and allowed to determine parameters of plasma jets created by nanosecond laser system ELFIE (Ecole Polytechnique, France) for astrophysical phenomenon modelling. Obtained dependencies are quite universal and can be used for any recombining plasma containing He-like fluorine ions.

  15. Implementation of Tidbinbilla 70-m On-The-Fly mapping and Hydrogen radio recombination line early results

    CERN Document Server

    Wong, G F; Green, J A; Tothill, N F H; Sugimoto, K; Filipovic, M D

    2016-01-01

    On-the-fly mapping of cm-wave spectral lines has been implemented at the the Tidbinbilla 70-m radio antenna. We describe the implementation and data reduction procedure and present new H92$\\alpha$ radio recombination line maps towards Orion A and Sagittarius A. Comparison of the Orion~A map to previous observations suggests that the lines arise largely from gas with electron density of 100--200\\,cm$^{-3}$. On-the-fly mapping is very efficient at generating large maps of bright lines (such as radio recombination lines), but will still yield strong efficiency gains for smaller maps of fainter lines, such as the ammonia inversion lines at the 1.3\\,cm wavelength.

  16. De novo transcriptome of Brassica juncea seed coat and identification of genes for the biosynthesis of flavonoids.

    Directory of Open Access Journals (Sweden)

    Xianjun Liu

    Full Text Available Brassica juncea, a worldwide cultivated crop plant, produces seeds of different colors. Seed pigmentation is due to the deposition in endothelial cells of proanthocyanidins (PAs, end products from a branch of flavonoid biosynthetic pathway. To elucidate the gene regulatory network of seed pigmentation in B. juncea, transcriptomes in seed coat of a yellow-seeded inbred line and its brown-seeded near- isogenic line were sequenced using the next-generation sequencing platform Illumina/Solexa and de novo assembled. Over 116 million high-quality reads were assembled into 69,605 unigenes, of which about 71.5% (49,758 unigenes were aligned to Nr protein database with a cut-off E-value of 10(-5. RPKM analysis showed that the brown-seeded testa up-regulated 802 unigenes and down-regulated 502 unigenes as compared to the yellow-seeded one. Biological pathway analysis revealed the involvement of forty six unigenes in flavonoid biosynthesis. The unigenes encoding dihydroflavonol reductase (DFR, leucoantho-cyanidin dioxygenase (LDOX and anthocyanidin reductase (ANR for late flavonoid biosynthesis were not expressed at all or at a very low level in the yellow-seeded testa, which implied that these genes for PAs biosynthesis be associated with seed color of B. juncea, as confirmed by qRT-PCR analysis of these genes. To our knowledge, it is the first time to sequence the transcriptome of seed coat in Brassica juncea. The unigene sequences obtained in this study will not only lay the foundations for insight into the molecular mechanisms underlying seed pigmentation in B.juncea, but also provide the basis for further genomics research on this species or its allies.

  17. CHO cell line specific prediction and control of recombinant monoclonal antibody N-glycosylation.

    Science.gov (United States)

    Grainger, Rhian K; James, David C

    2013-11-01

    Here we demonstrate that it is possible to predict and control N-glycan processing of a secreted recombinant monoclonal antibody during manufacturing process development using a combination of statistical modelling and comparative measurement of cell surface glycans using fluorescent lectins. Using design of experiments--response surface modelling (DoE-RSM) methodology to adjust the relative media concentrations of known metabolic effectors of galactosylation (manganese, galactose, and uridine) we have shown that β1,4-galactosylation of the same recombinant IgG4 monoclonal antibody produced by different CHO cell lines can be precisely controlled in a cell line specific manner. For two cell lines, monoclonal antibody galactosylation could be increased by over 100% compared to control, non-supplemented cultures without a reduction in product titre and with minimal effect on cell growth. Analysis of galactosylation effector interactions by DoE-RSM indicated that Mn²⁺ alone was necessary but not sufficient to improve galactosylation, and that synergistic combinations of Gal and Urd were necessary to maximize galactosylation, whilst minimizing the deleterious effect of Urd on cell growth. To facilitate rapid cell culture process development we also tested the hypothesis that substrate-level control of cellular galactosylation would similarly affect both cell surface and secreted monoclonal antibody glycans, enabling facile indirect prediction of product glycan processing. To support this hypothesis, comparative quantitation of CHO cell surface β1,4-galactosylation by flow cytometry using fluorescent derivatives of RCA and ConA lectins revealed that substrate-controlled variation in monoclonal antibody galactosylation and cell surface galactosylation were significantly correlated. Taken together, these data show that precision control of a complex, dynamic cellular process essential for the definition of protein product molecular heterogeneity and bioactivity is

  18. Generation of Cell Lines to Complement Adenovirus Vectors using Recombination-Mediated Cassette Exchange

    Directory of Open Access Journals (Sweden)

    Farley Daniel C

    2010-12-01

    Full Text Available Abstract Background Adenovirus serotype 5 (Ad5 has many favourable characteristics for development as a gene therapy vector. However, the utility of current Ad5 vectors is limited by transient transgene expression, toxicity and immunogenicity. The most promising form of vector is the high capacity type, which is deleted for all viral genes. However, these vectors can only be produced to relatively low titres and with the aid of helper virus. Therefore a continuing challenge is the generation of more effective Ad5 vectors that can still be grown to high titres. Our approach is to generate complementing cell lines to support the growth of Ad5 vectors with novel late gene deficiencies. Results We have used LoxP/Cre recombination mediated cassette exchange (RMCE to generate cell lines expressing Ad5 proteins encoded by the L4 region of the genome, the products of which play a pivotal role in the expression of Ad5 structural proteins. A panel of LoxP parent 293 cell lines was generated, each containing a GFP expression cassette under the control of a tetracycline-regulated promoter inserted at a random genome location; the cassette also contained a LoxP site between the promoter and GFP sequence. Clones displayed a variety of patterns of regulation, stability and level of GFP expression. Clone A1 was identified as a suitable parent for creation of inducible cell lines because of the tight inducibility and stability of its GFP expression. Using LoxP-targeted, Cre recombinase-mediated insertion of an L4 cassette to displace GFP from the regulated promoter in this parent clone, cell line A1-L4 was generated. This cell line expressed L4 100K, 22K and 33K proteins at levels sufficient to complement L4-33K mutant and L4-deleted viruses. Conclusions RMCE provides a method for rapid generation of Ad5 complementing cell lines from a pre-selected parental cell line, chosen for its desirable transgene expression characteristics. Parent cell lines can be

  19. Short-chain fluorescent tryptophan tags for on-line detection of functional recombinant proteins

    Directory of Open Access Journals (Sweden)

    Siepert Eva-Maria

    2012-09-01

    Full Text Available Abstract Background Conventional fluorescent proteins, such as GFP, its derivatives and flavin mononucleotide based fluorescent proteins (FbFPs are often used as fusion tags for detecting recombinant proteins during cultivation. These reporter tags are state-of-the-art; however, they have some drawbacks, which can make on-line monitoring challenging. It is discussed in the literature that the large molecular size of proteins of the GFP family may stress the host cell metabolism during production. In addition, fluorophore formation of GFP derivatives is oxygen-dependent resulting in a lag-time between expression and fluorescence detection and the maturation of the protein is suppressed under oxygen-limited conditions. On the contrary, FbFPs are also applicable in an oxygen-limited or even anaerobic environment but are still quite large (58% of the size of GFP. Results As an alternative to common fluorescent tags we developed five novel tags based on clustered tryptophan residues, called W-tags. They are only 5-11% of the size of GFP. Based on the property of tryptophan to fluoresce in absence of oxygen it is reasonable to assume that the functionality of our W-tags is also given under anaerobic conditions. We fused these W-tags to a recombinant protein model, the anti-CD30 receptor single-chain fragment variable antibody (scFv Ki-4(scFv and the anti-MucI single-chain fragment variable M12(scFv. During cultivation in Microtiter plates, the overall tryptophan fluorescence intensity of all cultures was measured on-line for monitoring product formation via the different W-tags. After correlation of the scattered light signal representing biomass concentration and tryptophan fluorescence for the uninduced cultures, the fluorescence originating from the biomass was subtracted from the overall tryptophan signal. The resulting signal, thus, represents the product fluorescence of the tagged and untagged antibody fragments. The product fluorescence signal

  20. Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase

    Directory of Open Access Journals (Sweden)

    Juliana Branco Novo

    2012-01-01

    Full Text Available Impaired activity of the lysosomal enzyme glucocerebrosidase (GCR results in the inherited metabolic disorder known as Gaucher disease. Current treatment consists of enzyme replacement therapy by administration of exogenous GCR. Although effective, it is exceptionally expensive, and patients worldwide have a limited access to this medicine. In Brazil, the public healthcare system provides the drug free of charge for all Gaucher’s patients, which reaches the order of $ 84 million per year. However, the production of GCR by public institutions in Brazil would reduce significantly the therapy costs. Here, we describe a robust protocol for the generation of a cell line producing recombinant human GCR. The protein was expressed in CHO-DXB11 (dhfr− cells after stable transfection and gene amplification with methotrexate. As expected, glycosylated GCR was detected by immunoblotting assay both as cell-associated (~64 and 59 kDa and secreted (63–69 kDa form. Analysis of subclones allowed the selection of stable CHO cells producing a secreted functional enzyme, with a calculated productivity of 5.14 pg/cell/day for the highest producer. Although being laborious, traditional methods of screening high-producing recombinant cells may represent a valuable alternative to generate expensive biopharmaceuticals in countries with limited resources.

  1. Generation of a Chinese Hamster Ovary Cell Line Producing Recombinant Human Glucocerebrosidase

    Science.gov (United States)

    Novo, Juliana Branco; Morganti, Ligia; Moro, Ana Maria; Paes Leme, Adriana Franco; Serrano, Solange Maria de Toledo; Raw, Isaias; Ho, Paulo Lee

    2012-01-01

    Impaired activity of the lysosomal enzyme glucocerebrosidase (GCR) results in the inherited metabolic disorder known as Gaucher disease. Current treatment consists of enzyme replacement therapy by administration of exogenous GCR. Although effective, it is exceptionally expensive, and patients worldwide have a limited access to this medicine. In Brazil, the public healthcare system provides the drug free of charge for all Gaucher's patients, which reaches the order of $ 84 million per year. However, the production of GCR by public institutions in Brazil would reduce significantly the therapy costs. Here, we describe a robust protocol for the generation of a cell line producing recombinant human GCR. The protein was expressed in CHO-DXB11 (dhfr−) cells after stable transfection and gene amplification with methotrexate. As expected, glycosylated GCR was detected by immunoblotting assay both as cell-associated (~64 and 59 kDa) and secreted (63–69 kDa) form. Analysis of subclones allowed the selection of stable CHO cells producing a secreted functional enzyme, with a calculated productivity of 5.14 pg/cell/day for the highest producer. Although being laborious, traditional methods of screening high-producing recombinant cells may represent a valuable alternative to generate expensive biopharmaceuticals in countries with limited resources. PMID:23091360

  2. LOFAR detections of low-frequency radio recombination lines towards Cassiopeia A

    CERN Document Server

    Asgekar, Ashish; Yatawatta, S; van Weeren, R J; McKean, J P; White, G; Jackson, N; Anderson, J; Avruch, I M; Batejat, F; Beck, R; Bell, M E; Bell, M R; van Bemmel, I; Bentum, M J; Bernardi, G; Best, P; Birzan, L; Bonafede, A; Braun, R; Breitling, F; van de Brink, R H; Broderick, J; Brouw, W N; Bruggen, M; Butcher, H R; van Cappellen, W; Ciardi, B; Conway, J E; de Gasperin, F; de Geus, E; de Jong, A; de Vos, M; Duscha, S; Eisloffel, J; Falcke, H; Fallows, R A; Ferrari, C; Frieswijk, W; Garrett, M A; Griesmeier, J-M; Grit, T; Gunst, A W; Hassall, T E; Heald, G; Hessels, J W T; Hoeft, M; Iacobelli, M; Intema, H; Juette, E; Karastergiou, A; Kohler, J; Kondratiev, V I; Kuniyoshi, M; Kuper, G; Law, C; van Leeuwen, J; Maat, P; Macario, G; Mann, G; Markoff, S; McKay-Bukowski, D; Mevius, M; Miller-Jones, J C A; Mol, J D; Morganti, R; Mulcahy, D D; Munk, H; Norden, M J; Orru, E; Paas, H; Pandey-Pommier, M; Pandey, V N; Pizzo, R; Polatidis, A G; Reich, W; Rottgering, H; Scheers, B; Schoenmakers, A; Sluman, J; Smirnov, O; Sobey, C; Steinmetz, M; Tagger, M; Tang, Y; Tasse, C; Vermeulen, R; Vocks, C; Wijers, R A M J; Wise, M W; Wucknitz, O; Zarka, P; 10.1051/0004-6361/201221001

    2013-01-01

    Cassiopeia A was observed using the Low-Band Antennas of the LOw Frequency ARray (LOFAR) with high spectral resolution. This allowed a search for radio recombination lines (RRLs) along the line-of-sight to this source. Five carbon-alpha RRLs were detected in absorption between 40 and 50 MHz with a signal-to-noise ratio of > 5 from two independent LOFAR datasets. The derived line velocities (v_LSR ~ -50 km/s) and integrated optical depths (~ 13 s^-1) of the RRLs in our spectra, extracted over the whole supernova remnant, are consistent within each LOFAR dataset and with those previously reported. For the first time, we are able to extract spectra against the brightest hotspot of the remnant at frequencies below 330 MHz. These spectra show significantly higher (15-80 %) integrated optical depths, indicating that there is small-scale angular structure on the order of ~1 pc in the absorbing gas distribution over the face of the remnant. We also place an upper limit of 3 x 10^-4 on the peak optical depths of hydro...

  3. Accelerating the inbreeding of multi-parental recombinant inbred lines generated by sibling matings.

    Science.gov (United States)

    Welsh, Catherine E; McMillan, Leonard

    2012-02-01

    Inbred model organisms are powerful tools for genetic studies because they provide reproducible genomes for use in mapping and genetic manipulation. Generating inbred lines via sibling matings, however, is a costly undertaking that requires many successive generations of breeding, during which time many lines fail. We evaluated several approaches for accelerating inbreeding, including the systematic use of back-crosses and marker-assisted breeder selection, which we contrasted with randomized sib-matings. Using simulations, we explored several alternative breeder-selection methods and monitored the gain and loss of genetic diversity, measured by the number of recombination-induced founder intervals, as a function of generation. For each approach we simulated 100,000 independent lines to estimate distributions of generations to achieve full-fixation as well as to achieve a mean heterozygosity level equal to 20 generations of randomized sib-mating. Our analyses suggest that the number of generations to fully inbred status can be substantially reduced with minimal impact on genetic diversity through combinations of parental backcrossing and marker-assisted inbreeding. Although simulations do not consider all confounding factors underlying the inbreeding process, such as a loss of fecundity, our models suggest many viable alternatives for accelerating the inbreeding process.

  4. VizieR Online Data Catalog: Water masers in M31. I. Recombination lines (Darling+, 2016)

    Science.gov (United States)

    Darling, J.; Gerard, B.; Amiri, N.; Lawrence, K.

    2016-09-01

    We constructed a catalog of 506 unresolved 24um sources from the Spitzer 24um map of M31 (Gordon et al. 2006ApJ...638L..87G); see Figure 1. Darling (2011ApJ...732L...2D) observed 206 24um sources in M31 using the Green Bank Telescope (GBT) in 2010 October through December. The 616-523 22.23508GHz ortho-water maser line observations were reported in Darling (2011ApJ...732L...2D), but simultaneous observations of the para-ammonia (NH3) rotational ground-state inversion transitions in the metastable states (J,K)=(1,1) and (2,2) at 23.6945 and 23.72263GHz, respectively, and the hydrogen recombination line H66α at 22.36417GHz were not. We subsequently observed all four of these lines toward an additional 300 24um sources in 2011 October through 2012 January. The resolution of the 24um Spitzer image is 6" (Gordon et al. 2006ApJ...638L..87G), so the unresolved IR sources remained within the 33" GBT beam even during the largest pointing drifts. The 33" beam (FWHM) at 22GHz spans 125pc in M31. (1 data file).

  5. Carbon and oxygen abundance gradients in NGC300 and M33 from optical recombination lines

    CERN Document Server

    Cipriano, L Toribio San; Esteban, C; Bresolin, F; Peimbert, M

    2016-01-01

    We present deep spectrophotometry of several Hii regions in the nearby low-mass spiral galaxies NGC 300 and M33. The data have been taken with UVES and OSIRIS spectrographs attached to the 8m VLT and 10.4m GTC telescopes, respectively. We have derived precise values of the physical conditions for each object making use of several emission line-intensity ratios. In particular, we have obtained direct determinations of the electron temperature in all the observed objects. We detect pure recombination lines (RLs) of Cii and Oii in several of the Hii regions, permitting to derive their C/H and C/O ratios. We have derived the radial abundance gradient of O for each galaxy making use of collisionally excited lines (CELs) and RLs, as well as the C and N gradients using RLs and CELs, respectively. We obtain the first determination of the C/H gradient of NGC 300 and improve its determination in the case of M33. In both galaxies, the C/H gradients are steeper that those of O/H, leading to negative C/O gradients. Compar...

  6. Modelling Carbon Radio Recombination Line observation towards the Ultra-Compact HII region W48A

    CERN Document Server

    Jeyakumar, S

    2013-01-01

    We model Carbon Recombination Line (CRL) emission from the Photo Dissociation Region (PDR) surrounding the Ultra-Compact (UC) HII region W48A. Our modelling shows that the inner regions ($A_V \\sim 1$) of the CII layer in the PDR contribute significantly to the CRL emission. The dependence of line ratios of CRL emission with the density of the PDR and the far ultra-violet (FUV) radiation incident on the region is explored over a large range of these parameters that are typical for the environments of UCHII regions. We find that by observing a suitable set of CRLs it is possible to constrain the density of the PDR. If the neutral density in the PDR is high ($\\gtrsim 10^7$ \\cmthree) CRL emission is bright at high frequencies ($\\gtrsim 20$ GHz), and absorption lines from such regions can be detected at low frequencies ($\\lesssim 10$ GHz). Modelling CRL observations towards W48A shows that the UCHII region is embedded in a molecular cloud of density of about $4 \\times$ 10$^7$ \\cmthree.

  7. VLA H53alpha radio recombination line observations of the ultraluminous infrared galaxy Arp~220

    CERN Document Server

    Rodriguez-Rico, C A; Viallefond, F; Zhao, J H; Gómez, Y; Anantharamaiah, K R

    2005-01-01

    We present high angular resolution (0.7'') observations made with the Very Large Array (VLA) of the radio recombination line (RRL) H53alpha and radio continuum emission at 43 GHz from the ultraluminous infrared galaxy (ULIRG) Arp 220. The 43 GHz continuum emission shows a compact structure (~2'') with two peaks separated by ~1'', the East (E) and West (W) components, that correspond to each galactic nucleus of the merger. The spectral indices for both the E and W components, using radio continuum images at 8.3 and 43 GHz are typical of synchrotron emission (alpha ~ -1.0). Our 43 GHz continuum and H53alpha line observations confirm the flux densities predicted by the models proposed by Anantharamaiah et al. This agreement with the models implies the presence of high-density (~ 100,000 cm^-3) compact HII regions (~ 0.1 pc) in Arp~220. The integrated H53alpha line emission is stronger toward the non-thermal radio continuum peaks, which are also coincident with the peaks of molecular emission of the H2CO. The coi...

  8. Stability of transgene expression, field performance and recombination breeding of transformed barley lines

    DEFF Research Database (Denmark)

    Horvath, H.; Jensen, L.G.; Wong, O.T.;

    2001-01-01

    originated from three independent primary transformants obtained by the biolistic method with three plasmids containing respectively, the bar gene, the uidA gene and the gene for a protein-engineered heat-stable (1,3-1,4)-beta -glucanase. Three production levels of recombinant beta -glucanase were identified...... in homozygous transgenic T-3 plants, and these remained constant over a 3-year period. In micro-malting experiments, the heat-stable enzyme reached levels of up to 1.4 mug.mg(-1) protein and survived kiln drying at levels of 70-100%. In the field trials of 1997 and 1998 the transgenic lines had a reduced 1000...

  9. Heavy elements in Galactic and Magellanic Cloud HII regions recombination-line versus forbidden-line abundances

    CERN Document Server

    Tsamis, Y G; Liu, X W; Danziger, I J; Storey, P J; Tsamis, Yiannis G.

    2003-01-01

    We have obtained deep optical, long-slit spectrophotometry of the Galactic HII regions M 17, NGC 3576 and of the Magellanic Cloud HII regions 30 Doradus, LMC N11B and SMC N66, recording the optical recombination lines (ORLs) of CII, NII and OII. Temperature-insensitive ORL C2+/O2+ and N2+/O2 ratios are obtained for all nebulae except SMC N66. The ORL C2+/O2+ ratios show remarkable agreement within each galactic system, while also being in agreement with the corresponding CEL ratios. For all five nebulae, the O2+/H+ abundance derived from multiple OII ORLs is found to be higher than the corresponding value derived from the strong [OIII] 4959, 5007A CELs, by factors of 1.8--2.7 for four of the nebulae. The LMC N11B nebula exhibits a more extreme discrepancy factor for the O2+ ion, ~5. Thus these HII regions exhibit ORL/CEL abundance discrepancy factors that are similar to those previously encountered amongst planetary nebulae. Our optical CEL O2+/H+ abundances agree to within 20-30 per cent with published O2+/H...

  10. Functional Coding Variation in Recombinant Inbred Mouse Lines Reveals Novel Serotonin Transporter-Associated Phenotypes

    Energy Technology Data Exchange (ETDEWEB)

    Carneiro, Ana [Vanderbilt University; Airey, David [University of Tennessee Health Science Center, Memphis; Thompson, Brent [Vanderbilt University; Zhu, C [Vanderbilt University; Rinchik, Eugene M [ORNL; Lu, Lu [University of Tennessee Health Science Center, Memphis; Chesler, Elissa J [ORNL; Erikson, Keith [University of North Carolina; Blakely, Randy [Vanderbilt University

    2009-01-01

    The human serotonin (5-hydroxytryptamine, 5-HT) transporter (hSERT, SLC6A4) figures prominently in the etiology or treatment of many prevalent neurobehavioral disorders including anxiety, alcoholism, depression, autism and obsessive-compulsive disorder (OCD). Here we utilize naturally occurring polymorphisms in recombinant inbred (RI) lines to identify novel phenotypes associated with altered SERT function. The widely used mouse strain C57BL/6J, harbors a SERT haplotype defined by two nonsynonymous coding variants (Gly39 and Lys152 (GK)). At these positions, many other mouse lines, including DBA/2J, encode Glu39 and Arg152 (ER haplotype), assignments found also in hSERT. Synaptosomal 5-HT transport studies revealed reduced uptake associated with the GK variant. Heterologous expression studies confirmed a reduced SERT turnover rate for the GK variant. Experimental and in silico approaches using RI lines (C57Bl/6J X DBA/2J=BXD) identifies multiple anatomical, biochemical and behavioral phenotypes specifically impacted by GK/ER variation. Among our findings are multiple traits associated with anxiety and alcohol consumption, as well as of the control of dopamine (DA) signaling. Further bioinformatic analysis of BXD phenotypes, combined with biochemical evaluation of SERT knockout mice, nominates SERT-dependent 5-HT signaling as a major determinant of midbrain iron homeostasis that, in turn, dictates ironregulated DA phenotypes. Our studies provide a novel example of the power of coordinated in vitro, in vivo and in silico approaches using murine RI lines to elucidate and quantify the system-level impact of gene variation.

  11. Quantitative Trait Loci Associated with Micronutrient Concentrations in Two Recombinant Inbred Wheat Lines

    Institute of Scientific and Technical Information of China (English)

    PU Zhi-en; WEI Yu-ming; ZHENG You-liang; YU Ma; HE Qiu-yi; CHEN Guo-yue; WANG Ji-rui; LIU Ya-xi; JIANG Qian-tao; LI Wei; DAI Shou-fen

    2014-01-01

    Micronutrient malnutrition affects over three billion people worldwide, especially women and children in developing countries. Increasing the bioavailable concentrations of essential elements in the edible portions of crops is an effective resolution to address this issue. To determine the genetic factors controlling micronutrient concentration in wheat, the quantitative trait locus (QTL) analysis for iron, zinc, copper, manganese, and selenium concentrations in two recombinant inbred line populations was performed. In all, 39 QTLs for ifve micronutrient concentrations were identiifed in this study. Of these, 22 alleles from synthetic wheat SHW-L1 and seven alleles from the progeny line of the synthetic wheat Chuanmai 42 showed an increase in micronutrient concentrations. Five QTLs on chromosomes 2A, 3D, 4D, and 5B found in both the populations showed signiifcant phenotypic variation for 2-3 micronutrient concentrations. Our results might help understand the genetic control of micronutrient concentration and allow the utilization of genetic resources of synthetic hexaploid wheat for improving micronutrient efifciency of cultivated wheat by using molecular marker-assisted selection.

  12. A high-yielding, generic fed-batch process for recombinant antibody production of GS-engineered cell lines

    DEFF Research Database (Denmark)

    Fan, Li; Zhao, Liang; Sun, Yating;

    2009-01-01

    An animal component-free and chemically defined fed-batch process for GS-engineered cell lines producing recombinant antibodies has been developed. The fed-batch process relied on supplying sufficient nutrients to match their consumption, simultaneously minimizing the accumulation of byproducts....... This generic and high-yielding fed-batch process would shorten development time, and ensure process stability, thereby facilitating the manufacture of therapeutic antibodies by GS-engineered cell lines....

  13. Phenotypic and molecular characterization of selected tomato recombinant inbred lines derived from the cross Solanum lycopersicum × S. pimpinellifolium

    Indian Academy of Sciences (India)

    Guillermo R. Pratta; Gustavo R. Rodriguez; Roxana Zorzoli; Estela M. Valle; Liliana A. Picardi

    2011-08-01

    An important trait defining fresh tomato marketability is fruit shelf life. Exotic germplasm of Solanum pimpinellifolium is able to prolong shelf life. Sixteen recombinant inbred lines with differing values of shelf life and fruit weight were derived by antagonistic-divergent selection from an interspecific cross involving Solanum pimpinellifolium. The objective of this study was to evaluate these recombinant inbred lines for many fruit quality traits such as diameter, height, size, acidity, colour, firmness, shelf life and weight, and to characterize them by amplified fragment length polymorphism markers. For most traits, a wide range of genetic variability was found and a wide range of molecular variation was also detected. Both sets of data allowed the identification of recombinant inbred lines by means of cluster analysis and principal component analysis. Genetic association among some amplified fragment length polymorphism markers and fruit quality traits, suggested by the principal component analysis, could be identified by single point analysis. Potential molecular markers underlying agronomical traits were detected in these recombinant inbred lines.

  14. Identification of QTLs underlying seed micronutrients accumulation in 'MD96-5722' by 'Spencer' recombinant inbred lines of soybean

    Science.gov (United States)

    Genetic mapping of quantitative trait loci (QTL) associated with seed nutrition levels is almost non-existent. The objective of this study was to identify QTLs associated with seed micronutrients accumulation (concentration) in a population of 92 F5:7 recombinant inbred lines (RILs) that derived fro...

  15. IDENTIFICATION OF A MAJOR QUANTITATIVE TRAIT LOCUS CONFERRING RICE BLAST RESISTANCE USING RECOMBINANT INBRED LINES

    Directory of Open Access Journals (Sweden)

    Sobrizal Sobrizal

    2013-05-01

    Full Text Available Blast disease caused by Pyricularia oryzae is one of the limiting factors for rice production world wide. The use of resistant varieties for managing blast disease is considered as the most eco-friendly approaches. However, their resistances may be broken down within a few years due to the appearance of new virulent blast races in the field. The objective of the present study was to identify the quantitative trait locus (QTL conferring resistance to blast disease using 126 recombinant inbred (RI lines originated from a crossing of a durably resistant upland rice genotype (Laka and a highly susceptible rice accession cultivar (Kencana Bali. The RI population was developed through a single seed descent method from 1997 to 2004. Resistance of the RI lines was evaluated for blast in an endemic area of Sukabumi, West Java, in 2005. Disease intensity of the blast was examined following the standard evaluation system developed by the International Rice Research Institute (IRRI. At the same year the RI lines were analyzed with 134 DNA markers. Results of the study showed that one major QTL was found to be associated with blast resistance, and this QTL was located near RM2136 marker on the long arm of chromosome 11. This QTL explained 87% of the phenotypic variation with 37% additive effect. The map position of this QTL differed from that of a partial resistant gene, Pi34, identified previously on chromosome 11 in the Japanese durably resistant variety, Chubu 32. The QTL, however, was almost at the same position as that of the multiple allele-resistant gene, Pik. Therefore, an allelic test should be conducted to clarify the allelic relationship between QTL identified in this study and the Pik. The RI lines are the permanent segregating population that could be very useful for analysing phenotypic variations of important agronomic traits possibly owned by the RI lines. The major QTL identified in this study could be used as a genetic resource in

  16. Phytoremediation of aspirin and tetracycline by Brassica juncea.

    Science.gov (United States)

    Gahlawat, Sonal; Gauba, Pammi

    2016-09-01

    With the increasing release of pharmaceutical drugs in the environment, research is in progress for investigating alternative methods for their remediation. Various studies have shown the phytoremediation potential of Brassica juncea for metals. The current study was aimed at evaluating the phytoremediation potential of B. juncea for two different pharmaceutical drugs i.e. aspirin and tetracycline in in-vitro conditions. The seeds of B. juncea were germinated and grown for a period of 28 and 24 days for aspirin and tetracycline, respectively. The study analyzed the remediation rate of B. juncea for the selected drugs in three different sets of varying concentration along with any phytotoxic effects exerted by the drugs on the seeds. Preliminary results showed that the average remediation rate of aspirin and tetracycline at the end of experiment was approximately 90% and 71%, respectively. As initial drug concentrations were increased in the media, the remediation rate also improved. However, at higher concentrations, the plants showed phytotoxicity as depicted by the decrease in shoot length of the germinated seeds. These preliminary results indicated that B. juncea could tolerate and remediate pharmaceutical drugs such as analgesics and antibiotics.

  17. Detection and mapping of QTL for earliness components in a bread wheat recombinant inbred lines population.

    Science.gov (United States)

    Hanocq, E; Niarquin, M; Heumez, E; Rousset, M; Le Gouis, J

    2004-12-01

    Earliness, an adaptative trait and factor of variation for agronomic characters, is a major trait in plant breeding. Its constituent traits, photoperiod sensitivity (PS), vernalization requirement (VR) and intrinsic earliness (IE), are largely under independent genetic controls. Mapping of major genes and quantitative trait loci (QTL) controlling these components is in progress. Most of the studies focusing on earliness considered it as a whole or through one (or two) of its components. The purpose of this study was to detect and map QTL for the three traits together through an experimental design combining field trials and controlled growth conditions. QTL were mapped in a population of F(7) recombinant inbred lines derived by single-seed descent from a cross between two French varieties, 'Renan' and 'Recital'. A map was previously constructed, based on 194 lines and 254 markers, covering about 77% of the genome. Globally, 13 QTL with a LOD>2.5 were detected, of which four control PS, five control VR and four control IE. Two major photoperiod sensitive QTL, together explaining more than 31% of the phenotypic variation, were mapped on chromosomes 2B and 2D, at the same position as the two major genes Ppd-B1 and Ppd-D1. One major VR QTL explaining (depending on the year) 21.8-39.6% of the phenotypic variation was mapped on 5A. Among the other QTL, two QTL of PS and VR not referenced so far were detected on 5A and 6D, respectively. A VR QTL already detected on 2B in a connected population was confirmed.

  18. Phenotypic and genotypic variation among Capsicum annuum recombinant inbred lines resistant to bacterial spot.

    Science.gov (United States)

    Moreira, S O; Rodrigues, R; Oliveira, H S; Medeiros, A M; Sudré, C P; Gonçalves, L S A

    2013-04-17

    A breeding program carried out under Brazilian growing conditions to obtain Capsicum annuum cultivars with disease resistance to bacterial spot (BS) produced 8 promising recombinant inbred lines (RILs). The present study aimed to characterize these RILs using phenotypic descriptors and molecular markers (inter-simple sequence repeat) and to confirm their resistance to BS. Twenty-two phenotypic descriptors and 15 inter-simple sequence repeat primers were used to characterize the RILs. The parent, UENF 1381, which is resistant to BS, and 'Casca Dura Ikeda', a traditional cultivar, were used as standards. Variability among genotypes was observed considering either binary or multicategorical characteristics, such as fruit length, fruit diameter, and fruit longitudinal and transversal section. Such variability in fruit traits can be exploited to develop new genotypes with BS resistance for various types of market consumption. RILs numbered 1, 3, and 6 were the most homogenous, whereas those coded 2, 5, 8, and 11 had the same level of heterogeneity as that observed in 'Casca Dura Ikeda'. Molecular analysis clustered the genotypes into 5 groups, with RILs 1, 2, 3, and 5 allocated in isolated groups. RILs 1, 2, 6, and 8 confirmed resistance to BS. Considering homogeneity level and BS resistance, RILs 1 and 6 were suitable for use as pre-cultivars in final tests to register and release two new C. annuum cultivars.

  19. Genomic dissection of drought resistance in durum wheat x wild emmer wheat recombinant inbreed line population.

    Science.gov (United States)

    Peleg, Zvi; Fahima, Tzion; Krugman, Tamar; Abbo, Shahal; Yakir, Dan; Korol, Abraham B; Saranga, Yehoshua

    2009-07-01

    Drought is the major factor limiting wheat productivity worldwide. The gene pool of wild emmer wheat, Triticum turgidum ssp. dicoccoides, harbours a rich allelic repertoire for morpho-physiological traits conferring drought resistance. The genetic and physiological bases of drought responses were studied here in a tetraploid wheat population of 152 recombinant inbreed lines (RILs), derived from a cross between durum wheat (cv. Langdon) and wild emmer (acc# G18-16), under contrasting water availabilities. Wide genetic variation was found among RILs for all studied traits. A total of 110 quantitative trait loci (QTLs) were mapped for 11 traits, with LOD score range of 3.0-35.4. Several QTLs showed environmental specificity, accounting for productivity and related traits under water-limited (20 QTLs) or well-watered conditions (15 QTLs), and in terms of drought susceptibility index (22 QTLs). Major genomic regions controlling productivity and related traits were identified on chromosomes 2B, 4A, 5A and 7B. QTLs for productivity were associated with QTLs for drought-adaptive traits, suggesting the involvement of several strategies in wheat adaptation to drought stress. Fifteen pairs of QTLs for the same trait were mapped to seemingly homoeologous positions, reflecting synteny between the A and B genomes. The identified QTLs may facilitate the use of wild alleles for improvement of drought resistance in elite wheat cultivars.

  20. Genetic Analysis of Health-Related Secondary Metabolites in a Brassica rapa Recombinant Inbred Line Population

    Directory of Open Access Journals (Sweden)

    Mark G. M. Aarts

    2013-07-01

    Full Text Available The genetic basis of the wide variation for nutritional traits in Brassica rapa is largely unknown. A new Recombinant Inbred Line (RIL population was profiled using High Performance Liquid Chromatography (HPLC and Nuclear Magnetic Resonance (NMR analysis to detect quantitative trait loci (QTLs controlling seed tocopherol and seedling metabolite concentrations. RIL population parent L58 had a higher level of glucosinolates and phenylpropanoids, whereas levels of sucrose, glucose and glutamate were higher in the other RIL population parent, R-o-18. QTL related to seed tocopherol (α-, β-, γ-, δ-, α-⁄γ- and total tocopherol concentrations were detected on chromosomes A3, A6, A9 and A10, explaining 11%–35% of the respective variation. The locus on A3 co-locates with the BrVTE1gene, encoding tocopherol cyclase. NMR spectroscopy identified the presence of organic/amino acid, sugar/glucosinolate and aromatic compounds in seedlings. QTL positions were obtained for most of the identified compounds. Compared to previous studies, novel loci were found for glucosinolate concentrations. This work can be used to design markers for marker-assisted selection of nutritional compounds in B. rapa.

  1. The Green Bank Telescope H II Region Discovery Survey: IV. Helium and Carbon Recombination Lines

    CERN Document Server

    Wenger, Trey V; Balser, Dana S; Anderson, L D

    2012-01-01

    The Green Bank Telescope H II Region Discovery Survey (GBT HRDS) found hundreds of previously unknown Galactic regions of massive star formation by detecting hydrogen radio recombination line (RRL) emission from candidate H II region targets. Since the HRDS nebulae lie at large distances from the Sun, they are located in previously unprobed zones of the Galactic disk. Here we derive the properties of helium and carbon RRL emission from HRDS nebulae. Our target sample is the subset of the HRDS that has visible helium or carbon RRLs. This criterion gives a total of 84 velocity components (14% of the HRDS) with helium emission and 52 (9%) with carbon emission. For our highest quality sources, the average ionic He-4+/H+ abundance ratio by number, , is 0.068 +/- 0.023 (1-sigma). This is the same ratio as that measured for the sample of previously known Galactic H II regions. Nebulae without detected helium emission give robust y+ upper limits. There are 5 RRL emission components with y+ less than 0.04 and another ...

  2. THE GREEN BANK TELESCOPE H II REGION DISCOVERY SURVEY. IV. HELIUM AND CARBON RECOMBINATION LINES

    Energy Technology Data Exchange (ETDEWEB)

    Wenger, Trey V.; Bania, T. M. [Astronomy Department, 725 Commonwealth Avenue, Boston University, Boston, MA 02215 (United States); Balser, Dana S. [National Radio Astronomy Observatory, 520 Edgemont Road, Charlottesville, VA, 22903-2475 (United States); Anderson, L. D. [Department of Physics, West Virginia University, Morgantown, WV 26506 (United States)

    2013-02-10

    The Green Bank Telescope H II Region Discovery Survey (GBT HRDS) found hundreds of previously unknown Galactic regions of massive star formation by detecting hydrogen radio recombination line (RRL) emission from candidate H II region targets. Since the HRDS nebulae lie at large distances from the Sun, they are located in previously unprobed zones of the Galactic disk. Here, we derive the properties of helium and carbon RRL emission from HRDS nebulae. Our target sample is the subset of the HRDS that has visible helium or carbon RRLs. This criterion gives a total of 84 velocity components (14% of the HRDS) with helium emission and 52 (9%) with carbon emission. For our highest quality sources, the average {sup 4}He{sup +}/H{sup +} abundance ratio by number, (y {sup +}), is 0.068 {+-} 0.023(1{sigma}). This is the same ratio as that measured for the sample of previously known Galactic H II regions. Nebulae without detected helium emission give robust y {sup +} upper limits. There are 5 RRL emission components with y {sup +} less than 0.04 and another 12 with upper limits below this value. These H II regions must have either a very low {sup 4}He abundance or contain a significant amount of neutral helium. The HRDS has 20 nebulae with carbon RRL emission but no helium emission at its sensitivity level. There is no correlation between the carbon RRL parameters and the 8 {mu}m mid-infrared morphology of these nebulae.

  3. Carbon and oxygen abundance gradients in NGC 300 and M33 from optical recombination lines

    Science.gov (United States)

    Toribio San Cipriano, L.; García-Rojas, J.; Esteban, C.; Bresolin, F.; Peimbert, M.

    2016-05-01

    We present deep spectrophotometry of several H II regions in the nearby low-mass spiral galaxies NGC 300 and M33. The data have been taken with Ultraviolet-Visual Echelle Spectrograph and Optical System for Imaging and low-Intermediate-Resolution Integrated Spectroscopy spectrographs attached to the 8-m Very Large Telescope and 10.4-m Gran Telescopio Canarias telescopes, respectively. We have derived precise values of the physical conditions for each object making use of several emission line-intensity ratios. In particular, we have obtained direct determinations of the electron temperature in all the observed objects. We detect pure recombination lines (RLs) of C II and O II in several of the H II regions, permitting to derive their C/H and C/O ratios. We have derived the radial abundance gradient of O for each galaxy making use of collisionally excited lines (CELs) and RLs, as well as the C and N gradients using RLs and CELs, respectively. We obtain the first determination of the C/H gradient of NGC 300 and improve its determination in the case of M33. In both galaxies, the C/H gradients are steeper than those of O/H, leading to negative C/O gradients. Comparing with similar results for other spiral galaxies, we find a strong correlation between the slope of the C/H gradient and MV. We find that some H II regions located close to the isophotal radius (R25) of NGC 300 and M33 show C/O ratios more similar to those typical of dwarf galaxies than those of H II regions in the discs of more massive spirals. This may be related to the absence of flattening of the gradients in the external parts of NGC 300 and M33. Finally, we find very similar N/H gradients in both galaxies and a fair correlation between the slope of the N/H gradient and MV comparing with similar data for a sample of spiral galaxies.

  4. Accumulation of Gold Nanoparticles in Brassic Juncea

    Energy Technology Data Exchange (ETDEWEB)

    Marshall, A.T.; Haverkamp, R.G.; Davies, C.E.; Parsons, J.G.; Gardea-Torresdey, J.L.; Agterveld, D.van

    2009-06-03

    Enzymatic digestion is proposed as a method for concentrating gold nanoparticles produced in plants. The mild conditions of digestion are used in order to avoid an increase in the gold particle size, which would occur with a high-temperature process, so that material suitable for catalysis may be produced. Gold nanoparticles of a 5-50-nm diameter, as revealed by transmission electron microscopy (TEM), at concentrations 760 and 1120 ppm Au, were produced within Brassica juncea grown on soil with 22-48 mg Au kg{sup -1}. X-ray absorption near edge spectroscopy (XANES) reveals that the plant contained approximately equal quantities of Au in the metallic (Au{sup 0}) and oxidized (Au{sup +1}) states. Enzymatic digestion dissolved 55-60 wt% of the plant matter. Due to the loss of the soluble gold fraction, no significant increase in the total concentration of gold in the samples was observed. However, it is likely that the concentration of the gold nanoparticles increased by a factor of two. To obtain a gold concentration suitable for catalytic reactions, around 95 wt% of the starting dry biomass would need to be solubilized or removed, which has not yet been achieved.

  5. Accumulation of gold nanoparticles in Brassic juncea.

    Science.gov (United States)

    Marshall, Aaron T; Haverkamp, Richard G; Davies, Clive E; Parsons, Jason G; Gardea-Torresdey, Jorge L; van Agterveld, Dimitri

    2007-01-01

    Enzymatic digestion is proposed as a method for concentrating gold nanoparticles produced in plants. The mild conditions of digestion are used in order to avoid an increase in the gold particle size, which would occur with a high-temperature process, so that material suitable for catalysis may be produced. Gold nanoparticles of a 5-50-nm diameter, as revealed by transmission electron microscopy (TEM), at concentrations 760 and 1120 ppm Au, were produced within Brassica juncea grown on soil with 22-48 mg Au kg(-1). X-ray absorption near edge spectroscopy (XANES) reveals that the plant contained approximately equal quantities of Au in the metallic (Au0) and oxidized (Au+1) states. Enzymatic digestion dissolved 55-60 wt% of the plant matter. Due to the loss of the soluble gold fraction, no significant increase in the total concentration of gold in the samples was observed. However, it is likely that the concentration of the gold nanoparticles increased by a factor of two. To obtain a gold concentration suitable for catalytic reactions, around 95 wt% of the starting dry biomass would need to be solubilized or removed, which has not yet been achieved.

  6. Microcarrier culture of lepidopteran cell lines: implications for growth and recombinant protein production.

    Science.gov (United States)

    Ikonomou, Laertis; Drugmand, Jean-Christophe; Bastin, Georges; Schneider, Yves-Jacques; Agathos, Spiros N

    2002-01-01

    Several microcarrier systems were screened with Sf-9 and High-Five cell lines as to their ability to support cell growth and recombinant (beta-galactosidase) protein production. Growth of both cell lines on compact microcarriers, such as Cytodex-1 and glass beads, was minimal, as cells detached easily from the microcarrier surface and grew as single cells in the medium. Cell growth was also problematic on Cytopore-1 and -2 porous microcarriers. Cells remained attached for several days inside the microcarrier pores, but no cell division and proliferation were observed. On the contrary, insect cells grew well in the interior of Fibra-Cel disks mainly as aggregates at points of fiber intersection, reaching final (plateau) densities of about 4 x 10(6) (Sf-9) and 2.7 x 10(6) (High-Five) cells mL(-1) (8 x 10(6) and 5.5 x 10(6) cells per cm(2) of projected disk area, respectively). Their growth was described well by the logistic equation, which takes into account possible inhibition effects. Beta-Galactosidase (beta-gal) production of Sf-9 cells on Fibra-Cel disks (infected at 3.3 x 10(6) cells mL(-1)) was prolonged (192 h), and specific protein production was similar to that of high-density free cell infection. Cultispher-S microcarriers were found to be a very efficient system for the growth of High-Five cells, whereas no growth of Sf-9 cells took place for the same system. Concentrations of about 9 x 10(6) cells mL(-1) were reached within 120 h, with cell growth in both microcarriers and aggregates, appearance of cellular bridges between microcarriers and aggregates, and eventual formation of macroaggregates incorporating several microcarriers. Specific protein productions after beta-gal baculovirus infection at increasing cell concentrations were almost constant, thus leading to elevated volumetric protein production: final beta-gal titers of 946, 1728, and 1484 U mL(-1) were obtained for infection densities of 3.4, 7.2, and 8.9 x 10(6) cells mL(-1), respectively.

  7. Allelic effects on starch structure and properties of six starch biosynthetic genes in a rice recombinant inbred line population

    OpenAIRE

    2015-01-01

    Background The genetic diversity of six starch biosynthetic genes (Wx, SSI, SSIIa, SBEI, SBEIIa and SBEIIb) in indica and japonica rices opens an opportunity to produce a new variety with more favourable grain starch quality. However, there is limited information about the effects of these six gene allele combinations on starch structure and properties. A recombinant inbred line population from a cross between indica and japonica varieties offers opportunities to combine specific alleles of t...

  8. Identification for Heat Tolerance in Backcross Recombinant Lines and Screening of Backcross Introgression Lines with Heat Tolerance at Milky Stage in Rice

    Institute of Scientific and Technical Information of China (English)

    LIAO Jiang-lin; ZHANG Hong-yu; SHAO Xue-lian; ZHONG Ping-an; HUANG Ying-jin

    2011-01-01

    The present study aimed at breeding new rice germplasms with similar genome but significantly differed in heat tolerance during the grain filling stage.A total of 791 BC1F8 backcross recombinant lines,derived from the cross of Xieqingzao B / N22∥Xieqingzao B,were used as materials.Each rice line was separated evenly into two groups,and the heat tolerance of all rice lines were evaluated at natural high temperature in fields.The rice lines with heat tolerant index higher than 90% or lower than 40% were selected to compare the phenotypic characters and further identify heat tolerance at the early milky stage in a phytotron.Rice lines with similar phenotypic characters but significantly differed in heat tolerance at the milky stage were analyzed by 887 simple sequence repeat markers that were evenly distributed on the 12 rice chromosomes.In the result,12 (6 pairs) rice lines with similar phenotypic characters but significantly differed in heat tolerance at the milky stage were obtained.Molecular marker analysis indicated that the genomic polymorphism between 703T and 704S was the smallest in the 6 pairs of rice lines,with only 16 polymorphic sites,including 22 different alleles.The application of these two backcross introgression rice lines for future study on the mechanisms of heat tolerance in rice at the milky stage will be theoretically beneficial in reducing the interference caused by genetic differences from experimental materials.

  9. A deep survey of heavy element lines in planetary nebulae -- II. Recombination line abundances and evidence for ultra-cold plasma

    CERN Document Server

    Tsamis, Y G; Liu, X W; Storey, P J; Danziger, I J

    2004-01-01

    [Abridged] Deep optical observations of the spectra of 12 Galactic planetary nebulae (PNe) and 3 Magellanic Cloud PNe were presented in Paper I by Tsamis et al. (2003b), who carried out an abundance analysis using the collisionally excited forbidden lines. Here, the relative intensities of faint optical recombination lines (ORLs) from ions of carbon, nitrogen and oxygen are analysed in order to derive the abundances of these ions relative to hydrogen. We define an abundance discrepancy factor (ADF) as the ratio of the abundance derived for a heavy element ion from its recombination lines to that derived for the same ion from its ultraviolet, optical or infrared collisionally excited lines (CELs). All of the PNe in our sample are found to have ADF's that exceed unity. There is no dependence of the magnitude of the ADF upon the excitation energy of the UV, optical or IR CEL transition used, indicating that classical nebular temperature fluctuations--i.e. in a chemically homogeneous medium--are not the cause of ...

  10. Increasing the stearate content in seed oil of Brassica juncea by heterologous expression of MlFatB affects lipid content and germination frequency of transgenic seeds.

    Science.gov (United States)

    Bhattacharya, Surajit; Sinha, Saheli; Das, Natasha; Maiti, Mrinal K

    2015-11-01

    Fatty acids from dietary lipids can impart both beneficial and harmful health effects. The compositional balance between saturated and unsaturated fatty acids plays a decisive role in maintaining the physiological harmony, proper growth and development in the human system. In case of Brassica juncea seed oil, the level of saturated fatty acid, especially desirable stearate is very much lower than the recommended value, along with a high content of nutritionally undesirable erucic acid. Therefore, in order to shift the carbon flux towards the production of stearate at the expense of erucate, the MlFatB gene encoding a FatB thioesterase from Madhuca longifolia (latifolia) was expressed heterologously in seed tissues of B. juncea. The functional MlFatB competed with the highly active endogenous BjFatA thioesterase, and the transgenic B. juncea lines showed noteworthy changes in their seed fatty acid profiles. The proportion of stearate increased up to 16-fold, constituting almost 31% of the total fatty acids along with the production of arachidic acid in significant amount (up to ∼11%). Moreover, the content of erucate was reduced up to 71% in the seed oils of transgenic lines. Although a nutritionally desirable fatty acid profile was achieved, the transgenic seeds exhibit reduction or abolition of seed germination in addition to a decrease in seed lipid content. The findings of the present study revealing the stearoyl-ACP thioesterase-mediated enhancement of the stearate content that is associated with reduced germination frequency of transgenic B. juncea seeds, may explain why no natural or induced stearate-rich Brassica has been found or developed. Furthermore, this study also suggests that the newly characterized MlFatB is a potential candidate gene for refined metabolic engineering strategy in B. juncea or other plant species for increasing stearate content in seed oil.

  11. The HI/OH/Recombination line survey of the inner Milky Way (THOR). Survey overview and data release 1

    Science.gov (United States)

    Beuther, H.; Bihr, S.; Rugel, M.; Johnston, K.; Wang, Y.; Walter, F.; Brunthaler, A.; Walsh, A. J.; Ott, J.; Stil, J.; Henning, Th.; Schierhuber, T.; Kainulainen, J.; Heyer, M.; Goldsmith, P. F.; Anderson, L. D.; Longmore, S. N.; Klessen, R. S.; Glover, S. C. O.; Urquhart, J. S.; Plume, R.; Ragan, S. E.; Schneider, N.; McClure-Griffiths, N. M.; Menten, K. M.; Smith, R.; Roy, N.; Shanahan, R.; Nguyen-Luong, Q.; Bigiel, F.

    2016-10-01

    Context. The past decade has witnessed a large number of Galactic plane surveys at angular resolutions below 20''. However, no comparable high-resolution survey exists at long radio wavelengths around 21 cm in line and continuum emission. Aims: We remedy this situation by studying the northern Galactic plane at 20'' resolution in emission of atomic, molecular, and ionized gas. Methods: Employing the Karl G. Jansky Very Large Array (VLA) in the C-array configuration and a large program, we observe the HI 21 cm line, four OH lines, nineteen Hnα radio recombination lines as well as the continuum emission from 1 to 2 GHz in full polarization over a large part of the first Galactic quadrant. Results: Covering Galactic longitudes from 14.5 to 67.4 deg and latitudes between ± 1.25 deg, we image all of these lines and the continuum at 20'' resolution. These data allow us to study the various components of the interstellar medium (ISM): from the atomic phase, traced by the HI line, to the molecular phase, observed by the OH transitions, to the ionized medium, revealed by the cm continuum and the Hnα radio recombination lines. Furthermore, the polarized continuum emission enables magnetic field studies. In this overview paper, we discuss the survey outline and present the first data release as well as early results from the different datasets. We now release the first half of the survey; the second half will follow later after the ongoing data processing has been completed. The data in fits format (continuum images and line data cubes) can be accessed through the project web-page. Conclusions: The HI/OH/Recombination line survey of the Milky Way (THOR) opens a new window to the different parts of the ISM. It enables detailed studies of molecular cloud formation, conversion of atomic to molecular gas, and feedback from Hii regions as well as the magnetic field in the Milky Way. It is highly complementary to other surveys of our Galaxy, and comparing the different datasets

  12. Analysis of recombinant Schistosoma mansoni antigen rSmp28 by on-line liquid chromatography-mass spectrometry combined with sodium dodecyl sulfate polyacrylamide gel electrophoresis

    NARCIS (Netherlands)

    Klarskov, K.; Roecklin, D.; Bouchon, B.; Sabatie, J.; Van Dorsselaer, A.; Bischoff, Rainer

    1994-01-01

    A recombinant Schistosoma mansoni antigen produced in Saccharomyces cerevisiae and purified by glutathione-Sepharose affinity chromatography was analyzed by tryptic peptide mapping using on-line reversed-phase high-performance liquid chromatography pneumatically assisted electrospray mass

  13. ALMA observations of the submillimetre hydrogen recombination line from the type 2 active nucleus of NGC 1068

    CERN Document Server

    Izumi, Takuma; Imanishi, Masatoshi; Kohno, Kotaro

    2016-01-01

    Hydrogen recombination lines at the submillimetre band (submm-RLs) can serve as probes of ionized gas without dust extinction. One therefore expects to probe the broad line region (BLR) of an obscured (type 2) active galactic nucleus (AGN) with those lines. However, admitting the large uncertainty in the continuum level, here we report on the non-detection of both broad and narrow H26$\\alpha$ emission line (rest frequency = 353.62 GHz) towards the prototypical type 2 AGN of NGC 1068 with the Atacama Large Millimeter/submillimeter Array (ALMA). We also investigate the nature of BLR clouds that can emit submm-RLs with model calculations. As a result, we suggest that clouds with an electron density ($N_e$) of $\\sim$ 10$^9$ cm$^{-3}$ can mainly contribute to broad submm-RLs in terms of the line flux. On the other hand, line flux from other density clouds would be insignificant considering their too large or too small line optical depths. Since $N_e$ $\\sim$ 10$^9$ cm$^{-3}$ is typical in BLRs, insufficient sensiti...

  14. Chicken line-dependent mortality after experimental infection with three type IIxIII recombinant Toxoplasma gondii clones.

    Science.gov (United States)

    Schares, G; Herrmann, D C; Maksimov, P; Matzkeit, B; Conraths, F J; Moré, G; Preisinger, R; Weigend, S

    2017-09-01

    Three genetically different clones of Toxoplasma gondii, also different in mouse virulence, were studied by experimental infection in chickens. For the experiments, four chicken lines were used, which differed in phylogenetic origin and performance level: two white egg layer lines, one with high laying performance (WLA), one with low (R11) and two brown layer lines, also displaying high (BLA) and low (L68) egg number. Chickens were intraperitoneally infected with three different T. gondii isolates representing type IIxIII recombinant clones, i.e. showing both, type II- and type III-specific alleles. These clones (K119/2 2C10, B136/1 B6H6, K119/2 A7) had exhibited virulence differences in a mouse model. In chickens, a significantly higher mortality was observed in white layer lines, but not in brown layer lines, suggesting that differences in the phylogenetic background may influence the susceptibility of chickens for toxoplasmosis. In addition, antibody (IgY) levels varied in surviving chickens at 31 days post infection. While low to intermediate antibody levels were observed in white layers, intermediate to high levels were measured in brown layers. Infection with a T. gondii clone showing low chicken virulence resulted in higher antibody levels in all chicken lines compared to infection with T. gondii clones of intermediate or high chicken virulence. This was in agreement with the parasite load as determined by real-time PCR. Overall, results show that progeny resulting from natural sexual recombination of T. gondii clonal lineages, may differ in their virulence for mice and chickens. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Construction of the recombinant vector carrying herpes simplex virus thymidine kinase and cytokine genes expressed in cell line Tca8113

    Institute of Scientific and Technical Information of China (English)

    JI Guang-hui; ZOU Jing-zhi; QU Le; YUE Ying; KUAI Jian-ke

    2004-01-01

    Objective: To construct expression vector containing fusion genes of herpes simplex virus thymidine kinase(Hsv-tk), Interleukin-2(IL-2) with internal ribosome entry sites(IRES), and to assess their expression in cell lineTca8113. Methods: IL-2 cDNA was obtained by reverse transcription. Hsv-tk, IL-2 and IRES genes were amplified by PCR. The purified amplification products were inserted into pGEM-T-Easy, and transformed into E. coli JM109. The purified recombinant plasmids were identified by restriction endonucleases. The recombinant plasmids were digested and pEGFPN3 were linearized, DNA fragments of Hsv-tk, IRES and IL-2 were ligated into linearized pEGFP-N3, and then transferred into E. coli JM109. The recombinant tk-IL-2 genes were cloned separately and introduced into the expression vector pEGFPN3 containing GFP. The recombinant vectors were identified by their restriction sites through PCR. The plasmids pEGFP-TI was also transfected into Tca8113 cells by calcium phosphate method for the expression of fusion proteins. Fusion genes expressing vector PL(TI)SN was generated by the fusion of HSV-tk, IRES and IL-2 with the use of DNA recombination technology. The recombinant retroviruses were transferred into Tca8113 cells by lipofectamine. The positive clones were obtained after G418 selection and named Tca/TI respectively. Results: The pEGFP-TI pasmid was identified respectively by restriction endonucleases, and their fragment sizes were 1 120 bp and 450 bp. The pEGFP-TI pasmid as templates were amplified respectively by PCR, and their PCR products were 1 120 bp and 450 bp. The pEGFP-TI vectors were used to transfect Tca8113 cell, and the cells with fluorescence accounted for 60 % of the total amount. Conclusion: pFGFP- tk- IRES- IL-2 expressing vector is easy to assess the expression of tk-IRES-IL-2-GFP fusion protein localization in transfected cells. The successful construction of expressing vector containing fusion genes of Hsv-tk, IRES and IL-2 may be

  16. Water Masers in the Andromeda Galaxy: I. A Survey for Water Masers, Ammonia, and Hydrogen Recombination Lines

    CERN Document Server

    Darling, Jeremy; Amiri, Nikta; Lawrence, Kelsey

    2016-01-01

    We report the results of a Green Bank Telescope survey for water masers, ammonia (1,1) and (2,2), and the H66-alpha recombination line toward 506 luminous compact 24 micron-emitting regions in the Andromeda Galaxy (M31). We include the 206 sources observed in the Darling (2011) water maser survey for completeness. The survey was sensitive enough to detect any maser useful for ~10 microarcsecond/yr astrometry. No new water masers, ammonia lines, or H66-alpha recombination lines were detected individually or in spectral stacks reaching rms noise levels of ~3 mJy and ~0.2 mJy, respectively, in 3.1-3.3 km/s channels. The lack of detections in individual spectra and in the spectral stacks is consistent with Galactic extrapolations. Contrary to previous assertions, there do not seem to be additional bright water masers to be found in M31. The strong variability of water masers may enable new maser detections in the future, but variability may also limit the astrometric utility of known (or future) masers since flar...

  17. Combination of FACS and homologous recombination for the generation of stable and high-expression engineered cell lines.

    Directory of Open Access Journals (Sweden)

    Lei Shi

    Full Text Available Traditionally, cell line generation requires several months and involves screening of over several hundred cell clones for high productivity before dozens are selected as candidate cell lines. Here, we have designed a new strategy for the generation of stable and high-expression cell lines by combining homologous recombination (HR and fluorescence-activated cell sorting (FACS. High expression was indicated by the expression of secreted green fluorescent protein (SEGFP. Parental cell lines with the highest expression of SEGFP were then selected by FACS and identified by stability analysis. Consequently, HR vectors were constructed using the cassette for SEGFP as the HR region. After transfecting the HR vector, the cells with negative SEGFP expression were enriched by FACS. The complete exchange between SEGFP and target gene (TNFR-Fc cassettes was demonstrated by DNA analysis. Compared with the traditional method, by integrating the cassette containing the gene of interest into the pre-selected site, the highest producing cells secreted a more than 8-fold higher titer of target protein. Hence, this new strategy can be applied to isolated stable cell lines with desirable expression of any gene of interest. The stable cell lines can rapidly produce proteins for researching protein structure and function and are even applicable in drug discovery.

  18. Modelling of spectral lines emitted by hydrogen isotopes for ionising and recombining plasma conditions of Tokamak edges

    Energy Technology Data Exchange (ETDEWEB)

    Koubiti, M.; Marandet, Y.; Godbert-Mouret, L.; Stamm, R.; Touati, K. [Physique des Interactions Ioniques et Moleculaires, UMR 6633 CNRS/Universite de Provence, centre de Saint-Jerome, Marseille (Spain); Capes, H.; Escarguel, A.; Guirlet, R.; Michelis, C. De [Departement de Recherches sur la Fusion Controlee, Association EURATOM-CEA Cadrache, Saint Paul lez Durance (France)

    2001-07-01

    The plasma in the periphery of a magnetic fusion device plays an important role in the spread of particle and heat power. To optimise its role, it is necessary to characterize the edge plasma and understand all the interaction processes between the plasma constituents (ions, electrons and neutrals), and also their interactions with the device walls. For that purpose, high-resolution passive emission spectroscopy is well suitable since it is a non-intrusive method However, edge plasma diagnostics based on this method requires the accurate modelling of the observed spectra. A lineshape model initially developed for Stark broadening has been recently updated to include the Zeeman effect. The synthetic line profiles can be convolved with a Gaussian or a Lorentzian to account for the Doppler broadening and the instrumental function. For ionising conditions of edge plasmas such as those realized in front of the neutraliser plates (NP) of the Tore-Supra (TS) Ergodic Divertor where the electron temperature and density are usually higher than 10 eV and lower than 10{sup 19} m{sup -3}, the electron excitation of deuterium atoms dominates the recombination and only the first lines of the Balmer series are measured By fitting and analysing the spectrum of the deuterium Balmer {alpha} line (D{alpha}) emitted at different radial positions, information on the edge neutral populations can be obtained. In particular the neutral pro' auction mechanism and the dominant neutral relaxation processes can be identified from Doppler profile analysis. For this purpose we have analysed the lineshape of (D{alpha}) emitted in front of an equatorial NP of the ergodic diverter by a plasma region extending up to 2 cm radially away from the NP surface. For recombining plasma conditions such as Hose obtained in axisymmetric divertors, the electron temperature and density are respectively lower than 1 eV and higher than 10{sup 20} m{sup -3}, the volume recombination dominates the excitation

  19. Construction of Recombinant Bacmid Containing M2e-Ctxb and Producing the Fusion Protein in Insect Cell Lines

    Science.gov (United States)

    Mirzaei, Nima; Mokhtari Azad, Talat; Nategh, Rakhshandeh; Soleimanjahi, Hoorieh; Amirmozafari, Nour

    2014-01-01

    Background: Sequence variations in glycoproteins of influenza virus surface impel us to design new candidate vaccines yearly. Ectodomain of influenza M2 protein is a surface and highly conserved protein. M2e in influenza vaccines may eliminate the need for changing vaccine formulation every year. Objectives: In this study, a recombinant baculovirus containing M2e and cholera toxin subunit B fusion gene was generated with transposition process to express in large amounts in insect cell lines. Materials and Methods: M2e-ctxB fusion gene was created and cloned into pFastBac HT. The recombinant vector was transformed into DH10Bac cells to introduce the fusion gene into the bacmid DNA via a site-specific transposition process. The recombinant bacmid was then extracted from white colonies and further analyzed using PCR, DNA sequence analyzing, and indirect immunofluorescence assay. Results: PCR and DNA sequence analyzing results showed that the fusion gene was constructed as a single open reading frame and was successfully inserted into bacmid DNA. Moreover, indirect immunofluorescence results showed that the fusion gene was successfully expressed. Conclusions: Baculovirus expression vector system is valuable to produce M2e based influenza vaccines due to its simple utilization and ease of target gene manipulation. The expressed protein in such systems can improve the evaluating process of new vaccination strategies. PMID:24719728

  20. Purification of Recombinant Ebola Virus Glycoprotein and VP40 from a Human Cell Line

    Science.gov (United States)

    2017-01-01

    be the only viral protein responsible for entry into host cells (Takada et al., 1997; Wool- Lewis and Bates, 1998), and thus, it is a logical target...maltose binding protein to assist in protein folding for purification. This is connected to VP40 at the C-terminus by a 14 amino acid linker, which...PAGE image of resultant fractions. The expected molecular weight of recombinant EBV GP, based solely on amino acid composition, was 73 kDa. Because

  1. On-line monitoring of respiration in recombinant-baculovirus infected and uninfected insect cell bioreactor cultures.

    Science.gov (United States)

    Kamen, A A; Bédard, C; Tom, R; Perret, S; Jardin, B

    1996-04-05

    Respiration rates in Spodoptera frugiperda (Sf-9) cell bioreactor cultures were successfully measured on-line using two methods: The O(2) uptake rate (OUR) was determined using gas phase pO(2) values imposed by a dissolved oxygen controller and the CO(2) evolution rate (CER) was measured using an infrared detector. The measurement methods were accurate, reliable, and relatively inexpensive. The CER was routinely determined in bioreactor cultures used for the production of several recombinant proteins. Simple linear relationships between viable cell densities and both OUR and CER in exponentially growing cultures were used to predict viable cell density. Respiration measurements were also used to follow the progress of baculoviral infections in Sf-9 cultures. Infection led to increases in volumetric and per-cell respiration rates. The relationships between respiration and several other culture parameters, including viable cell density, cell protein, cell volume, glucose consumption, lactate production, viral titer, and recombinant beta-galactosidase accumulation, were examined. The extent of the increase in CER following infection and the time postinfection at which maximum CER was attained were negatively correlated with the multiplicity of infection (MOI) at multiplicities below the level required to infect all the cells in a culture. Delays in the respiration peak related to the MOI employed were correlated with delays in the peak in recombinant protein accumulation. DO levels in the range 5-100% did not exert any major effects on viable cell densities, CER, or product titer in cultures infected with a baculovirus expressing recombinant beta-galactosidase.

  2. On the Ionization of Luminous WMAP Sources in the Galaxy : Constraints from He Recombination Line Observations with the GBT

    CERN Document Server

    Roshi, D Anish; Rosero, Viviana; Vaddi, Sravani

    2012-01-01

    The Wilkinson Microwave Anisotropy Probe (WMAP) free-free foreground emission map is used to identify diffuse ionized regions (DIR) in the Galaxy (Rahman & Murray 2010). It has been found that the 18 most luminous WMAP sources produce more than half of the total ionizing luminosity of the Galaxy. We observed radio recombination lines (RRLs) toward the luminous WMAP source G49.75-0.45 with the Green Bank Telescope near 1.4 GHz. Hydrogen RRL is detected toward the source but no helium line is detected, implying that n_He+/n_H+ ~10 times to explain the observations. If selective absorption of He- ionizing photons by dust is causing this additional attenuation, then the ratio of dust absorption cross sections for He- and H- ionizing photons should be > ~6.

  3. A survey of Radio Recombination Lines using Ooty Radio Telescope at 328 MHz in the Inner Galaxy

    CERN Document Server

    Baddi, Raju

    2012-01-01

    A survey of radio recombination lines in the Galactic plane with longitude $-32^o < l < +80^o$ and latitude $b<\\pm3^o$ using Ooty Radio Telescope(ORT) at 328 MHz has been reported. ORT observations were made using a New Digital Backend(NDB) augmented to it recently. With NDB ORT had a beam of $2^o.3 \\times 2^o.2 sec(\\delta)$ and a passband of $\\sim$1 MHz in the spectral line mode. The above mentioned Galactic region was divided into $\\sim 2^o \\times 2^o$ patches with the ORT beam pointed to the center. The ORT observations form a study of distribution of extended low-density warm-ionized medium(ELDWIM) in the inner Galaxy using H271$\\alpha$ RL. By obtaining kinematical distances using $V_{LSR}$ of the H271$\\alpha$ RLs the distribution of ELDWIM clouds within the inner Galaxy has been deduced for the region given above.

  4. Radio continuum and radio recombination line observations of Sagittarius B1 and G0.6-0.0

    Science.gov (United States)

    Mehringer, David M.; Yusef-Zadeh, F.; Palmer, Patrick; Goss, W. M.

    1992-01-01

    Continuum emission and H110-alpha recombination line emission from Sgr B1 and G0.6-0.0 have been observed using the VLA. It is shown that Sgr B1 is a region of great complexity, both spatially and kinematically. The continuum observations show that this region is dominated by many extended features rather than compact sources. On the other hand, Sgr B2 is dominated by several ultracompact H II regions. The two regions may be in different stages of evolution, with Sgr B1 being older, perhaps by as much as 0 exp 6 yrs. The recombination line study shows that Sgr B1 is composed of two distinct kinematic regions, a simple western one and a more complex eastern one. G0.6-0.0 is a region composed of at least four ultracompact H II regions that is situated between Sgr B1 and Sgr B2. There is an arc of ionized gas that lies to the east and to the south of these compact regions. The velocity of G0.6-0.0 is intermediate between those of Sgr B1 and Sgr B2. These facts strengthen the argument that these regions are physically associated.

  5. Effect of salinity on zinc uptake by Brassica juncea.

    Science.gov (United States)

    Novo, Luís A B; Covelo, Emma F; González, Luís

    2014-01-01

    Salinity is a major worldwide problem that affects agricultural soils and limits the reclamation of contaminated sites. Despite the large number of research papers published about salt tolerance in Brassica juncea L., there are very few accounts concerning the influence of salinity on the uptake of trace metals. In this study, B. juncea plants divided through soil sets comprising 0, 900 and 1800 mg Zn kg(-1), were treated with solutions containing 0, 60 and 120 mmol L(-1) of NaCl, with the purpose of observing the effect of salt on Zn uptake, and some physiological responses throughout the 90 days experiment. Increasing concentrations of NaCl and Zn produced a decline in the ecophysiological and biochemical properties of the plants, with observable synergistic effects on parameters like shoot dry weight, leaf area, or photochemical efficiency. Nevertheless, plants treated with 60 mmol L(-1) of NaCl accumulated striking harvestable amounts of Zn per plant that largely exceed those reported for Thlaspi caerulescens. It was concluded that salinity could play an important role on the uptake of Zn by B. juncea. The potential mechanisms behind these results are discussed, as well as the implications for phytoremediation of Zn on saline and non-saline soils.

  6. Inheritance of rapeseed (Brassica napus)-specific RAPD markers and a transgene in the cross B.juncea x (B.juncea x B.napus)

    DEFF Research Database (Denmark)

    Frello, S.; Hansen, K.R.; Jensen, J.

    1995-01-01

    , with B. juncea as the female parent, was successful both in controlled crosses and spontaneously in the field. The controlled backcrossing of selected hybrids to B. juncea, again with B. juncea as the female parent, also resulted in many seeds. The BC1 plants contained from 0 to 20 of the rapeseed RAPD...... markers, and the frequency of inheritance of individual RAPD markers ranged from 19% to 93%. The transgene was found in 52% of the plants analyzed. Five synteny groups of RAPD markers were identified. In the hybrids pollen fertility was 0-28%. The hybrids with the highest pollen fertility were selected...

  7. Eliminating tyrosine sequence variants in CHO cell lines producing recombinant monoclonal antibodies.

    Science.gov (United States)

    Feeney, Lauren; Carvalhal, Veronica; Yu, X Christopher; Chan, Betty; Michels, David A; Wang, Yajun Jennifer; Shen, Amy; Ressl, Jan; Dusel, Brendon; Laird, Michael W

    2013-04-01

    Amino acid sequence variants are defined as unintended amino acid sequence changes that contribute to product variation with potential impact to product safety, immunogenicity, and efficacy. Therefore, it is important to understand the propensity for sequence variant (SV) formation during the production of recombinant proteins for therapeutic use. During the development of clinical therapeutic products, several monoclonal antibodies (mAbs) produced from Chinese Hamster Ovary (CHO) cells exhibited SVs at low levels (≤3%) in multiple locations throughout the mAbs. In these examples, the cell culture process depleted tyrosine, and the tyrosine residues in the recombinant mAbs were replaced with phenylalanine or histidine. In this work, it is demonstrated that tyrosine supplementation eliminated the tyrosine SVs, while early tyrosine starvation significantly increased the SV level in all mAbs tested. Additionally, it was determined that phenylalanine is the amino acid preferentially misincorporated in the absence of tyrosine over histidine, with no other amino acid misincorporated in the absence of tyrosine, phenylalanine, and histidine. The data support that the tyrosine SVs are due to mistranslation and not DNA mutation, most likely due to tRNA(Tyr) mischarging due to the structural similarities between tyrosine and phenylalanine.

  8. Effect of dielectronic recombination on the charge-state distribution and soft X-ray line intensity of laser-produced carbon plasma

    Indian Academy of Sciences (India)

    A Chowdhury; G P Gupta; P A Naik; P D Gupta

    2005-01-01

    The effect of dielectronic recombination in determining charge-state distribution and radiative emission from a laser-produced carbon plasma has been investigated in the collisional radiative ionization equilibrium. It is observed that the relative abundances of different ions in the plasma, and soft X-ray emission intensity get significantly altered when dielectronic recombination is included. Theoretical estimates of the relative population of CVI to CV ions and ratio of line intensity emitted from them for two representative formulations of dielectronic recombination are presented.

  9. Yield reduction in Brassica napus, B. rapa, B. juncea, and Sinapis alba caused by flea beetle (Phyllotreta cruciferae (Goeze) (Coleoptera: Chrysomelidae)) infestation in northern Idaho.

    Science.gov (United States)

    Brown, Jack; McCaffrey, Joseph P; Brown, Donna A; Harmon, Bradley L; Davis, James B

    2004-10-01

    Phyllotreta cruciferae is an important insect pest of spring-planted Brassica crops, especially during the seedling stage. To determine the effect of early season P. cruciferae infestation on seed yield, 10 genotypes from each of two canola species (Brassica napus L. and Brassica rapa L.) and two mustard species (Brassica juncea L. and Sinapis alba L.) were grown in 2 yr under three different P. cruciferae treatments: (1) no insecticide control; (2) foliar applications of endosulfan; and (3) carbofuran with seed at planting plus foliar application of carbaryl. Averaged over 10 genotypes, B. rapa showed most visible P. cruciferae injury and showed greatest yield reduction without insecticide application. Mustard species (S. alba and B. juncea) showed least visible injury and higher yield without insecticide compared with canola species (B. napus and B. rapa). Indeed, average seed yield of S. alba without insecticide was higher than either B. napus or B. rapa with most effective P. cruciferae control. Significant variation occurred within each species. A number of lines from B. napus, B. juncea, anid S. alba showed less feeding injury and yield reduction as a result of P. cruciferae infestation compared with other lines from the same species examined, thus having potential genetic background for developing resistant cultivars.

  10. QTL mapping of fruit rot resistance to the plant pathogen Phytophthora capsici in a recombinant inbred line Capsicum annuum population.

    Science.gov (United States)

    Naegele, R P; Ashrafi, H; Hill, T A; Chin-Wo, S Reyes; Van Deynze, A E; Hausbeck, M K

    2014-05-01

    Phytophthora capsici is an important pepper (Capsicum annuum) pathogen causing fruit and root rot, and foliar blight in field and greenhouse production. Previously, an F6 recombinant inbred line population was evaluated for fruit rot susceptibility. Continuous variation among lines and partial and isolate-specific resistance were found. In this study, Phytophthora fruit rot resistance was mapped in the same F6 population between Criollo del Morelos 334 (CM334), a landrace from Mexico, and 'Early Jalapeno' using a high-density genetic map. Isolate-specific resistance was mapped independently in 63 of the lines evaluated and the two parents. Heritability of the resistance for each isolate at 3 and 5 days postinoculation (dpi) was high (h(2) = 0.63 to 0.68 and 0.74 to 0.83, respectively). Significant additive and epistatic quantitative trait loci (QTL) were identified for resistance to isolates OP97 and 13709 (3 and 5 dpi) and 12889 (3 dpi only). Mapping of fruit traits showed potential linkage with few disease resistance QTL. The partial fruit rot resistance from CM334 suggests that this may not be an ideal source for fruit rot resistance in pepper.

  11. Carbon and oxygen abundances from recombination lines in low-metallicity star-forming galaxies. Implications for chemical evolution

    CERN Document Server

    Esteban, C; Carigi, L; Peimbert, M; Bresolin, F; López-Sánchez, A R; Mesa-Delgado, A

    2014-01-01

    We present deep echelle spectrophotometry of the brightest emission-line knots of the star-forming galaxies He 2-10, Mkn 1271, NGC 3125, NGC 5408, POX 4, SDSS J1253-0312, Tol 1457-262, Tol 1924-416 and the HII region Hubble V in the Local Group dwarf irregular galaxy NGC 6822. The data have been taken with the Very Large Telescope Ultraviolet-Visual Echelle Spectrograph in the 3100-10420 $\\AA$ range. We determine electron densities and temperatures of the ionized gas from several emission-line intensity ratios for all the objects. We derive the ionic abundances of C$^{2+}$ and/or O$^{2+}$ from faint pure recombination lines (RLs) in several of the objects, permitting to derive their C/H and C/O ratios. We have explored the chemical evolution at low metallicities analysing the C/O vs. O/H, C/O vs. N/O and C/N vs. O/H relations for Galactic and extragalactic HII regions and comparing with results for halo stars and DLAs. We find that HII regions in star-forming dwarf galaxies occupy a different locus in the C/O...

  12. The Lick AGN Monitoring Project: Reverberation Mapping of Optical Hydrogen and Helium Recombination Lines

    CERN Document Server

    Bentz, Misty C; Barth, Aaron J; Yoshii, Yuzuru; Woo, Jong-Hak; Wang, Xiaofeng; Treu, Tommaso; Thornton, Carol E; Street, Rachel A; Steele, Thea N; Silverman, Jeffrey M; Serduke, Frank J D; Sakata, Yu; Minezaki, Takeo; Malkan, Matthew A; Li, Weidong; Lee, Nicholas; Hiner, Kyle D; Hidas, Marton G; Greene, Jenny E; Gates, Elinor L; Ganeshalingam, Mohan; Filippenko, Alexei V; Canalizo, Gabriela; Bennert, Vardha Nicola; Baliber, Nairn

    2010-01-01

    We have recently completed a 64-night spectroscopic monitoring campaign at the Lick Observatory 3-m Shane telescope with the aim of measuring the masses of the black holes in 12 nearby (z < 0.05) Seyfert 1 galaxies with expected masses in the range ~10^6-10^7M_sun and also the well-studied nearby active galactic nucleus (AGN) NGC 5548. Nine of the objects in the sample (including NGC 5548) showed optical variability of sufficient strength during the monitoring campaign to allow for a time lag to be measured between the continuum fluctuations and the response to these fluctuations in the broad Hbeta emission, which we have previously reported. We present here the light curves for the Halpha, Hgamma, HeII 4686, and HeI 5876 emission lines and the time lags for the emission-line responses relative to changes in the continuum flux. Combining each emission-line time lag with the measured width of the line in the variable part of the spectrum, we determine a virial mass of the central supermassive black hole fro...

  13. Stability of transgene expression, field performance and recombination breeding of transformed barley lines

    DEFF Research Database (Denmark)

    Horvath, H.; Jensen, L.G.; Wong, O.T.

    2001-01-01

    in homozygous transgenic T-3 plants, and these remained constant over a 3-year period. In micro-malting experiments, the heat-stable enzyme reached levels of up to 1.4 mug.mg(-1) protein and survived kiln drying at levels of 70-100%. In the field trials of 1997 and 1998 the transgenic lines had a reduced 1000...... lines yielded approximately 6 t.ha(-1) and Golden Promise 7.7 t.ha(-1). Cross-breeding was carried out to transfer the transgene into a more suitable genetic background. Crosses of the semi-dwarf ari-e mutant Golden Promise gave rise to the four morphological phenotypes nutans, high erect, erect...... transformants were observed in some F-4 lines homozygous for the morphological phenotypes and for the transgene. In the case of a homozygous nutans line, the transgenic plants had a higher 1000-grain weight than those lacking the transgene. Like mutants providing useful output traits, transgenic plants...

  14. Effects of glutamine and asparagine on recombinant antibody production using CHO-GS cell lines.

    Science.gov (United States)

    Xu, Ping; Dai, Xiao-Ping; Graf, Erica; Martel, Richard; Russell, Reb

    2014-01-01

    A unique and nontraditional approach using glutamine and asparagine supplements for CHO-glutamine synthetase (GS) cell lines was studied. In our experiments, we found that a decrease in pH and an increase in cell death occurred in production phase of a GS cell line, leading to reduced antibody expression and lower antibody yields. The experimental results and the statistical analysis (ANOVA) indicated that additions of glutamine and asparagine in the basal and feed media were effective to buffer the cell culture pH, reduce lactate generation, maintain a higher cell viability profile, and improve antibody productivity. In bench-top bioreactors, glutamine and asparagine supplementation helped to prevent cell death, improve antibody yield, and reduce base usage. Glutamine is normally excluded from culture media for GS cell lines to prevent the bypass of selection pressure. In this study, however, the addition of glutamine did not affect cell population homogeneity, protein quality, or decrease antibody yield of two GS cell lines.

  15. Transgenic tobacco overexpressing Brassica juncea HMG-CoA synthase 1 shows increased plant growth, pod size and seed yield.

    Science.gov (United States)

    Liao, Pan; Wang, Hui; Wang, Mingfu; Hsiao, An-Shan; Bach, Thomas J; Chye, Mee-Len

    2014-01-01

    Seeds are very important not only in the life cycle of the plant but they represent food sources for man and animals. We report herein a mutant of 3-hydroxy-3-methylglutaryl-coenzyme A synthase (HMGS), the second enzyme in the mevalonate (MVA) pathway that can improve seed yield when overexpressed in a phylogenetically distant species. In Brassica juncea, the characterisation of four isogenes encoding HMGS has been previously reported. Enzyme kinetics on recombinant wild-type (wt) and mutant BjHMGS1 had revealed that S359A displayed a 10-fold higher enzyme activity. The overexpression of wt and mutant (S359A) BjHMGS1 in Arabidopsis had up-regulated several genes in sterol biosynthesis, increasing sterol content. To quickly assess the effects of BjHMGS1 overexpression in a phylogenetically more distant species beyond the Brassicaceae, wt and mutant (S359A) BjHMGS1 were expressed in tobacco (Nicotiana tabacum L. cv. Xanthi) of the family Solanaceae. New observations on tobacco OEs not previously reported for Arabidopsis OEs included: (i) phenotypic changes in enhanced plant growth, pod size and seed yield (more significant in OE-S359A than OE-wtBjHMGS1) in comparison to vector-transformed tobacco, (ii) higher NtSQS expression and sterol content in OE-S359A than OE-wtBjHMGS1 corresponding to greater increase in growth and seed yield, and (iii) induction of NtIPPI2 and NtGGPPS2 and downregulation of NtIPPI1, NtGGPPS1, NtGGPPS3 and NtGGPPS4. Resembling Arabidopsis HMGS-OEs, tobacco HMGS-OEs displayed an enhanced expression of NtHMGR1, NtSMT1-2, NtSMT2-1, NtSMT2-2 and NtCYP85A1. Overall, increased growth, pod size and seed yield in tobacco HMGS-OEs were attributed to the up-regulation of native NtHMGR1, NtIPPI2, NtSQS, NtSMT1-2, NtSMT2-1, NtSMT2-2 and NtCYP85A1. Hence, S359A has potential in agriculture not only in improving phytosterol content but also seed yield, which may be desirable in food crops. This work further demonstrates HMGS function in plant reproduction

  16. A Cell Line Producing Recombinant Nerve Growth Factor Evokes Growth Responses in Intrinsic and Grafted Central Cholinergic Neurons

    Science.gov (United States)

    Ernfors, Patrik; Ebendal, Ted; Olson, Lars; Mouton, Peter; Stromberg, Ingrid; Persson, Hakan

    1989-06-01

    The rat β nerve growth factor (NGF) gene was inserted into a mammalian expression vector and cotransfected with a plasmid conferring resistance to neomycin into mouse 3T3 fibroblasts. From this transfection a stable cell line was selected that contains several hundred copies of the rat NGF gene and produces excess levels of recombinant NGF. Such genetically modified cells were implanted into the rat brain as a probe for in vivo effects of NGF on central nervous system neurons. In a model of the cortical cholinergic deficits in Alzheimer disease, we demonstrate a marked increase in the survival of, and fiber outgrowth from, grafts of fetal basal forebrain cholinergic neurons, as well as stimulation of fiber formation by intact adult intrinsic cholinergic circuits in the cerebral cortex. Adult cholinergic interneurons in intact striatum also sprout vigorously toward implanted fibroblasts. Our results suggest that this model has implications for future treatment of neurodegenerative diseases.

  17. Inheritance of Late Maturity α-amylase (LMA) in a Recombinant Inbreed Lines of Chuanmai 42 Crossed with Chuannong 16

    Institute of Scientific and Technical Information of China (English)

    YANG En-nian; ZHANG Yu; LI Jun; TANG Yong-lu; YANG Wu-yun; ZOU Yu-chun

    2011-01-01

    [ Objective] To study the inheritance and possible molecular marker of α-LMA and their relations to falling number. [ Method] Recombined inbreed lines (RILS) of Chuanmai 42 (LMA genotype) crossed with Chuannong 16 (no LMA) were employed to test the late maturity α-amylase (LMA) by EL ISA kits, the Falling Number (FN), and three QTL, SSR markers ( Xgwm 577, Xwmc2.76, Xwmc273) of LMA gene.[ Result] Results suggested that LMA was controlled by a single, and the three SSR markers seemed to be not efficient to test LMA gene in this RILS. LMA was an important factor on FN, meanwhile FN was also affected by other factors. [Conclusion] The study could provide references for the breed of quality wheat.

  18. Heterosis as investigated in terms of polyploidy and genetic diversity using designed Brassica juncea amphiploid and its progenitor diploid species.

    Directory of Open Access Journals (Sweden)

    Payal Bansal

    Full Text Available Fixed heterosis resulting from favorable interactions between the genes on their homoeologous genomes in an allopolyploid is considered analogous to classical heterosis accruing from interactions between homologous chromosomes in heterozygous plants of a diploid species. It has been hypothesized that fixed heterosis may be one of the causes of low classical heterosis in allopolyploids. We used Indian mustard (Brassica juncea, 2n = 36; AABB as a model system to analyze this hypothesis due to ease of its resynthesis from its diploid progenitors, B. rapa (2n = 20; AA and B. nigra (2n = 16; BB. Both forms of heterosis were investigated in terms of ploidy level, gene action and genetic diversity. To facilitate this, eleven B. juncea genotypes were resynthesized by hybridizing ten near inbred lines of B. rapa and nine of B. nigra. Three half diallel combinations involving resynthesized B. juncea (11×11 and the corresponding progenitor genotypes of B. rapa (10×10 and B. nigra (9×9 were evaluated. Genetic diversity was estimated based on DNA polymorphism generated by SSR primers. Heterosis and genetic diversity in parental diploid species appeared not to predict heterosis and genetic diversity at alloploid level. There was also no association between combining ability, genetic diversity and heterosis across ploidy. Though a large proportion (0.47 of combinations showed positive values, the average fixed heterosis was low for seed yield but high for biomass yield. The genetic diversity was a significant contributor to fixed heterosis for biomass yield, due possibly to adaptive advantage it may confer on de novo alloploids during evolution. Good general/specific combiners at diploid level did not necessarily produce good general/specific combiners at amphiploid level. It was also concluded that polyploidy impacts classical heterosis indirectly due to the negative association between fixed heterosis and classical heterosis.

  19. Biological insights into the expression of translation initiation factors from recombinant CHOK1SV cell lines and their relationship to enhanced productivity.

    Science.gov (United States)

    Mead, Emma J; Masterton, Rosalyn J; Feary, Marc; Obrezanova, Olga; Zhang, Lin; Young, Robert; Smales, C Mark

    2015-12-15

    Translation initiation is on the critical pathway for the production of monoclonal antibodies (mAbs) by mammalian cells. Formation of a closed loop structure comprised of mRNA, a number of eukaryotic initiation factors (eIFs) and ribosomal proteins has been proposed to aid re-initiation of translation and therefore increase global translational efficiency. We have determined mRNA and protein levels of the key components of the closed loop, eIFs (eIF3a, eIF3b, eIF3c, eIF3h, eIF3i and eIF4G1), poly(A)-binding protein (PABP) 1 and PABP-interacting protein 1 (PAIP1), across a panel of 30 recombinant mAb-producing GS-CHOK1SV cell lines with a broad range of growth characteristics and production levels of a model recombinant mAb. We have used a multi-level statistical approach to investigate the relationship between key performance indicators (cell growth and recombinant antibody productivity) and the intracellular amounts of target translation initiation factor proteins and the mRNAs encoding them. We show that high-producing cell lines maintain amounts of the translation initiation factors involved in the formation of the closed loop mRNA, maintaining these proteins at appropriate levels to deliver enhanced recombinant protein production. We then utilize knowledge of the amounts of these factors to build predictive models for and use cluster analysis to identify, high-producing cell lines. The present study therefore defines the translation initiation factor amounts that are associated with highly productive recombinant GS-CHOK1SV cell lines that may be targets for screening highly productive cell lines or to engineer new host cell lines with the potential for enhanced recombinant antibody productivity.

  20. Transgenic Brassica juncea plants expressing MsrA1, a synthetic cationic antimicrobial peptide, exhibit resistance to fungal phytopathogens.

    Science.gov (United States)

    Rustagi, Anjana; Kumar, Deepak; Shekhar, Shashi; Yusuf, Mohd Aslam; Misra, Santosh; Sarin, Neera Bhalla

    2014-06-01

    Cationic antimicrobial peptides (CAPs) have shown potential against broad spectrum of phytopathogens. Synthetic versions with desirable properties have been modeled on these natural peptides. MsrA1 is a synthetic chimera of cecropin A and melittin CAPs with antimicrobial properties. We generated transgenic Brassica juncea plants expressing the msrA1 gene aimed at conferring fungal resistance. Five independent transgenic lines were evaluated for resistance to Alternaria brassicae and Sclerotinia sclerotiorum, two of the most devastating pathogens of B. juncea crops. In vitro assays showed inhibition by MsrA1 of Alternaria hyphae growth by 44-62 %. As assessed by the number and size of lesions and time taken for complete leaf necrosis, the Alternaria infection was delayed and restricted in the transgenic plants with the protection varying from 69 to 85 % in different transgenic lines. In case of S. sclerotiorum infection, the lesions were more severe and spread profusely in untransformed control compared with transgenic plants. The sclerotia formed in the stem of untransformed control plants were significantly more in number and larger in size than those present in the transgenic plants where disease protection of 56-71.5 % was obtained. We discuss the potential of engineering broad spectrum biotic stress tolerance by transgenic expression of CAPs in crop plants.

  1. Mutagenesis and homologous recombination in Drosophila cell lines using CRISPR/Cas9

    Directory of Open Access Journals (Sweden)

    Andrew R. Bassett

    2013-12-01

    We have applied the CRISPR/Cas9 system to Drosophila S2 cells to generate targeted genetic mutations in more than 85% of alleles. By targeting a constitutive exon of the AGO1 gene, we demonstrate homozygous mutation in up to 82% of cells, thereby allowing the study of genetic knockouts in a Drosophila cell line for the first time. We have shown that homologous gene targeting is possible at 1–4% efficiency using this system, allowing for the construction of defined insertions and deletions. We demonstrate that a 1 kb homology arm length is optimal for integration by homologous gene targeting, and demonstrate its efficacy by tagging the endogenous AGO1 protein. This technology enables controlled genetic manipulation in Drosophila cell lines, and its simplicity offers the opportunity to study cellular phenotypes genome-wide.

  2. THOR - The HI, OH, Recombination Line Survey of the Milky Way - HI observations of the giant molecular cloud W43

    Science.gov (United States)

    Bihr, Simon; Beuther, Henrik; Johnston, Katharine; Henning, Thomas; Ott, Juergen; Brunthaler, Andreas; THOR Collaboration

    2015-08-01

    To study the atomic, molecular and ionised emission of Giant Molecular Clouds (GMCs) in the Milky Way, we have initiated a Large Program with the VLA: 'THOR - The HI, OH, Recombination Line survey of the Milky Way'. We map the 21cm HI line, 4 OH lines, 19 Hα recombination lines and the continuum from 1-2GHz of a significant fraction of the Milky Way (l=15-67°, |b|thin with properties such as the column density calculated under this assumption. While this approach gives reasonable results for regions of low-mass star-formation, it is not sufficient to describe the atomic gas in close proximity to GMCs. In my talk, I will present a method using strong continuum sources to measure the optical depth, and thus correct the HI 21cm emission for optical depth effects and weak diffuse continuum emission. Our analysis puts a lower limit of M~6.6x106 Msun on the HI mass associated with the W43 GMC, which is a factor of 2.4 larger than the mass obtained using the optically thin assumption. The HI column densities reach NHI~150 Msun pc-2 ~ 1.9x1022 cm-2, which is an order of magnitude higher than seen in low mass star formation regions. This result challenges theoretical models that predict a threshold for the HI column density of ~10 Msun pc-2, at which the formation of molecular hydrogen should set in. Furthermore, we assume an elliptical layered structure for W43 to estimate the particle density profile. The HI particle density shows a linear decrease toward the centre of W43 and the molecular hydrogen, traced via dust observations with Herschel, shows an exponential increase toward the centre. While at the cloud edge atomic and molecular hydrogen are well mixed, the centre of the cloud is dominated by H2. We do not identify a sharp transition between hydrogen in atomic and molecular form. Our results are an important characterization of the atomic to molecular hydrogen transition in an extreme environment that challenges current theoretical models.

  3. Performance of Sorghum Recombinant Inbred Lines (RIL developed for rain-fed areas of Sudan

    Directory of Open Access Journals (Sweden)

    Hassan El Tayieb Ibrahim

    2015-01-01

    Full Text Available Sorghum (Sorghum bicolor (L Moench is the most widely produced and consumed cereal crop in Sudan. However, productivity is low since the crop is produced in favorable and unfavorable environments where the crop suffers from drought stresses at different growth stages. In the present study, six sorghum inbred lines developed by local breeding program and two commercial checks were evaluated for grain yield potential, yield stability, some important agronomic characters and grain quality properties. Series variety trials were conducted at Elobeid and Suki, Damazin areas of Sudan, during three consecutive rainy seasons. The selected locations represent low, medium and high rainfall areas of Sudan. The trial was laid out in a randomized complete block design. The results revealed that mean squares of genotypes, seasons, locations, location x season, location x genotypes, season x genotypes and season x location x genotype interactions were highly significant (P=0.01 for grain yield. The sorghum genotypes Edo 34-23-4, Edo 26-18 and Edo 16-dwarf produced substantially higher grain yields than commercial checks and the trial mean. Their percentage yield increase ranged from 5% to 75% over commercial checks. The stability analysis revealed that the above mentioned Edo-genotypes had high yield potentials and were stable across a wide range of agricultural conditions. Moreover, the same Edo-lines showed early maturing compared to selected commercial checks and also the Edo-lines possessed good food grains and were market preferred and acceptable for making quality kisra (fermented sorghum pancake-like flatbread. The Edo developed lines also possess the acceptable grain quality in addition to moderate physical grain characteristics such as protein content, fat acidity and moisture content.

  4. Chemical inhibition of autophagy: Examining its potential to increase the specific productivity of recombinant CHO cell lines.

    Science.gov (United States)

    Baek, Eric; Kim, Che Lin; Kim, Mi Gyeom; Lee, Jae Seong; Lee, Gyun Min

    2016-09-01

    Chinese hamster ovary (CHO) cells activate and undergo apoptosis and autophagy for various environmental stresses. Unlike apoptosis, studies on increasing the production of therapeutic proteins in CHO cells by targeting the autophagy pathway are limited. In order to identify the effects of chemical autophagy inhibitors on the specific productivity (qp ), nine chemical inhibitors that had been reported to target three different phases of autophagy (metformin, dorsomorphin, resveratrol, and SP600125 against initiation and nucleation; 3-MA, wortmannin, and LY294002 against elongation, and chloroquine and bafilomycin A1 against autophagosome fusion) were used to treat three recombinant CHO (rCHO) cell lines: the Fc-fusion protein-producing DG44 (DG44-Fc) and DUKX-B11 (DUKX-Fc) and antibody-producing DG44 (DG44-Ab) cell lines. Among the nine chemical inhibitors tested, 3-MA, dorsomorphin, and SP600125 significantly increased the qp of DG44-Fc and DUKX-Fc. In contrast, for DG44-Ab, only 3-MA significantly increased the qp . The autophagy-inhibiting activity of the nine chemical inhibitors on the rCHO cell lines was evaluated through Western blot analysis and flow cytometry. Unexpectedly, some chemical inhibitors did not exhibit any apparent inhibition activity on autophagy. The chemical inhibitors that enhanced the qp , 3-MA, dorsomorphin, and SP600125, exhibited instead an increased autophagic flux. Taken all together, the chemical inhibition of autophagy was not effective in increasing the qp in rCHO cell lines and the positive effect of 3-MA, dorsomorphin, and SP600125 on the qp was not due to the inhibition of autophagy. Biotechnol. Bioeng. 2016;113: 1953-1961. © 2016 Wiley Periodicals, Inc.

  5. Plant growth regulators enhance gold uptake in Brassica juncea.

    Science.gov (United States)

    Kulkarni, Manoj G; Stirk, Wendy A; Southway, Colin; Papenfus, Heino B; Swart, Pierre A; Lux, Alexander; Vaculík, Marek; Martinka, Michal; Van Staden, Johannes

    2013-01-01

    The use of plant growth regulators is well established and they are used in many fields of plant science for enhancing growth. Brassica juncea plants were treated with 2.5, 5.0 and 7.5 microM auxin indole-3-butyric acid (IBA), which promotes rooting. The IBA-treated plants were also sprayed with 100 microM gibberellic acid (GA3) and kinetin (Kin) to increase leaf-foliage. Gold (I) chloride (AuCl) was added to the growth medium of plants to achieve required gold concentration. The solubilizing agent ammonium thiocyanate (1 g kg(-1)) (commonly used in mining industries to solubilize gold) was added to the nutrient solution after six weeks of growth and, two weeks later, plants were harvested. Plant growth regulators improved shoot and root dry biomass of B. juncea plants. Inductively Coupled Plasma Optical Emission Spectrometry analysis showed the highest Au uptake for plants treated with 5.0 microM IBA. The average recovery of Au with this treatment was significantly greater than the control treatment by 45.8 mg kg(-1) (155.7%). The other IBA concentrations (2.5 and 7.5 microM) also showed a significant increase in Au uptake compared to the control plants by 14.7 mg kg(-1) (50%) and 42.5 mg kg(-1) (144.5%) respectively. A similar trend of Au accumulation was recorded in the roots of B. juncea plants. This study conducted in solution culture suggests that plant growth regulators can play a significant role in improving phytoextraction of Au.

  6. Lz-0 × Berkeley: a new Arabidopsis recombinant inbred line population for the mapping of complex traits.

    Science.gov (United States)

    Capron, Arnaud; Chang, Xue Feng; Shi, Chun; Beatson, Rodger; Berleth, Thomas

    2014-06-01

    This study describes the generation and test of a genetic resource suited to identify determinants of cell biological traits in plants. The use of quantitative trait loci (QTL) mapping for a better genetic understanding of cell biological traits is still at an early stage, even for biotechnologically important cell properties, such as the dimensions of fiber cells. A common strategy, the mapping of QTLs in recombinant inbred line (RIL) populations, is limited by the fact that the existing RIL populations exploit only a small fraction of the existing natural variation. Here, we report the mapping of QTLs impacting on the length of fiber cells in Arabidopsis inflorescence stems in a newly generated RIL population derived from a cross between the accessions Berkeley and the little known Lz-0. Through inbreeding of individual F(2) plants, a total of 159 new F8 lines were produced and genotyped with a set of 49 single nucleotide polymorphism markers. The population was successfully used not only for the mapping of three QTLs controlling fiber length, but also to map five QTL controlling flowering time under short and long-day conditions. Our study demonstrates the usefulness of this new genetic resource by mapping in it QTLs underlying a poorly explored cellular trait as well as an already better explored regulatory pathway. The new RIL population and an online platform for the continuous supplementation of genetic markers will be generally available to substantially broaden the genetic diversity through which loci with impact on plant quantitative traits can be identified.

  7. Genetic analysis of morphological traits in a new, versatile, rapid-cycling Brassica rapa recombinant inbred line population

    Directory of Open Access Journals (Sweden)

    Hedayat eBagheri

    2012-08-01

    Full Text Available A recombinant inbred line (RIL population was produced based on a wide cross between the rapid-cycling and self-compatible genotypes L58, a Caixin vegetable type, and R-o-18, a yellow sarson oil type. A linkage map based on 160 F7 lines was constructed using 100 SNP, 130 AFLP®, 27 InDel and 13 publicly available SSR markers. The map covers a total length of 1150 cM with an average resolution of 4.3 cM/marker. To demonstrate the versatility of this new population, 17 traits, related to plant architecture and seed characteristics, were subjected to QTL analysis. A total of 47 QTLs were detected, each explaining between 6 to 54% of the total phenotypic variance for the concerned trait. The genetic analysis shows that this population is a useful new tool for analyzing genetic variation for interesting traits in B. rapa, and for further exploitation of the recent availability of the B. rapa whole genome sequence for gene cloning and gene function analysis.

  8. Genetic Differences among the a/J x C57bl/6j Recombinant Inbred Mouse Lines and Their Degree of Association with Glucocorticoid-Induced Cleft Palate

    OpenAIRE

    Liu, Sharon L.; Erickson, Robert P.

    1986-01-01

    Hydrocortisone sodium phosphate was injected intramuscularly into A/J, C57BL/6J and recombinant inbred lines from these two parental lines to study the genetics of steroid-induced cleft palate in a situation of identical maternal and fetal genotypes. The strains were typed for H-2 (the major histocompatibility locus), β-glucuronidase and β2-microglobulin, which served as markers on chromosomes 17, 5 and 2, respectively. Hepatic glucocorticoid binding capacity had been previously measured in H...

  9. Quantitative trait locus analysis of lateral branchrelated traits in cucumber (Cucumis sativus L.) using recombinant inbred lines

    Institute of Scientific and Technical Information of China (English)

    JIANG Su; YUAN XiaoJun; PAN JunSong; HE HuanLe; CAI Run

    2008-01-01

    A group of 224 recombinant inbred lines (RILs) was derived from a narrow cross between 2 cucumber (Cucumis sativus L.) lines, namely, S94 (Northern China type with weak lateral branch growth potential and early lateral branch sprouting time) and S06 (Northern European type with strong lateral branch growth potential and late lateral branch sprouting time). These lines were then used for investigating lateral branch-related traits. A total of 36 quantitative trait loci (QTLs) were detected for the following 4 lateral branch-related traits: lateral branch average length (LBAL), lateral branch total length (LBTL), lateral branch number (LBN), and first lateral branch node (FLBN). Further, each QTL explained 3.1% (Ibtl 2.1, spring) to 32.3% (Ibn2.3, spring) of the observed phenotypic variance. Eleven QTLs (Ibal1.1, Ibtl 1.1, Ibn1.2, fIbn1.2, etc.) for different traits were found to be clustered on the e23m18d-ME23EM6c section (7.4 cM) of linkage group (LG) 1; further, 15 QTLs (Ibal 2.1, Ibtl 2.1, Ibn 2.1, flbn 2.1, etc.)were found to be clustered on the S94A1-ME4SA4a section (13.9 cM) of LG2. Twenty-one QTLs explained more than 10% of the phenotypic variance. Moreover, Ibtl 1.3 (autumn, 26.2%, logarithm of odds (LOD)= 17.4; spring, 26,9%, LOD=17.9) had stable position and contribution in both seasons. Several se-quence-anchor markers (CMBR40, F, CS30, S94A1, CSWTA11B, etc.) were closely linked with some QTLs for LBAL, LBTL, LBN, and FLBN, which can be used for the marker-assisted selection to improve the plant architecture in cucumber breeding.

  10. Quantitative trait locus analysis of lateral branch-related traits in cucumber (Cucumis sativus L.) using recombinant inbred lines

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A group of 224 recombinant inbred lines (RILs) was derived from a narrow cross between 2 cucumber (Cucumis sativus L.) lines, namely, S94 (Northern China type with weak lateral branch growth potential and early lateral branch sprouting time) and S06 (Northern European type with strong lateral branch growth potential and late lateral branch sprouting time). These lines were then used for investigating lateral branch-related traits. A total of 36 quantitative trait loci (QTLs) were detected for the following 4 lateral branch-related traits: lateral branch average length (LBAL), lateral branch total length (LBTL), lateral branch number (LBN), and first lateral branch node (FLBN). Further, each QTL explained 3.1% (lbtl2.1, spring) to 32.3% (lbn2.3, spring) of the observed phenotypic variance. Eleven QTLs (lbal1.1, lbtl1.1, lbn1.2, flbn1.2, etc.) for different traits were found to be clustered on the e23m18d-ME23EM6c section (7.4 cM) of linkage group (LG) 1; further, 15 QTLs (lbal2.1, lbtl2.1, lbn2.1, flbn2.1, etc.) were found to be clustered on the S94A1-ME4SA4a section (13.9 cM) of LG2. Twenty-one QTLs explained more than 10% of the phenotypic variance. Moreover, lbtl1.3 (autumn, 26.2%, logarithm of odds (LOD) = 17.4; spring, 26.9%, LOD = 17.9) had stable position and contribution in both seasons. Several se-quence-anchor markers (CMBR40, F, CS30, S94A1, CSWTA11B, etc.) were closely linked with some QTLs for LBAL, LBTL, LBN, and FLBN, which can be used for the marker-assisted selection to improve the plant architecture in cucumber breeding.

  11. Quantitative trait loci for cell-wall components in recombinant inbred lines of maize (Zea mays L.) I: stalk tissue.

    Science.gov (United States)

    Krakowsky, M D; Lee, M; Coors, J G

    2005-07-01

    Maize silage is a significant energy source for animal production operations, and the efficiency of the conversion of forage into animal mass is an important consideration when selecting cultivars for use as feed. Fiber and lignin are negatively correlated with digestibility of feed, so the development of forage with reduced levels of these cell-wall components (CWCs) is desirable. While variability for fiber and lignin is present in maize germplasm, traditional selection has focused on the yield of the ear rather than the forage quality of the whole plant, and little information is available concerning the genetics of fiber and lignin. The objectives of this study were to map quantitative trait loci (QTLs) for fiber and lignin in the maize stalk and compare them with QTLs from other populations. Stalk samples were harvested from 191 recombinant inbred lines (RILs) of B73 (an inbred line with low-to-intermediate levels of CWCs) x De811 (an inbred line with high levels of CWCs) at two locations in 1998 and one in 1999 and assayed for neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). The QTLs were detected on nine chromosomes, mostly clustered in concordance with the high genetic correlations between NDF and ADF. Adjustment of NDF for ADF and ADF for ADL revealed that most of the variability for CWCs in this population is in ADF. Many of the QTLs detected in this study have also been detected in other populations, and several are linked to candidate genes for cellulose or starch biosynthesis. The genetic information obtained in this study should be useful to breeding efforts aimed at improving the quality of maize silage.

  12. QTL mapping of agronomic waterlogging tolerance using recombinant inbred lines derived from tropical maize (Zea mays L) germplasm.

    Science.gov (United States)

    Zaidi, Pervez Haider; Rashid, Zerka; Vinayan, Madhumal Thayil; Almeida, Gustavo Dias; Phagna, Ramesh Kumar; Babu, Raman

    2015-01-01

    Waterlogging is an important abiotic stress constraint that causes significant yield losses in maize grown throughout south and south-east Asia due to erratic rainfall patterns. The most economic option to offset the damage caused by waterlogging is to genetically incorporate tolerance in cultivars that are grown widely in the target agro-ecologies. We assessed the genetic variation in a population of recombinant inbred lines (RILs) derived from crossing a waterlogging tolerant line (CAWL-46-3-1) to an elite but sensitive line (CML311-2-1-3) and observed significant range of variation for grain yield (GY) under waterlogging stress along with a number of other secondary traits such as brace roots (BR), chlorophyll content (SPAD), % stem and root lodging (S&RL) among the RILs. Significant positive correlation of GY with BR and SPAD and negative correlation with S&RL indicated the potential use of these secondary traits in selection indices under waterlogged conditions. RILs were genotyped with 331 polymorphic single nucleotide polymorphism (SNP) markers using KASP (Kompetitive Allele Specific PCR) Platform. QTL mapping revealed five QTL on chromosomes 1, 3, 5, 7 and 10, which together explained approximately 30% of phenotypic variance for GY based on evaluation of RIL families under waterlogged conditions, with effects ranging from 520 to 640 kg/ha for individual genomic regions. 13 QTL were identified for various secondary traits associated with waterlogging tolerance, each individually explaining from 3 to 14% of phenotypic variance. Of the 22 candidate genes with known functional domains identified within the physical intervals delimited by the flanking markers of the QTL influencing GY and other secondary traits, six have previously been demonstrated to be associated with anaerobic responses in either maize or other model species. A pair of flanking SNP markers has been identified for each of the QTL and high throughput marker assays were developed to facilitate

  13. ROOT TRAITS AND NODULATION OF RECOMBINANT INBRED BEAN LINES FROM A ‘JAMAPA × CALIMA’ POPULATION INOCULATED WITH TWO STRAINS OF RHIZOBIUM

    Science.gov (United States)

    Bean cultivars of Andean and Middle American origin often have contrasting above-ground traits. Less is known, however, of possible differences in root traits of beans from different gene pools. Recombinant inbred lines (RIL) derived from a cross between the Andean cultivar ‘Calima’ and the Middle A...

  14. Randomized trial of recombinant human interleukin-3 versus placebo in prevention of bone marrow depression during first-line chemotherapy for ovarian carcinoma

    NARCIS (Netherlands)

    Hofstra, LS; Kristensen, GB; Willemse, PHB; Vindevoghel, A; Meden, H; Lahousen, M; Oberling, F; Sorbe, B; Crump, M; Sklenar, [No Value; Sluiter, WJ; Kiese, B; Trope, CG; de Vries, EGE

    1998-01-01

    Purpose: To determine whether recombinant human interleukin-3 (rhIL-3) reduces bone marrow depression and improves chemotherapeutic schedule adherence in ovarian cancer patients receiving first-line combination chemotherapy. Patients and Methods: In a randomized multicenter study, 185 patients recei

  15. Genetic mapping of QTLs associated with seed macronutrients accumulation in 'MD96-5722' by 'Spencer' recombinant inbred lines of soybean

    Science.gov (United States)

    Research of genetic mapping of QTLs for macronutrient accumulation in soybean seed is limited. Therefore, the objective of this research was to identify QTLs related to macronutrients (N, C, S, P, K, Ca, and Mg) in seeds in 92 F5:7 recombinant inbred lines developed from a cross between MD 96-5722 (...

  16. Male sterility of triticale lines generated through recombination of triticale and rye maintainers

    Directory of Open Access Journals (Sweden)

    Tomasz Warzecha

    2014-11-01

    Full Text Available The Triticum timopheevi cytoplasmic male sterility (cms system in triticale (xTriticosecale Wittmack suffers from a low frequency of maintainers and environmental instability of the male sterility. On the other hand, the Pampa cms system in rye (Secale cereale exhibits strong male sterility and a low frequency of restorers. Here, we report generating hybrids between maintainers of the T. timopheevi cms system in triticale and maintainers of the rye Pampa cms system. Ten hybrids were obtained. Their hybridity was verified by PCR (polymerase chain reaction using ISSR (inter simple sequence repeats primers. The cms maintaining ability of F2 individuals and their progeny was tested. The F2 plants were crossed to male sterile lines of triticale carrying the T. timopheevi cytoplasm. Among 180 G1 offspring of these crosses, 71 (39.4% were completely male sterile. Fourteen F2 individuals (7.8%, as well as their F2S1 and progeny, generated stable male sterility in G1, G1BC1 and G1BC2 generations after the crosses. Our results suggest that it is possible to produce a more stable cms system in triticale based on the T. timopheevi cytoplasm as compared to the existing one.

  17. Unambiguous identification of recombination lines in single zinc-blende ZnSe nanowires in direct relation to their microstructure.

    Science.gov (United States)

    Saxena, Ankur; Pan, Qi; Ruda, Harry E

    2013-03-15

    We present results on the low-temperature photoluminescence characterization of individual ZnSe nanowires, whose crystal structure was determined to be zinc-blende by transmission electron microscopy on the same individual nanowires as were studied optically. The photoluminescence response from single ZnSe nanowires was found to be dominated by excitonic emission due to native point defects, while no emission peaks related to the unintentional impurities were detected. Two strong photoluminescence lines were observed at 2.785 eV and 2.780 eV, assigned to the excitons bound to deep neutral acceptors related to the vacancies of Zn (V(Zn)) and complexes of V(Zn), respectively. Another recombination peak at 2.800 eV related to the free exciton emission in ZnSe was also observed. Longitudinal optical-phonon replicas of up to three orders were seen for both lines, and the average number of emitted phonons was also determined. The excitonic emission linewidths of 1.5 meV were observed from individual nanowires, which are the narrowest excitonic linewidths reported so far for ZnSe nanowires. The optical response from a single nanowire was also compared to that from a bundle of nanowires, and it was found that the linewidths of excitonic emission from the bundle of nanowires were slightly larger than those from single nanowires, due to the effects of ensemble broadening. It is also suggested that in the case of a bundle of nanowires, the broadening is limited by the nanowire which exhibits the largest excitonic linewidth.

  18. Characterization of an Sf-rhabdovirus-negative Spodoptera frugiperda cell line as an alternative host for recombinant protein production in the baculovirus-insect cell system.

    Science.gov (United States)

    Maghodia, Ajay B; Geisler, Christoph; Jarvis, Donald L

    2016-06-01

    Cell lines derived from the fall armyworm, Spodoptera frugiperda (Sf), are widely used as hosts for recombinant protein production in the baculovirus-insect cell system (BICS). However, it was recently discovered that these cell lines are contaminated with a virus, now known as Sf-rhabdovirus [1]. The detection of this adventitious agent raised a potential safety issue that could adversely impact the BICS as a commercial recombinant protein production platform. Thus, we examined the properties of Sf-RVN, an Sf-rhabdovirus-negative Sf cell line, as a potential alternative host. Nested RT-PCR assays showed Sf-RVN cells had no detectable Sf-rhabdovirus over the course of 60 passages in continuous culture. The general properties of Sf-RVN cells, including their average growth rates, diameters, morphologies, and viabilities after baculovirus infection, were virtually identical to those of Sf9 cells. Baculovirus-infected Sf-RVN and Sf9 cells produced equivalent levels of three recombinant proteins, including an intracellular prokaryotic protein and two secreted eukaryotic glycoproteins, and provided similar N-glycosylation patterns. In fact, except for the absence of Sf-rhabdovirus, the only difference between Sf-RVN and Sf9 cells was SF-RVN produced higher levels of infectious baculovirus progeny. These results show Sf-RVN cells can be used as improved, alternative hosts to circumvent the potential safety hazard associated with the use of Sf-rhabdovirus-contaminated Sf cells for recombinant protein manufacturing with the BICS.

  19. Quantitative trait loci for cell wall components in recombinant inbred lines of maize (Zea mays L.) II: leaf sheath tissue.

    Science.gov (United States)

    Krakowsky, M D; Lee, M; Coors, J G

    2006-02-01

    While maize silage is a significant feed component in animal production operations, little information is available on the genetic bases of fiber and lignin concentrations in maize, which are negatively correlated with digestibility. Fiber is composed largely of cellulose, hemicellulose and lignin, which are the primary components of plant cell walls. Variability for these traits in maize germplasm has been reported, but the sources of the variation and the relationships between these traits in different tissues are not well understood. In this study, 191 recombinant inbred lines of B73 (low-intermediate levels of cell wall components, CWCs) x De811 (high levels of CWCs) were analyzed for quantitative trait loci (QTL) associated with CWCs in the leaf sheath. Samples were harvested from plots at two locations in 1998 and one in 1999 and assayed for neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL). QTL were detected on all ten chromosomes, most in tissue specific clusters in concordance with the high genotypic correlations for CWCs within the same tissue. Adjustment of NDF for its subfraction, ADF, revealed that most of the genetic variation in NDF was probably due to variation in ADF. The low to moderate genotypic correlations for the same CWC across leaf sheath and stalk tissues indicate that some genes for CWCs may only be expressed in certain tissues. Many of the QTL herein were detected in other populations, and some are linked to candidate genes for cell wall carbohydrate biosynthesis.

  20. QTLs conferring cold tolerance at the booting stage of rice using recombinant inbred lines from a japonica x indica cross.

    Science.gov (United States)

    Andaya, V C; Mackill, D J

    2003-04-01

    Low temperature stress is common for rice grown in temperate regions and at high elevations in the tropics. The most senstive stage to this stress is booting, about 11 days before heading. Japonica cultivars are known to be more tolerant than indicas. We constructed a genetic map using 191 recombinant inbred lines derived from a cross between a temperate japonica, M-202, and a tropical indica, IR50, in order to locate quantitative trait loci (QTLs) conferring cold tolerance. The map with a total length of 1,276.8 cM and an average density of one marker every 7.1 cM was developed from 181 loci produced by 175 microsatellite markers. Cold tolerance was measured as the degree of spikelet sterility of treated plants at a 12 degrees C temperature for 5 days in the growth chamber. QTLs on chromosomes 1, 2, 3, 5, 6, 7, 9 and 12 were identified to confer cold tolerance at the booting stage. The QTL contribution to the phenotypic variation ranged from 11 to 17%. The two QTLs with the highest contribution to variation, designated qCTB2a and qCTB3, were derived from the tolerant parent, M-202, each explaining approximately 17% of the phenotypic variance. Two of the eight QTLs for cold tolerance were contributed by IR50.

  1. Use of Recombinant Lines of Wheat for Study of Association Between High Molecular Weight Glutenin Subunits and Flour Quality Characteristics

    Directory of Open Access Journals (Sweden)

    Abdolmajid Rezai

    1997-04-01

    Full Text Available The relation between high molecular weight glutenin subunit alleles and flour quality traits have been studied for 117 recombinant inbred lines (RILs derived by single seed descend to F7 from a cross of Anza, a low quality cultivar, and Inia, a high quality cultivar, containing different alleles at 3 GLU-1 seed storage protein loci. RILs were classified by SDS-PAGE for the 8 possible combinations of the alleles. The allelic variation at the GLU-D1 locus accounted for most of the variation observed in SDS-sedimentation, mixing time and tolerance, and protein content. The GLU-D1 allele encoding the subunits 5+10 was superior to its allelic counterpart, encoding 2+ 12. All three loci had significant single locus additive effects for SDS-sedimentation volume and mixing tolerance. The additive effects of GLU-D1 locus on mixing time and protein content were also significant. The epistatic additive effects were mostly negative and with the exception of αAB and αAD for SDS-sedimentation volume were not significant.

  2. Quantitative trait loci for rice yield-related traits using recombinant inbred lines derived from two diverse cultivars

    Indian Academy of Sciences (India)

    Xu Feng Bai; Li Jun Luo; Wen Hao Yan; Mallikarjuna Rao Kovi; Yong Zhong Xing

    2011-08-01

    The thousand-grain weight and spikelets per panicle directly contribute to rice yield. Heading date and plant height also greatly influence the yield. Dissection of genetic bases of yield-related traits would provide tools for yield improvement. In this study, quantitative trait loci (QTL) mapping for spikelets per panicle, thousand-grain weight, heading date and plant height was performed using recombinant inbred lines derived from a cross between two diverse cultivars, Nanyangzhan and Chuan7. In total, 20 QTLs were identified for four traits. They were located to 11 chromosomes except on chromosome 4. Seven and five QTLs were detected for thousand-grain weight and spikelets per panicle, respectively. Four QTLs were identified for both heading date and plant height. About half the QTLs were commonly detected in both years, 2006 and 2007. Six QTLs are being reported for the first time. Two QTL clusters were identified in regions flanked by RM22065 and RM5720 on chromosome 7 and by RM502 and RM264 on chromosome 8, respectively. The parent, Nanyangzhan with heavy thousand-grain weight, carried alleles with increased effects on all seven thousand-grain weight QTL, which explained why there was no transgressive segregation for thousand-grain weight in the population. In contrast, Chuan7 with more spikelets per panicle carried positive alleles at all five spikelets per panicle QTL except qspp5. Further work on distinction between pleiotropic QTL and linked QTL is needed in two yield-related QTL clusters.

  3. ALMA and VLA Observations of Recombination Lines and Continuum toward the Becklin-Neugebauer object in Orion

    CERN Document Server

    Galvan-Madrid, Roberto; Rodriguez, Luis F

    2012-01-01

    Compared to their centimeter-wavelength counterparts, millimeter recombination lines (RLs) are intrinsically brighter and are free from pressure broadening. We report observations of RLs (H30alpha at 231.9 GHz and H53alpha at 42.9 GHz) and the millimeter and centimeter continuum toward the Becklin-Neugebauer (BN) object in Orion, obtained from the Atacama Large Millimeter/submillimeter Array (ALMA) Science Verification archive and the Very Large Array (VLA). The RL emission appears to be arising from the slowly-moving, dense (N_e=8.4x10^6 cm^-3) base of the ionized envelope around BN. This ionized gas has a relatively low electron temperature (T_e<4900 K) and small (<<10 km s^-1) bulk motions. Comparing our continuum measurements with previous (non)detections, we find evidence that BN could have large flux variations in the mm, but only mild (<30 %) variations in the cm. This could be understood if the free-free continuum of BN arises in an unresolved, unconfined ionized region that preserves its ...

  4. A pilot search for mm-wavelength recombination lines from emerging ionized winds in pre-planetary nebulae candidates

    Science.gov (United States)

    Sánchez Contreras, C.; Báez-Rubio, A.; Alcolea, J.; Bujarrabal, V.; Martín-Pintado, J.

    2017-07-01

    We report the results from a pilot search for radio recombination line (RRL) emission at millimeter wavelengths in a small sample of pre-planetary nebulae (pPNe) and young PNe (yPNe) with emerging central ionized regions. Observations of the H30α, H31α, H39α, H41α, H48β, H49β, H51β, and H55γ lines at 1 and 3 mm have been performed with the IRAM 30 m radio telescope. These lines are excellent probes of the dense inner (≲150 au) and heavily obscured regions of these objects, where the yet unknown agents for PN-shaping originate. We detected mm-RRLs in three objects: CRL 618, MWC 922, and M 2-9. For CRL 618, the only pPN with previous published detections of H41α, H35α, and H30α emission, we find significant changes in the line profiles indicating that current observations are probing regions of the ionized wind with larger expansion velocities and mass-loss rate than 29 yr ago. In the case of MWC 922, we observe a drastic transition from single-peaked profiles at 3 mm (H39α and H41α) to double-peaked profiles at 1 mm (H31α and H30α), which is consistent with maser amplification of the highest frequency lines; the observed line profiles are compatible with rotation and expansion of the ionized gas, probably arranged in a disk+wind system around a 5-10 M⊙ central mass. In M 2-9, the mm-RRL emission appears to be tracing a recent mass outburst by one of the stars of the central binary system. We present the results from non-LTE line and continuum radiative transfer models, which enables us to constrain the structure, kinematics, and physical conditions (electron temperature and density) of the ionized cores of our sample. We find temperatures Te 6000-17 000 K, mean densities ne 105-108 cm-3, radial density gradients ne ∝ r- αn with αn 2-3.5, and motions with velocities of 10-30 km s-1 in the ionized wind regions traced by these mm-wavelength observations. We deduce mass-loss rates of ṀpAGB ≈ 10-6-10-7 M⊙ yr-1, which are significantly higher

  5. Kenaf (Hibiscus cannabinus) and sunn hemp (Crotalaria juncea): Monoculture and polyculture production

    Science.gov (United States)

    Kenaf (Hibiscus cannabinus L.) and sunn hemp (Crotalaria juncea L.) are fast growing summer annual crops with numerous commercial applications (fibers, biofuels, bioremediation, paper pulp, building materials, cover crops, and livestock forages). Field research was conducted in southeast Oklahoma (...

  6. PERTUMBUHAN DAN PRODUKSI TANAMAN SAWI HIJAU Brassica juncea L. PADA BERBAGAI DESAIN HIDROPONIK

    OpenAIRE

    Anjeliza, Rispa Yeusy

    2013-01-01

    The research about growth and production of green mustard Brassica juncea L. on a variety of hydroponic design took place at the Laboratory Division of Biotechnology Activities Research Center, University of Hasanuddin, Makassar and runs from January to March 2013. This research aimed to determine which is more effective hydroponic design to optimize growth and production of green mustard Brassica juncea L. This research was based on a completely randomized design (CRD) with 4 treatments hydr...

  7. Prey to predator size ratio influences foraging efficiency of larval Aeshna juncea dragonflies.

    Science.gov (United States)

    Hirvonen, Heikki; Ranta, Esa

    1996-05-01

    We investigated foraging behaviour of larval dragonflies Aeshna juncea in order to examine the significance of prey density and body size in predator-prey dynamics. A. juncea were offered separately three size-classes of Daphnia magna at low and high densities. The data were collected with direct observations of the foraging individuals. We found that large A. juncea larvae could better enhance their intake of prey biomass as prey size and prey density increased than their smaller conspecifics. However, increasing feeding efficiency of both larval instars was constrained by declining attack success and search rate with increasing prey size and density. With small D. magna, in contrast to large A. juncea, small A. juncea increased their searching efficiency as prey density increased keeping D. magna mortality rate at a constant level. In a predator-prey relationship this indicates stabilizing potential and feeding thresholds set by both prey density and prey-predator size ratio. Attack success dropped with prey size and density, but did not change in the course of the foraging bout. For both A. juncea sizes prey handling times increased as more medium and large prey were eaten. The slope of the increase became steeper with increasing prey-predator size ratio. These observations indicate that components of the predator-prey relationship vary with prey density, contrary to the basic assumptions of functional response equations. Moreover, the results suggest that the effects of prey density change during the ontogeny of predators and prey.

  8. Murine leukemia virus-derived retroviral vector has differential integration patterns in human cell lines used to produce recombinant factor VIII

    Directory of Open Access Journals (Sweden)

    Marcela Cristina Correa de Freitas

    2014-06-01

    Full Text Available OBJECTIVE: Nowadays recombinant factor VIII is produced in murine cells including in Chinese hamster ovary (CHO and baby hamster kidney cells (BHK. Previous studies, using the murine leukemia virus-derived retroviral vector pMFG-FVIII-P140K, modified two recombinant human cell lines, HepG2 and Hek293 to produce recombinant factor VIII. In order to characterize these cells, the present study aimed to analyze the integration pattern of retroviral vector pMFG-FVIII-P140K.METHODS: This study used ligation-mediated polymerase chain reaction to locate the site of viral vector integration by sequencing polymerase chain reaction products. The sequences were compared to genomic databases to characterize respective clones.RESULTS: The retroviral vector presented different and non-random profiles of integration between cells lines. A preference of integration for chromosomes 19, 17 and 11 was observed for HepG2FVIIIdB/P140K and chromosome 9 for Hek293FVIIIdB/P140K. In genomic regions such as CpG islands and transcription factor binding sites, there was no difference in the integration profiles for both cell lines. Integration in intronic regions of encoding protein genes (RefSeq genes was also observed in both cell lines. Twenty percent of integrations occurred at fragile sites in the genome of the HepG2 cell line and 17% in Hek293.CONCLUSION: The results suggest that the cell type can affect the profile of chromosomal integration of the retroviral vector used; these differences may interfere in the level of expression of recombinant proteins.

  9. A functional analysis of N-glycosylation-related genes on sialylation of recombinant erythropoietin in six commonly used mammalian cell lines.

    Science.gov (United States)

    Zhang, Peiqing; Tan, Diana Lifen; Heng, Desmond; Wang, Tianhua; Mariati; Yang, Yuansheng; Song, Zhiwei

    2010-11-01

    Significant efforts have been made to improve the sialylation of recombinant glycoproteins with the aim of extending their in vivo circulation time. Here, we report a systematic functional analysis of 31 N-glycosylation-related genes on sialylation of recombinant EPO in six cell lines. BHK and CHO cells were found to sialylate recombinant EPO most effectively. None of the 31 genes, individually or in combination, was able to improve EPO sialylation in these cells. HEK293, Cos-7 and 3T3 cells showed intermediate sialylation capabilities, whereas NS0 cells sialylated recombinant EPO poorly. Overexpression of ST6GalI, ST3GalIII or ST3GalIV, but not ST3GalVI, was able to improve EPO sialylation in these four cell lines. qRT-PCR experiments revealed that ST3GalIII and ST3GalIV are indeed under expressed in HEK293, 3T3 and NS0 cells. Co-expression of upstream glycogenes failed to synergize with these sialyltransferases to further enhance sialylation, suggesting that the upstream glycogenes are all expressed at sufficient levels.

  10. [Nuclear-cytoplasmic compatibility and the state of mitochondrial and chloroplast DNA regions in alloplasmic recombinant and introgressive lines (H. vulgare)-T. aestivum].

    Science.gov (United States)

    Pershina, L A; Trubacheva, N V; Sinyavskaya, M G; Devyatkina, E P; Kravtsova, L A

    2014-10-01

    Alloplasmic lines combining alien nuclear and cytoplasmic genomes are convenient models for studying the mechanisms of nuclear-cytoplasmic compatibility/incompatibility. In the.present study, we have investigated the correlation between the characters and state of mitochondrial (mt) and chloroplast (cp) DNA regions in alloplasmic recombinant common wheat lines with barley cytoplasm characterized by partial or total fertility. Fertility restoration in the studied lines (Hordeum vulgare)-Triticum aestivum is determined by different ratios of the genetic material of common wheat variety Pyrotrix 28, which is a fertility restorer in the cytoplasm of barley, and varietySaratovskaya 29, which is a fixer of sterility. In partially fertile lines with nuclear genomes dominated by the genetic material of Saratovskaya 29, plant growth and development are suppressed. In these lines we have identified the barley homoplasmy of cpDNA regions infA and rpoB and the heteroplasmy of the 18S/5S mt repeat and the cpDNA ycf5 region. Nuclear-cytoplasmic compatibility in lines with reduced fertility (the genetic material of Pyrotrix 28 predominates in their nuclear genomes) is associated with restoration of normal plant growth and development and the changes in thestate of the studied cpDNA and mtDNA regions towards the wheat type. Thus, in fertile lines, the cpDNA regions (infA, rpoB) and the 18S/5S mt repeat were identified in the homoplasmic wheat state; though the cpDNAycf5 region was in the heteroplasmic state, it was dominated by the wheat type of the copies. The nuclearicytoplasmic compatibility is not broken as a result of introgression of the alien genetic material into the nuclear genome of one of the fertile lines; the plants of introgressive lines are fertile and normally developed, and the states of the cpDNA and mtDNA regions correspond to their states in fertile recombinant lines.

  11. Genome-wide QTL mapping for wheat processing quality parameters in a Gaocheng 8901/Zhoumai 16 recombinant inbred line population

    Directory of Open Access Journals (Sweden)

    Hui Jin

    2016-07-01

    Full Text Available Dough rheological and starch pasting properties play an important role in determining processing quality in bread wheat (Triticum aestivum L.. In the present study, a recombinant inbred line (RIL population derived from a Gaocheng 8901/Zhoumai 16 cross grown in three environments was used to identify quantitative trait loci (QTLs for dough rheological and starch pasting properties evaluated by Mixograph, Rapid Visco-Analyzer (RVA and Mixolab parameters using 90K and 660K single nucleotide polymorphism (SNP chip assays. A high-density linkage map constructed with 46,961 polymorphic SNP markers from the wheat 90K and 660K SNP assays spanned a total length of 4,121 cM, with an average chromosome length of 196.2 cM and marker density of 0.09 cM/marker; 6,596 new SNP markers were anchored to the bread wheat linkage map, with 1,046 and 5,550 markers from the 90K and 660K SNP assays, respectively. Composite interval mapping identified 119 additive QTLs on 20 chromosomes except 4D; among them, 15 accounted for more than 10% of the phenotypic variation across two or three environments. Twelve QTLs for Mixograph parameters, 17 for RVA parameters and 55 for Mixolab parameters were new. Eleven QTL clusters were identified. The closely linked SNP markers can be used in marker-assisted wheat breeding in combination with the Kompetitive Allele Specific PCR (KASP technique for improvement of processing quality in bread wheat.

  12. Reconstruction of linkage maps in the distorted segregation populations of backcross,doubled haploid and recombinant inbred lines

    Institute of Scientific and Technical Information of China (English)

    ZHU ChengSong; WANG FuHua; WANG JianFei; LI GuangJun; ZHANG HongSheng; ZHANG YuanMing

    2007-01-01

    Non-Mendelian segregation of markers,known as distorted segregation,is a common biological phenomenon.Although segregation distortion affects the estimation of map distances and the results of quantitative trait loci(QTL)mapping,the effects of distorted markers are often ignored in the construction of linkage maps and in QTL mapping.Recently,we have developed a multipoint method via a Hidden Markov chain method to reconstruct linkage maps in an F2 population that corrects for bias of map distances between distorted markers.In this article,the method is extended to cover backcross,doubled haploid and recombinant inbred line(RIL)populations.The results from simulated experiments show that:(1)the degree that two linked segregation distortion loci(SDL)affect the estimation of map distances increases as SDL heritability and interval length between adjacent markers increase,whereas sample size has little effect on the bias;(2)two linked SDL result in the underestimation of linkage distances for most cases,overestimation for an additive model with opposite additive effects,and unbiased estimation for an epistatic model with negative additive-by-additive effects;(3)the proposed method can obtain the unbiased estimation of linkage distance.This new method was applied to a rice RIL population with severely distorted segregation to reconstruct the linkage maps,and a bootstrap method was used to Obtain 95% confidence intervals of map distances.The results from real data analysis further demonstrate the utility of our method,which provides a foundation for the inheritance analysis of quantitative and viability traits.

  13. Discrimination against 15N among recombinant inbred lines of Phaseolus vulgaris L. contrasting in phosphorus use efficiency for nitrogen fixation.

    Science.gov (United States)

    Lazali, Mohamed; Bargaz, Adnane; Carlsson, Georg; Ounane, Sidi Mohamed; Drevon, Jean Jacques

    2014-02-15

    Although isotopic discrimination processes during nitrogen (N) transformations influence the outcome of (15)N based quantification of N2 fixation in legumes, little attention has been given to the effects of genotypic variability and environmental constraints such as phosphorus (P) deficiency, on discrimination against (15)N during N2 fixation. In this study, six Phaseolus vulgaris recombinant inbred lines (RILs), i.e. RILs 115, 104, 34 (P deficiency tolerant) and 147, 83, 70 (P deficiency sensitive), were inoculated with Rhizobium tropici CIAT899, and hydroaeroponically grown with P-sufficient (250 μmol P plant(-1) week(-1)) versus P-deficient (75 μmol P plant(-1) week(-1)) supply. Two harvests were done at 15 (before nodule functioning) and 42 (flowering stage) days after transplanting. Nodulation, plant biomass, P and N contents, and the ratios of (15)N over total N content ((15)N/Nt) for shoots, roots and nodules were determined. The results showed lower (15)N/Nt in shoots than in roots, both being much lower than in nodules. P deficiency caused a larger decrease in (15)N/Nt in shoots (-0.18%) than in nodules (-0.11%) for all of the genotypes, and the decrease in shoots was greatest for RILs 34 (-0.33%) and 104 (-0.25%). Nodule (15)N/Nt was significantly related to both the quantity of N2 fixed (R(2)=0.96***) and the P content of nodules (R(2)=0.66*). We conclude that the discrimination against (15)N in the legume N2-fixing symbiosis of common bean with R. tropici CIAT899 is affected by P nutrition and plant genotype, and that the (15)N/Nt in nodules may be used to screen for genotypic variation in P use efficiency for N2 fixation. Copyright © 2013 Elsevier GmbH. All rights reserved.

  14. Mapping Quantitative Trait Loci Associated with Aluminum Toxin Tolerance in NJRIKY Recombinant Inbred Line Population of Soybean(Glycine max)

    Institute of Scientific and Technical Information of China (English)

    Bo Qi; Paul Korir; Tuanjie Zhao; Deyue Yu; Shouyi Chen; Junyi Gai

    2008-01-01

    To investigate the genetic mechanism of AI-tolerance in soybean,a recombinant inbred line population (RIL) with 184 F2:7:11 lines derived from the cross of Kefeng No.1 x Nannong 1138-2 (AI-tolerant x AI-sensitive) were tested in pot experimentwith sand culture medium in net room in Nanjing.Four traits,i.e.plant height,number of leaves,shoot dry weight and root dry weight at seedling stage,were evaluated and used to calculate the average membership index (FAi) as the indicator of AI-tolerance.The composite interval mapping (ClM) under WinQTL Cartographer v.2.5 detected five QTLs (i.e.qFAiol,qFAi-2,qFAi-3,qFAi-4 and qFAi-5),explaining 5.20%-9.07% of the total phenotypic variation individually.While with the multiple interval mapping (MIM) of the same software,five QTLs (qFAi-1,qFAi-5,qFAi-6,qFAi-7,and qFAi-8) explaining 5.7%-24.60% of the total phenotypic variation individually were mapped.Here qFAi-1 and qFAi-5 were detected by both CIM and MIM with the locations in a same flanking marker region,GMKF046-GMKF080 on B1 and satt278-sat_95 on L,respectively.While qFAi-2 under CIM and qFAi-6 under MIM both on D1b2 were located in neighboring regions with their confidence intervals overlapped and might be the same locus.Segregation analysis under major gene plus polygene inheritance model showed that Al-tolerance was controlled by two major genes (h2mg =33.05%) plus polygenes (h2pg=52.73%).Both QTL mapping and segregation analysis confirmed two QTLs responsible for Al-tolerance with relatively low heritability,and there might be a third QTL,confounded with the polygenes in segregation analysis.

  15. Fabrication of an on-line enzyme micro-reactor coupled to liquid chromatography-tandem mass spectrometry for the digestion of recombinant human erythropoietin.

    Science.gov (United States)

    Foo, Hsiao Ching; Smith, Norman W; Stanley, Shawn M R

    2015-04-01

    Our aim was to develop a fast and efficient on-line method using micro-reactors for the digestion and deglycosylation of recombinant human erythropoietin extracted from equine plasma. The trypsin digestion micro reactors were fabricated using fused silica capillaries with either a dextran-modified coating or a porous monolith that was able to immobilise the enzyme. These were both found to be reasonably robust and durable, with the trypsin immobilised on dextran-modified fused silica capillaries offering better reproducibility than the micro-reactor based upon covalent attachment of this enzyme to the polymer. It is also evident that the enzyme attached micro reactors produced some tryptic peptides in a greater yield than in-solution digestion. A peptide-N-glycosidase F reactor was also fabricated and, when coupled with the trypsin reactor, the deaminated peptides T5 DAM and T9 DAM from recombinant human erythropoietin could also be detected by LC-ESI-MS/MS analysis. These results were better than those achieved using off-line digestion plus deglycosylation reactions and the analysis required far less time and effort to complete. The use of this on-line approach improved the sensitivity, efficiency and speed of our confirmation methodology that is based upon detecting the unique peptide segments of recombinant human erythropoietin that has been affinity extracted from positive equine plasma samples.

  16. Target and resistance-related proteins of recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand on myeloma cell lines

    OpenAIRE

    JIAN, YUAN; Chen, Yuling; GENG, CHUANYING; Liu, Nian; YANG, GUANGZHONG; Liu, Jinwei; Li, Xin; Deng, Haiteng; CHEN, WENMING

    2016-01-01

    Recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand (rmhTRAIL) has become a potential therapeutic drug for multiple myeloma (MM). However, the exact targets and resistance mechanisms of rmhTRAIL on MM cells remain to be elucidated. The present study aimed to investigate the target and resistance-related proteins of rmhTRAIL on myeloma cell lines. A TRAIL-sensitive myeloma cell line, RPMI 8226, and a TRAIL-resistance one, U266, were chosen and the differentially ex...

  17. THOR - The HI, OH, Recombination Line Survey of the Milky Way - The pilot study: HI observations of the giant molecular cloud W43

    CERN Document Server

    Bihr, S; Ott, J; Johnston, K G; Brunthaler, A; Anderson, L D; Bigiel, F; Carlhoff, P; Churchwell, E; Glover, S C O; Goldsmith, P F; Heitsch, F; Henning, T; Heyer, M H; Hill, T; Hughes, A; Klessen, R S; Linz, H; Longmore, S N; McClure-Griffiths, N M; Menten, K M; Motte, F; Nguyen-Lu'o'ng, Q; Plume, R; Ragan, S E; Roy, N; Schilke, P; Schneider, N; Smith, R J; Stil, J M; Urquhart, J S; Walsh, A J; Walter, F

    2015-01-01

    To study the atomic, molecular and ionized emission of Giant Molecular Clouds (GMCs), we have initiated a Large Program with the VLA: 'THOR - The HI, OH, Recombination Line survey of the Milky Way'. We map the 21cm HI line, 4 OH lines, 19 H_alpha recombination lines and the continuum from 1 to 2 GHz of a significant fraction of the Milky Way (l=15-67deg, |b|<1deg) at ~20" resolution. In this paper, we focus on the HI emission from the W43 star-formation complex. Classically, the HI 21cm line is treated as optically thin with column densities calculated under this assumption. This might give reasonable results for regions of low-mass star-formation, however, it is not sufficient to describe GMCs. We analyzed strong continuum sources to measure the optical depth, and thus correct the HI 21cm emission for optical depth effects and weak diffuse continuum emission. Hence, we are able to measure the HI mass of W43 more accurately and our analysis reveals a lower limit of M=6.6x10^6 M_sun, which is a factor of 2....

  18. Biogenic Pt uptake and nanoparticle formation in Medicago sativa and Brassica juncea

    Science.gov (United States)

    Bali, Roza; Siegele, Rainer; Harris, Andrew T.

    2010-10-01

    The ability of the facultative metallophyte plants, Medicago sativa ( M. sativa) and Brassica juncea ( B. juncea) to accumulate and translocate platinum (Pt) from aqueous substrates is reported. The influence of Pt concentration in the substrate (5, 10, 20, 40 and 80 ppm), exposure time (24, 48 and 72 h) and substrate pH (2, 3, 5, 7 and 9) was determined. In both plants the concentration of Pt increased with substrate concentration and exposure time. Greater accumulation was detected in the roots of M. sativa than B. juncea, up to a maximum of 94.19 mg Pt g-1 (dry biomass) compared with 38.5 mg Pt g-1 (dry biomass) following exposure to 80 ppm Pt after 72 h exposure, respectively. However, at lower substrate concentrations (5 and 20 ppm) greater quantities of Pt were detected in the shoots of B. juncea, ranging between 0.02 and 0.32 mg Pt g-1 (dry biomass) at 5 ppm across the different time intervals studied, compared with 0.02-0.14 mg Pt g-1 (dry biomass) for M. sativa, suggesting B. juncea to be a better translocator of Pt under idealised conditions at low concentrations. Higher Pt uptake was also observed in acidic media, with a maximum at pH 2 for M. sativa and pH 3 for B. juncea, indicating the role of net surface charge on the bioaccumulation of Pt. Once sequestered Pt(II) was reduced to Pt(0) due to the action of local metabolites. TEM images of M. sativa root samples showed the in vivo formation of Pt nanoparticles between 3 and 100 nm in size and of varying morphologies in the epidermal root cells. In vivo Pt distribution profiles were assessed using proton induced X-ray emission (μ-PIXE) spectroscopy, which showed even distribution across all tissue systems (epidermal, cortical and vascular) within the roots of both M. sativa and B. juncea.

  19. Recombinant λ bacteriophage displaying nanobody towards third domain of HER-2 epitope inhibits proliferation of breast carcinoma SKBR-3 cell line.

    Science.gov (United States)

    Shoae-Hassani, Alireza; Mortazavi-Tabatabaei, Seyed Abdolreza; Sharif, Shiva; Madadi, Shabnam; Rezaei-Khaligh, Hamidreza; Verdi, Javad

    2013-02-01

    Phage display of many nanobodies via filamentous phage in combination with helper phage has been reported by many scientists. The aim of this study was to produce lambda (λ) bacteriophage displaying high-affinity nanobody against HER-2 expressing breast carcinoma cells. Bacteriophage λ is a temperate phage with inherent biological safety in mammalian cells. Here we report the construction of a recombinant λ phage that efficiently expresses specific nanobody towards third domain of HER-2 target on SKBR-3 and MCF-7 cell lines in vitro. We constructed recombinant λ phage particles containing a mammalian expression cassette, C-Myc tagged, encoding VHH gene of camelid anti HER-2 third domain epitope using λ ZAP-cytomegalic virus (CMV) vector. The SKBR-3, MCF-7 and human endometrial stem cells were treated by the nanobody displayed recombinant λ phage. The cell growth inhibition assay was performed by MTT Cell Viability Assay Kit. After the fourth round of biopanning there was a significant enrichment in the phage specifically binding to the antigen. The ratio of targeted phage increased approximately 1,000-fold in the fifth round. The nanobody expressed by λ ZAP-CMV-VHH phagemid cloned in λ bioparticles significantly inhibited the proliferation of HER-2 positive SKBR-3 and MCF-7 cells. Recombinant bacteriophage λ ZAP-CMV-VHH-cDNA could be used efficiently for construction of nanobodies to mortify HER-2 positive breast carcinoma cells as a nanomedical therapeutic.

  20. Pulmonary and hepatic lesions caused by the dehydropyrrolizidine alkaloid-producing plants Crotalaria juncea and Crotalaria retusa in donkeys

    Science.gov (United States)

    The effects and susceptibility of donkeys to Crotalaria juncea and Crotalaria retusa poisoning were determined at high and low doses. Seeds of C. juncea conaining 0.074% of dehyrdropyrrolizidine alkaloids (DHPAs) were administered to three donkeys at 0.3, 0.6 and 1 g/kg body weight daily for 365 day...

  1. Dehydropyrrolizidine alkaloids, including monoesters with an unusual esterifying acid, from cultivated Crotalaria juncea (Sunn Hemp cv. 'Tropic Sun')

    Science.gov (United States)

    Cultivation of Crotalaria juncea L. (Sunn Hemp cv. ‘Tropic Sun’) is recommended as a green manure crop in a rotation cycle to improve soil condition, help control erosion, suppress weeds, and reduce soil nematodes. Because C. juncea belongs to a genus that is known for the production of toxic dehydr...

  2. Subcellular distribution and chemical forms of thorium in Brassica juncea var. foliosa.

    Science.gov (United States)

    Zhou, Sai; Kai, Hailu; Zha, Zhongyong; Fang, Zhendong; Wang, Dingna; Du, Liang; Zhang, Dong; Feng, Xiaojie; Jin, Yongdong; Xia, Chuanqin

    2016-06-01

    Brassica juncea var. foliosa (B. juncea var. foliosa) is a promising species for thorium (Th) phytoextraction due to its large biomass, fast growth rate and high tolerance toward Th. To further understand the mechanisms of Th tolerance, the present study investigated the subcellular distribution and chemical forms of Th found in B. juncea var. foliosa Our results indicated that in both roots and leaves, Th contents in different parts of the cells follow the order of cell wall > membranes and soluble fraction > organelles. In particular, Transmission Electron Microscope (TEM) analysis showed that Th was abundantly located in cell walls of the roots. Additionally, when plants were exposed to different concentrations of Th, we have found that Th existed in B. juncea var. foliosa with different chemical forms. Much of the Th extracted by 2% acetic acid (HAc), 1 M NaCl and HCl in roots with the percentage distribution varied from 47.2% to 62.5%, while in leaves, most of the Th was in the form of residue and the subdominant amount of Th was extracted by HCl, followed by 2% HAc. This suggested that Th compartmentation in cytosol and integration with phosphate or proteins in cell wall might be responsible for the tolerance of B. juncea var. foliosa to the stress of Th.

  3. Growth of Crotalaria juncea L. supplied with mineral nitrogen

    Directory of Open Access Journals (Sweden)

    Elenira Henrique Miranda Mendonça

    2005-03-01

    Full Text Available Plants of Crotalaria juncea inoculated with Rhizobium were treated with nutrient solution containing 10 or 20mg of either N/NO3 or N/NH4.plant-1.week-1 . The control plants received nutrient solution without N. An investigation was conducted on the effect of these sources of N on growth and nitrogen fixation of plants with 30, 60 and 90 days after sowing (DAS. Those that received mineral N presented higher growth than -N plants, but the presence of nodules occurred in all the treatments. Plants treated with NH4 presented higher N content until 60 days. The highest concentrations of leghemoglobin and protein in nodules were found at 30 DAS and there was no difference in leghemoglobin content between treatments for any age and in protein from 60 DAS. Nitrogenase activity did not vary from 60 to 90 days, with the exception of plants that received 20mg N/NO3, where it was higher at 60 daysPlantas de Crotalaria juncea inoculadas com Rhizobium foram tratadas com solução nutritiva contendo 10 ou 20mg de N/NO3 ou N/NH4.planta-1 .semana-1. As plantas controle receberam solução nutritiva sem N. Foi verificado o efeito destas fontes de N no crescimento e fixação de nitrogênio em plantas com 30, 60 e 90 dias após a semeadura (DAS. Aquelas que receberam N mineral apresentaram maior crescimento que plantas -N, mas a presença de nódulos ocorreu em todos os tratamentos. Plantas tratadas com NH4 apresentaram maior conteúdo de N até os 60 dias. As maiores concentrações de leghemoglobina e proteínas em nódulos foram verificadas aos 30 DAS e não houve diferença no conteúdo de leghemoglobina entre os tratamentos, em nenhuma das idades e em proteínas a partir de 60 DAS. A atividade da nitrogenase não variou dos 60 aos 90 dias, com exceção de plantas que receberam 20mg N/NO3, nas quais esta foi maior aos 60 dias.

  4. A novel orf108 co-transcribed with the atpA gene is associated with cytoplasmic male sterility in Brassica juncea carrying Moricandia arvensis cytoplasm.

    Science.gov (United States)

    Ashutosh; Kumar, Pankaj; Dinesh Kumar, V; Sharma, Prakash C; Prakash, Shyam; Bhat, Shripad R

    2008-02-01

    Mitochondrial atpA transcripts were examined in cytoplasmic male sterile (CMS) and fertility restorer lines of CMS (Moricandia arvensis) Brassica juncea. Male sterile flowers had longer atpA transcripts than male fertiles. The mitochondrial atpA region of the CMS line was cloned and sequenced. The 5' and 3' ends of the atpA transcripts of the CMS and the fertility restorer lines were mapped and full-length transcripts were cloned and sequenced. A novel orf108 (open reading frame 108) co-transcribed with the atpA gene was found in the male sterile flowers. In the fertility restorer line, the transcript was cleaved within orf108 to yield monocistronic atpA transcripts.

  5. Inheritance and molecular markers for the seed coat color in Brassica juncea

    Institute of Scientific and Technical Information of China (English)

    Mingli YAN; Zhongsong LIU; Chunyun GUAN; Sheyuan CHEN; Mouzhi YUAN; Xianjun LIU

    2009-01-01

    To elucidate the inheritance of seed coat color in Brassica juncea, Sichuan Yellow inbred (PY) was crossed with the Ziyejie inbred, and their F1 F2 and BC1 and BC2 progenies, derived from backcrossing to PY, were phenotyped for seed coat color. Results showed that the yellow seed coat was controlled by two independent recessive loci. Seven brown-seeded near-isogenic lines were developed by successive backcrosses to PY and by selfing. One of the BC6>F2 populations segregated for a single locus controlling seed coat color was used for mapping. Using the 88 primer pairs from sequence-related amplified polymorphism and the 500 random primers, two markers were found to be linked to the gene for brown seed coat, which were designated as SCM57 and SCM1078. The crossover between these markers and the brown seed coat loci was 2.35% and 7.06%, respectively. A sequence characterized amplified region (SCAR) marker according to Negi et al. (2000), designated as SZ1-331, was found to be linked to the gene for brown seed coat, with a cross-over estimate of 2.35%. The markers were located on the same side of the brown seed coat loci and 2.41, 7.51 and 2.41 cM away from the brown seed coat locus. The seven brown-seeded near-isogenic lines were classified into two groups by three DNA markers. They were located at the same linkage group of the marker RA2-All previously published by Padmaja et al. (2005).

  6. Fontes e níveis de salinidade na germinação de sementes de Crotalaria juncea L. Sources and levels of salinity on germination of Crotalaria juncea L. seeds

    Directory of Open Access Journals (Sweden)

    Anísio da Silva Nunes

    2009-06-01

    Full Text Available Objetivou-se avaliar o efeito dos sais cloreto de cálcio (CaCl2, cloreto de sódio (NaCl e cloreto de potássio (KCl na germinação de Crotalaria juncea L. As sementes foram acondicionadas em caixas tipo gerbox, forradas com duas folhas de papel-filtro e umedecidas com as soluções-teste. Em cada tratamento foram utilizadas 200 sementes, dispostas em quatro repetições de 50 sementes. A indução do estresse salino foi realizada com soluções de NaCl, CaCl2 e KCl com potenciais osmóticos de 0; -0,4; -0,8; -1,2;-1,6 e -2,0 MPa. Foram realizadas avaliações de porcentagem de germinação, fitotoxicidade, tamanho de plântulas, matéria seca e IVG. O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial (3x5+1, composto pelas três fontes salinas em cinco doses e uma solução- teste. O aumento da concentração de KCl provocou toxicidade, prejudicando o IVG, a porcentagem de germinação e o crescimento de plântulas de C. juncea, enquanto as concentrações de NaCl e o CaCl2 não interferiram significativamente na germinação das sementes e no vigor de plântulas.This study was carried out in order to evaluate the effect of calcium chloride (CaCl2, sodium chloride (NaCl, and potassium chloride (KCl salts on germination of Crotalaria juncea L. seeds. The seeds were placed in gerbox boxes lined with two filter paper sheets and wet with test-solution. In each treatment, 200 seeds were used, distributed in four replicates of 50 seeds. The induction of the salt stress was done with solutions of NaCl, CaCl2 and KCl with osmotic potential of 0, -0.4, -0.8, -1.2, -1.6, and -2.0 MPa. Germination percentage, phytotoxicity, seedling size, mass of dry matter, and IVG were assessed. The experimental design was completely randomized, in a (3x5+1 factorial structure, composed of three salt sources in five levels and a test-solution. The increasing concentration of KCl caused toxicity, hindering the IVG, the germination

  7. Hypoglycemic effect of Brassica juncea (seeds) on streptozotocin induced diabetic male albino rat

    Institute of Scientific and Technical Information of China (English)

    T Thirumalai; S Viviyan Therasa; EK Elumalai; E David

    2011-01-01

    Objective: To evaluate the hypoglycemic effect of Brassica juncea (seeds) on streptozotocin induced diabetic male albino rats. Methods: Hypoglycemic activity of Brassica juncea (seeds) aqueous extract at a dose of 250, 350 and 450 mg/kg body weight was evaluated. Adult male Swiss albino rats of six numbers in each group was undertaken for study and evaluated. Results: The serum insulin levels were recorded a significant depletion in all groups, short term as well as long term diabetic animals, when compared to that of normal animals. A significant dosage dependent augmenting effect of the seed extract on the serum insulin was recorded in both short term as well as long term groups. Conclusions: The aqueous seed extract of Brassica juncea has potent hypoglycemic activity in male albino rat.

  8. Complex biallelic IGH rearrangements in IgM-expressing Z-138 cell line : Involvement of downstream immunoglobulin class switch recombination

    NARCIS (Netherlands)

    Guikema, JEJ; Fenton, JAL; de Boer, C; Kleiverda, K; Brink, AATP; Raap, AK; Estrov, Z; Schuuring, E; Kluin, PM

    2005-01-01

    Chromosomal translocations involving the immunoglobulin (Ig) receptor loci usually disrupt and silence these loci. On the basis of observations in follicular lymphoma (FL) with downstream Ig heavy chain (IGH) class switch recombination (CSR), we hypothesized that downstream CSR-mediated chromosomal

  9. Sustained productivity in recombinant Chinese Hamster Ovary (CHO) cell lines: proteome analysis of the molecular basis for a process-related phenotype

    LENUS (Irish Health Repository)

    Meleady, Paula

    2011-07-24

    Abstract Background The ability of mammalian cell lines to sustain cell specific productivity (Qp) over the full duration of bioprocess culture is a highly desirable phenotype, but the molecular basis for sustainable productivity has not been previously investigated in detail. In order to identify proteins that may be associated with a sustained productivity phenotype, we have conducted a proteomic profiling analysis of two matched pairs of monoclonal antibody-producing Chinese hamster ovary (CHO) cell lines that differ in their ability to sustain productivity over a 10 day fed-batch culture. Results Proteomic profiling of inherent differences between the two sets of comparators using 2D-DIGE (Difference Gel Electrophoresis) and LC-MS\\/MS resulted in the identification of 89 distinct differentially expressed proteins. Overlap comparisons between the two sets of cell line pairs identified 12 proteins (AKRIB8, ANXA1, ANXA4, EIF3I, G6PD, HSPA8, HSP90B1, HSPD1, NUDC, PGAM1, RUVBL1 and CNN3) that were differentially expressed in the same direction. Conclusion These proteins may have an important role in sustaining high productivity of recombinant protein over the duration of a fed-batch bioprocess culture. It is possible that many of these proteins could be useful for future approaches to successfully manipulate or engineer CHO cells in order to sustain productivity of recombinant protein.

  10. Characterization of variation and quantitative trait loci related to terpenoid indole alkaloid yield in a recombinant inbred line mapping population of Catharanthus roseus

    Indian Academy of Sciences (India)

    Vishakha Sharma; Swati Chaudhary; Suchi Srivastava; Richa Pandey; Sushil Kumar

    2011-04-01

    Improved Catharanthus roseus cultivars are required for high yields of vinblastine, vindoline and catharanthine and/or serpentine and ajmalicine, the pharmaceutical terpenoid indole alkaloids. An approach to derive them is to map QTL for terpenoid indole alkaloids yields, identify DNA markers tightly linked to the QTL and apply marker assisted selection. Towards the end, 197 recombinant inbred lines from a cross were grown over two seasons to characterize variability for seven biomass and 23 terpenoid indole alkaloids content-traits and yield-traits. The recombinant inbred lines were genotyped for 178 DNA markers which formed a framework genetic map of eight linkage groups (LG), spanning 1786.5 cM, with 10.0 cM average intermarker distance. Estimates of correlations between traits allowed selection of seven relatively more important traits for terpenoid indole alkaloids yields. QTL analysis was performed on them using single marker (regression) analysis, simple interval mapping and composite interval mapping procedures. A total of 20 QTL were detected on five of eight LG, 10 for five traits on LG1, five for four traits on LG2, three for one trait on LG3 and one each for different traits on LG three and four. QTL for the same or different traits were found clustered on three LG. Co-location of two QTL for biomass traits was in accord of correlation between them. The QTL were validated for use in marker assisted selection by the recombinant inbred line which transgressively expressed 16 traits contributory to the yield vinblastine, vindoline and catharanthine from leaves and roots that possessed favourable alleles of 13 relevant QTL.

  11. QTL mapping of starch granule size in common wheat using recombinant inbred lines derived from a PH82-2/Neixiang 188 cross

    OpenAIRE

    Nan Feng; Zhonghu He; Yong Zhang; Xianchun Xia; Yan Zhang

    2013-01-01

    Starch is a crucial component determining the processing quality of wheat (Triticum aestivum L.)-based products. Wheat starch generally contains A-type and B-type starch granules, having different effects on starch properties and end-use qualities. In the present study, 240 recombinant inbred lines (RILs) derived from a PH82-2/Neixiang 188 cross were grown in Anyang, Henan, China, during three cropping seasons. A-type and B-type granule contents were determined using a laser diffraction parti...

  12. Mitochondrial nad2 gene is co-transcripted with CMS-associated orfB gene in cytoplasmic male-sterile stem mustard (Brassica juncea).

    Science.gov (United States)

    Yang, Jing-Hua; Zhang, Ming-Fang; Yu, Jing-Quan

    2009-02-01

    The transcriptional patterns of mitochondrial respiratory related genes were investigated in cytoplasmic male-sterile and fertile maintainer lines of stem mustard, Brassica juncea. There were numerous differences in nad2 (subunit 2 of NADH dehydrogenase) between stem mustard CMS and its maintainer line. One novel open reading frame, hereafter named orfB gene, was located at the downstream of mitochondrial nad2 gene in the CMS. The novel orfB gene had high similarity with YMF19 family protein, orfB in Raphanus sativus, Helianthus annuus, Nicotiana tabacum and Beta vulgaris, orfB-CMS in Daucus carota, atp8 gene in Arabidopsis thaliana, 5' flanking of orf224 in B. napus (nap CMS) and 5' flanking of orf220 gene in CMS Brassica juncea. Three copies probed by specific fragment (amplified by primers of nad2F and nad2R from CMS) were found in the CMS line following Southern blotting digested with HindIII, but only a single copy in its maintainer line. Meanwhile, two transcripts were shown in the CMS line following Northern blotting while only one transcript was detected in the maintainer line, which were probed by specific fragment (amplified by primers of nad2F and nad2R from CMS). Meanwhile, the expression of nad2 gene was reduced in CMS bud compared to that in its maintainer line. We thus suggested that nad2 gene may be co-transcripted with CMS-associated orfB gene in the CMS. In addition, the specific fragment that was amplified by primers of nad2F and nad2R just spanned partial sequences of nad2 gene and orfB gene. Such alterations in the nad2 gene would impact the activity of NADH dehydrogenase, and subsequently signaling, inducing the expression of nuclear genes involved in male sterility in this type of cytoplasmic male sterility.

  13. Replacement of Imu-Cmu intron by NeoR gene alters Imu germ-line expression but has no effect on V(D)J recombination.

    Science.gov (United States)

    Haddad, Dania; Dougier, Hei-Lanne; Laviolette, Nathalie; Puget, Nadine; Khamlichi, Ahmed Amine

    2010-02-01

    The NeoR gene has often been used to unravel the mechanisms underlying long-range interactions between promoters and enhancers during V(D)J assembly and class switch recombination (CSR) in the immunoglobulin heavy chain (IgH) locus. This approach led to the notion that CSR is regulated through competition of germ-line (GL) promoters for activities displayed by the 3' regulatory region (3'RR). This polarized long-range effect of the 3'RR is disturbed upon insertion of NeoR gene in the IgH constant (C(H)) region, where only GL transcription derived from upstream GL promoters is impaired. In the context of V(D)J recombination, replacement of Emu enhancer or Emu core enhancer (cEmu) by NeoR gene fully blocked V(D)J recombination and mu0 GL transcription which originates 5' of DQ52 and severely diminished Imu GL transcription derived from Emu/Imu promoter, suggesting a critical role for cEmu in the regulation of V(D)J recombination and of mu0 and Imu expression. Here we focus on the effect of NeoR gene on mu0 and Imu GL transcription in a mouse line in which the Imu-Cmu intron was replaced by a NeoR gene in the sense-orientation. B cell development was characterized by a marked but incomplete block at the pro-B cell stage. However, V(D)J recombination was unaffected in sorted pro-B and pre-B cells excluding an interference with the accessibility control function of Emu. mu0 GL transcription initiation was relatively normal but the maturation step seemed to be affected most likely through premature termination at NeoR polyadenylation sites. In contrast, Imu transcription initiation was impaired suggesting an interference of NeoR gene with the IgH enhancers that control Imu expression. Surprisingly, in stark contrast with the NeoR effect in the C(H) region, LPS-induced NeoR expression restored Imu transcript levels to normal. The data suggest that Emu enhancer may be the master control element that counteracts the down-regulatory "Neo effect" on Imu expression upon LPS

  14. QTLs for straw quality characteristics identified in recombinant inbred lines of a Hordeum vulgare x H spontaneum cross in a Mediterranean environment

    DEFF Research Database (Denmark)

    Grando, S.; Baum, M.; Ceccarelli, S.;

    2005-01-01

    Barley straw is commonly used as animal feed in many developing countries. Even a small increase in its nutritive value can have a large impact on animal production, and hence, on rural livelihood and human nutrition. Straw quality is strongly affected by environmental factors and is, therefore......, difficult to improve with empirical breeding. The objective of this study was to identify molecular markers to facilitate the improvement of straw quality in barley. For this purpose, we have used the genetic linkage map that was already developed for recombinant inbred lines (RILs) of the cross between...... a Hordeum vulgare cultivar (`Arta') and a H. spontaneum line (H. spontaneum 41-1), covering a total of 890 cM. Straw parameters from RILs grown at Tel Hadya and Breda (ICARDA's research stations) in 2 years (1996/1997 and 1997/1998) were analyzed by NIRS for predicted nutritional characteristics including...

  15. Effectiveness of the Ty-3 Introgression for Conferring Resistance in Recombinant Inbred Lines of Tomato to Bipartite Begomoviruses in Guatemala

    Science.gov (United States)

    Management of begomovirus-incited diseases on tomatoes in Guatemala continues to be a challenge and there continues to be a need to better understand the genetics of resistance to begomoviruses. In this study, the resistant line, Gh13, was crossed with the susceptible line, HUJ-VF, that lacked the ...

  16. Apical Dominance and Planting Density Effects on Weed Suppression by Sunn Hemp (Crotalaria juncea L.)

    Science.gov (United States)

    A field study was conducted in 2008 and 2009 in Citra, Florida to evaluate the effects of seeding rate and removal of apical dominance of sunn hemp (Crotalaria juncea L.) on weed suppression and seed production of sunn hemp. Three seeding rates of sunn hemp were used; a representative seed producti...

  17. Phenotypic characterization of sixteen accessions of sunn hemp (Crotalaria juncea L.)in Florida

    Science.gov (United States)

    Sunn hemp (Crotalaria juncea L.) is a leguminous cover crop that provides benefits to a cropping system including nitrogen accumulation, weed suppression and soil stability. Adoption of sunn hemp as a cover crop is limited primarily due to the availability of seed sources, leading to high seed cost...

  18. PCIB an antiauxin enhances microspore embryogenisis in microspore culture of Brassica juncea

    NARCIS (Netherlands)

    Agarwal, P.K.; Agarwal, P.; Custers, J.B.M.; Liu, C.M.; Bhojwani, S.S.

    2006-01-01

    An efficient protocol to improve microspore embryogenesis is established in an important oleiferous crop, Brassica juncea (Indian mustard). Colchicine was used for enhancing microspore embryogenesis and also to obtain doubled haploid embryos. Colchicine at high concentrations (>10 mg l¿1), for 24

  19. Phosphate-assisted phytoremediation of arsenic by Brassica napus and Brassica juncea: Morphological and physiological response.

    Science.gov (United States)

    Niazi, Nabeel Khan; Bibi, Irshad; Fatimah, Ayesha; Shahid, Muhammad; Javed, Muhammad Tariq; Wang, Hailong; Ok, Yong Sik; Bashir, Safdar; Murtaza, Behzad; Saqib, Zulfiqar Ahmad; Shakoor, Muhammad Bilal

    2017-07-03

    In this study, we examined the potential role of phosphate (P; 0, 50, 100 mg kg(-1)) on growth, gas exchange attributes, and photosynthetic pigments of Brassica napus and Brassica juncea under arsenic (As) stress (0, 25, 50, 75 mg kg(-1)) in a pot experiment. Results revealed that phosphate supplementation (P100) to As-stressed plants significantly increased shoot As concentration, dry biomass yield, and As uptake, in addition to the improved morphological and gas exchange attributes and photosynthetic pigments over P0. However, phosphate-assisted increase in As uptake was substantially (up to two times) greater for B. napus, notably due to higher shoot As concentration and dry biomass yield, compared to B. juncea at the P100 level. While phosphate addition in soil (P100) led to enhanced shoot As concentration in B. juncea, it reduced shoot dry biomass, primarily after 50 and 75 mg kg(-1) As treatments. The translocation factor and bioconcentration factor values of B. napus were higher than B. juncea for all As levels in the presence of phosphate. This study demonstrates that phosphate supplementation has a potential to improve As phytoextraction efficiency, predominantly for B. napus, by minimizing As-induced damage to plant growth, as well as by improving the physiological and photosynthetic attributes.

  20. Biogenic Pt uptake and nanoparticle formation in Medicago sativa and Brassica juncea

    Energy Technology Data Exchange (ETDEWEB)

    Bali, Roza [University of Sydney, Laboratory for Sustainable Technology, School of Chemical and Biomolecular Engineering (Australia); Siegele, Rainer [Australian Nuclear Science and Technology Organization, Institute for Environmental Research (Australia); Harris, Andrew T., E-mail: a.harris@usyd.edu.a [University of Sydney, Laboratory for Sustainable Technology, School of Chemical and Biomolecular Engineering (Australia)

    2010-10-15

    The ability of the facultative metallophyte plants, Medicago sativa (M. sativa) and Brassica juncea (B. juncea) to accumulate and translocate platinum (Pt) from aqueous substrates is reported. The influence of Pt concentration in the substrate (5, 10, 20, 40 and 80 ppm), exposure time (24, 48 and 72 h) and substrate pH (2, 3, 5, 7 and 9) was determined. In both plants the concentration of Pt increased with substrate concentration and exposure time. Greater accumulation was detected in the roots of M. sativa than B. juncea, up to a maximum of 94.19 mg Pt g{sup -1} (dry biomass) compared with 38.5 mg Pt g{sup -1} (dry biomass) following exposure to 80 ppm Pt after 72 h exposure, respectively. However, at lower substrate concentrations (5 and 20 ppm) greater quantities of Pt were detected in the shoots of B. juncea, ranging between 0.02 and 0.32 mg Pt g{sup -1} (dry biomass) at 5 ppm across the different time intervals studied, compared with 0.02-0.14 mg Pt g{sup -1} (dry biomass) for M. sativa, suggesting B. juncea to be a better translocator of Pt under idealised conditions at low concentrations. Higher Pt uptake was also observed in acidic media, with a maximum at pH 2 for M. sativa and pH 3 for B. juncea, indicating the role of net surface charge on the bioaccumulation of Pt. Once sequestered Pt(II) was reduced to Pt(0) due to the action of local metabolites. TEM images of M. sativa root samples showed the in vivo formation of Pt nanoparticles between 3 and 100 nm in size and of varying morphologies in the epidermal root cells. In vivo Pt distribution profiles were assessed using proton induced X-ray emission ({mu}-PIXE) spectroscopy, which showed even distribution across all tissue systems (epidermal, cortical and vascular) within the roots of both M. sativa and B. juncea.

  1. GISH Analysis of Interspecific Hybrid between Brassica juncea and B.carinata%芥菜型油菜与埃塞俄比亚芥杂种基因组原位杂交分析

    Institute of Scientific and Technical Information of China (English)

    杨续蕊; 李勤菲; 葛贤宏; 李再云; 钱伟

    2013-01-01

    Brassica carinata originating from Africa with good adaptation in drought and hot environments is an important genetic source to improve B. juncea, which has been widely sown in the northwest of China. The technique of genomic in situ hybridization( GISH) was adopted to investigate the chromosome segregation of interspecific hybrid (ACBB,2n = 35 ) between B. juncea and B. carinata during meiosis stage. The predominant segregation pattern of chromosomes was 17: 18 in pollen mother cells( PMCs) ,followed by 16: 19, while lagged chromosomes were occasionally observed. And the major segregation pattern of B-genome chromosomes from two species was 8:8. The results suggested that it would be possible to transfer the genetic component of B. carinata into B. juncea via homologous recombination within B genome and homologous recombination among three genomes.%原产于非洲的埃塞俄比亚芥(Brassica carinata,2n=34,BBCC)具有适应于炎热干旱地区种植等特点,是改良我国芥菜型油菜(B.juncea,2n=36,AABB)的重要种质资源.本研究用基因组原位杂交方法(GISH,Genomic in situ hybridization)分析了芥菜型油菜与埃塞俄比亚芥种间杂种花粉母细胞的染色体分离,发现在后期Ⅰ染色体主要以17:18类型分离,其次是16:19,染色体落后现象偶然发生,其中B染色体组以8:8的分离比率较高,表明不同来源的B染色体可正常配对分离.本研究表明,芥菜型油菜与埃塞俄比亚芥远缘杂交,通过染色体同源重组(B染色体间),以及部分同源染色体配对交换的方式(A、B、C基因组间),有可能将埃塞俄比亚芥优良遗传成分转移到芥菜型油菜中.

  2. A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates.

    Science.gov (United States)

    Mochida, Seiya; Tsuzuki, Satoshi; Inouye, Kuniyo; Fushiki, Tohru

    2014-05-01

    Matriptase is an epithelial-derived type-II transmembrane serine protease. This protease is expressed prominently in the villus tip of small-intestinal epithelia at which senescent cells undergo shedding and/or apoptosis. The basement membrane of epithelial cells, including small-intestinal epithelial cells, contains extracellular matrix (ECM) proteins such as fibronectin and laminin. We found previously that high concentrations of a recombinant matriptase catalytic domain (r-MatCD) (e.g. 1 μM) caused an increased detachment of and increases in the activity of apoptotic effector caspase-3 in a rat small-intestinal epithelial IEC-6 line cultured on laminin-coated plates and proposed that at sites with its high level of expression, matriptase contributes to promoting shedding and/or detachment-induced death of epithelial cells through a mechanism mediating loss of cell-ECM adhesion. In this study, we found that even without increasing cell detachment, a high concentration of r-MatCD causes an increase in caspase-3 activity in IEC-6 cells cultured on fibronectin-coated plates, suggesting that the recombinant matriptase can cause apoptosis by a mechanism unrelated to cell detachment. Also, r-MatCD-treated IEC-6 cells on fibronectin were found to display spindle-like morphological changes. We suggest that r-MatCD causes apoptosis of IEC-6 on fibronectin by a mechanism involving the disruption of cell integrity.

  3. Crotalaria (Crotalaria juncea L.) Heavy Metal Uptake in Eastern Hungary

    Science.gov (United States)

    László Phd, M., ,, Dr.

    2009-04-01

    Summary: Soil condition, plant production and ecological protection are most important parts of the sustainable agricultural activity on all over the world nowadays. Soils, their fertility, their content of different macro-, mezo-, micro-, trace elements have almost always dictated the spread of agricultural farmlands, including the plant production-, yield harvest levels and yield element contents possible. The success of agriculturists in the 20th and 21th century, particularly in the Europe has relied on inproved soil fertility managements, appropriate crop production and environmental protection. We can test and improve the situations by using different plant species (Crotalaria juncea L.) x macro nutrients (nitrogen) x chelating agents (Desferal as deferoxamine-mesilate: C25H48N6O8-CH4O3S) methods. Crotalaria has a very potential and important role in soil fertility as a green manure crop in the design of plant rotation to field plant production, in the animal foraging as a fodder-crop with a high protein content (30%) and in the pytoremediation possibilities. Field experiment was carried out on a calcareous chernozem meadow soil (Kunság-region of Hungary, Kunmadaras) in partly of crotalaria experiment series (5 years) in 2001. The agrochemical parameters of the ploughed layer of the region soils were as follows: humus 2.5-3.0%, pH (H2O) 7.7, pH (KCl) 7.0, LE (Lakanen & Erviö 1971 [3])-P2O5 183-218 mg kg-1, LE-K2O 82-115 mg kg-1, LE-Ca 1.3%, LE-Mg 56-60 mg kg-1, LE-Mn 45 mg kg-1 according to soil analysis. Nitrogen (N) x Desferal ("D"-Novartis Pharma AG Basie [7], Switzerland, Suiza 500 mg) x Genotype ("G"-India-University of Agricultural Sciences, Bangalore) x Time (T) experiment involved The N levels were 0, 100, 200 and 300 kg ha-1 year-1, and Desferal 0 and 20 kg ha-1 year-1. The plot size had an area of 4x2=8 m2. Experimental datas were estimated by MANOVA of SPSS. The main results can be summarised as follows: a., At harvest, total air dry phytomass

  4. Characteristics of human cell line, F2N78, for the production of recombinant antibody in fed-batch and perfusion cultures.

    Science.gov (United States)

    Seo, Joon Serk; Min, Byung Sub; Kwon, Young-Bum; Lee, Soo-Young; Cho, Jong-Moon; Park, Keun-Hee; Yang, Yae Ji; Maeng, Ki Eun; Chang, Shin-Jae; Kim, Dong-Il

    2016-03-01

    A human hybrid cell line, F2N78, was developed by somatic fusion of HEK293 and Namalwa cells for the production recombinant biopharmaceutical proteins. In this study, we performed perfusion culture to verify its potential in culture process used for human cell expression platform. Cell viability could be maintained over 90% and high viable cell density was obtained at higher than 1.0 × 10(7) cells/mL by bleeding process in perfusion culture. The cells were adapted well in both culture modes, but there were apparent differences in protein quality. Compared to fed-batch culture, degalactosylated forms such as G0F and G0 as well as Man5 showed no significant increases in perfusion culture. In terms of charge variants, acidic peaks increased, whereas main peaks constantly decreased according to the length of culture period in both methods.

  5. Genetic parameters and selection for resistance to bacterial spot in recombinant F6 lines of Capsicum annuum

    Directory of Open Access Journals (Sweden)

    Messias Gonzaga Pereira

    2009-01-01

    Full Text Available This study aimed to advance generations and select superior sweet pepper genotypes with resistance tobacterial spot, using the breeding method Single Seed Descent (SSD based on the segregating population derived from thecross between Capsicum annuum L. UENF 1421 (susceptible, non-pungent and UENF 1381 (resistant, pungent. Thesegregating F3 generation was grown in pots in a greenhouse until the F5 generation. The F6 generation was grown in fieldconditions. The reaction to bacterial spot was evaluated by inoculation with isolate ENA 4135 of Xanthomonas campestris pv.vesicatoria, based on a score scale and by calculating the area under the disease progress curve (AUDPC. The presence orabsence of capsaicin was also assessed. Eighteen F6 lines were bacterial leaf spot-resistant. Since no capsaicin was detectedin the F6 lines 032, 316, 399, 434, and 517, these will be used in the next steps of the sweet pepper breeding program.

  6. Towards mouse models of perseveration: a heritable component in extinction of operant behavior in fourteen standard and recombinant inbred mouse lines.

    Science.gov (United States)

    Malkki, Hemi A I; Donga, Laura A B; de Groot, Sabine E; Battaglia, Francesco P; Pennartz, Cyriel M A

    2011-09-01

    Extinction of instrumental responses is an essential skill for adaptive behavior such as foraging. So far, only few studies have focused on extinction following appetitive conditioning in mice. We studied extinction of appetitive operant lever-press behavior in six standard inbred mouse strains (A/J, C3H/HeJ, C57BL/6J, DBA/2J, BALB/cByJ and NOD/Ltj) and eight recombinant inbred mouse lines. From the response rates at the end of operant and extinction training we computed an extinction index, with higher values indicating better capability to omit behavioral responding in absence of reward. This index varied highly across the mouse lines tested, and the variability was partially due to a significant heritable component of 12.6%. To further characterize the relationship between operant learning and extinction, we calculated the slope of the time course of extinction across sessions. While many strains showed a considerable capacity to omit responding when lever pressing was no longer rewarded, we found a few lines showing an abnormally high perseveration in lever press behavior, showing no decay in response scores over extinction sessions. No correlation was found between operant and extinction response scores, suggesting that appetitive operant learning and extinction learning are dissociable, a finding in line with previous studies indicating that these forms of learning are dependent on different brain areas. These data shed light on the heritable basis of extinction learning and may help develop animal models of addictive habits and other perseverative disorders, such as compulsive food seeking and eating.

  7. Modelling of spectral lines emitted by hydrogen isotopes for ionising and recombining plasma conditions of tokamak edges

    Energy Technology Data Exchange (ETDEWEB)

    Koubiti, M.; Marandet, Y.; Godbert-Mouret, L.; Stamm, R.; Touati, K. [Universite de Provence, Marseille (France). PIIM; Loch, S.; Summers, H.P. [Strathclyde Univ., Glasgow (United Kingdom). Dept. of Physics and Applied Physics; Meigs, A. [UKAEA-Euratom Fusion Association, Culham Science Centre, Abingdon (United Kingdom); Capes, H.; Escarguel, A.; Guirlet, R.; Michelis, C. de [Association Euratom-CEA, Centre d' Etudes de Cadarache, 13 - Saint-Paul-lez-Durance (France). Dept. de Recherches sur la Fusion Controlee

    2002-07-01

    The plasma in the periphery or in a divertor chamber of a magnetic fusion device plays an important role in the spread of particle and heat power. Optimising the role of such a plasma requires its characterization and the understanding of all the mutual interactions of its constituents (ions, electrons, and neutrals), and also their interactions with the device materials. Among the various diagnostic methods, high-resolution passive spectroscopy is well suitable for such a purpose since it is non-intrusive. However, diagnostics based on this method requires an accurate modelling of the observed spectra. A fitting routine has been developed for the interpretation of D{alpha} spectra measured in front of a neutraliser plate of the Tore-Supra Ergodic Divertor. It takes into account both Zeeman and Doppler effects, the instrumental function, and the relaxation of the neutrals induced by elastic collisions with the plasma ions. On the other side, in order to study high-n Balmer lines and the continuum emission of deuterium in detached divertor plasmas, a more complex lineshape code developed for Stark broadening has been recently coupled to a code accounting for line-merging into the continuum. The code allows the convolution with a Gaussian (or a Lorentzian) to account for the Doppler broadening and the instrumental function. The Stark lineshape code has also been updated to include Zeeman effect and can be used for conditions where both Stark and Zeeman effects are important like in ITER-divertor plasmas. (orig.)

  8. Generation of a tenascin-C-CreER2 knockin mouse line for conditional DNA recombination in renal medullary interstitial cells.

    Directory of Open Access Journals (Sweden)

    Wenjuan He

    Full Text Available Renal medullary interstitial cells (RMIC are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure. Here, we generated a RMIC specific tenascin-C promoter driven inducible CreER2 knockin mouse line with an EGFP reporter. Similar as endogenous tenascin-C expression, the reporter EGFP expression in the tenascin-C-CreER2(+/- mice was observed in the inner medulla of the kidney, and co-localized with COX2 but not with AQP2 or AQP1, suggesting selective expression in RMICs. After recombination (tenascin-C-CreER2(+/-/ROSA26-lacZ(+/- mice + tamoxifen, β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs. Cre activity was not obvious in other major organs or without tamoxifen treatment. This inducible RMIC specific Cre mouse line should therefore provide a novel tool to manipulate genes of interest in RMICs.

  9. Generation and Characterization of a MYF5 Reporter Human iPS Cell Line Using CRISPR/Cas9 Mediated Homologous Recombination.

    Science.gov (United States)

    Wu, Jianbo; Hunt, Samuel D; Xue, Haipeng; Liu, Ying; Darabi, Radbod

    2016-01-05

    Human iPS cells hold great promise for disease modeling and treatment of degenerative disorders including muscular dystrophies. Although a few research groups have used them for skeletal muscle differentiation, most were based on gene over-expression or long-term mesenchymal differentiation and retrospective identification of myogenic cells. Therefore, this study was aimed to generate a knock-in reporter human iPS cell line for MYF5, as an early myogenic specification gene, to allow prospective identification and purification of myogenic progenitors from human iPS cells. By using a CRISPR/Cas9 double nickase strategy, a 2A-GFP reporter was inserted before the stop codon of the MYF5 gene using homologous recombination. This approach allowed for highly efficient in-frame targeting of MYF5 in human iPS cells. Furthermore, in order to prove the reporter function, endogenous MYF5 expression was induced using a novel dead Cas9-VP160 transcriptional activator. Induced clones demonstrated appropriate MYF5-GFP co-expression. Finally, to confirm the differentiation potential, reporter human iPS clones were differentiated through embryoid body method and MYF5-GFP(+) myogenic cells were sorted and characterized. These data provides valuable guidelines for generation of knock-in reporter human iPS cell lines for myogenic genes which can be used for disease modeling, drug screening, gene correction and future in vivo applications.

  10. HIGH DENSITY CULTIVATION OF A RECOMBINANT CD-1 CELL LINE PRODUCING PROUROKINASE USING A BIOSILON MICROCARRIER CULTURE SYSTEM

    Institute of Scientific and Technical Information of China (English)

    肖成祖; 黄子才; 张正光; 叶建新; 高丽华; 郭智霞; 程度胜; 周鹤山; 孔惟惟

    1994-01-01

    CD-1,a genetically-engineered CHO cell line,was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density(over 1 ×107 cells/ml).Prourokinase was stably secreted at about 180IU/106 cells/24h.Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers,then reattach and proliferate on fresh microcarries.This makes it very easy to scale up production.The microcarries could be reused several times without affecting adhesion ,proliferation and prourokinase secretion.With CM-PECC membrane radial flow chromatography and MPG chromatography,the prouroknase in conditioned medium could be purfied to a specific activity of 1×105IU/mg of protein.The purification factor was about 600 fold,and approxiamately 90% of the biological activity was recovered.

  11. THOR: The H i, OH, Recombination line survey of the Milky Way. The pilot study: H i observations of the giant molecular cloud W43

    Science.gov (United States)

    Bihr, S.; Beuther, H.; Ott, J.; Johnston, K. G.; Brunthaler, A.; Anderson, L. D.; Bigiel, F.; Carlhoff, P.; Churchwell, E.; Glover, S. C. O.; Goldsmith, P. F.; Heitsch, F.; Henning, T.; Heyer, M. H.; Hill, T.; Hughes, A.; Klessen, R. S.; Linz, H.; Longmore, S. N.; McClure-Griffiths, N. M.; Menten, K. M.; Motte, F.; Nguyen-Luong, Q.; Plume, R.; Ragan, S. E.; Roy, N.; Schilke, P.; Schneider, N.; Smith, R. J.; Stil, J. M.; Urquhart, J. S.; Walsh, A. J.; Walter, F.

    2015-08-01

    To study the atomic, molecular, and ionized emission of giant molecular clouds (GMCs) in the Milky Way, we initiated a large program with the Karl G. Jansky Very Large Array (VLA): "THOR: The H i, OH, Recombination line survey of the Milky Way". We map the 21 cm H i line, 4 OH lines, up to 19 Hα recombination lines and thecontinuum from 1 to 2 GHz of a significant fraction of the Milky Way (l = 15°-67°, | b | ≤ 1°) at an angular resolution of ~ 20″. Starting in 2012, as a pilot study we mapped 4 square degrees of the GMC associated with the W43 star formation complex. The rest of the THOR survey area was observed during 2013 and 2014. In this paper, we focus on the H i emission from the W43 GMC complex. Classically, the H i 21 cm line is treated as optically thin with properties such as the column density calculated under this assumption. This approach might yield reasonable results for regions of low-mass star formation, however, it is not sufficient to describe GMCs. We analyzed strong continuum sources to measure the optical depth along the line of sight, and thus correct the H i 21 cm emission for optical depth effects and weak diffuse continuum emission. Hence, we are able to measure the H i mass of this region more accurately and our analysis reveals a lower limit for the H i mass of M = 6.6-1.8 × 106 M⊙ (vLSR = 60-120 km s-1), which is a factor of 2.4 larger than the mass estimated with the assumption of optically thin emission. The H i column densities are as high as NH i ~ 150 M⊙ pc-2 ≈ 1.9 × 1022 cm-2, which is an order of magnitude higher than for low-mass star formation regions. This result challenges theoretical models that predict a threshold for the H i column density of ~10 M⊙ pc-2, at which the formation of molecular hydrogen should set in. By assuming an elliptical layered structure for W43, we estimate the particle density profile. For the atomic gas particle density, we find a linear decrease toward the center of W43 with

  12. Ultrastructural observations of the early and late stages of gorgonian coral (Junceella juncea) oocytes.

    Science.gov (United States)

    Tsai, Sujune; Jhuang, Yating; Spikings, Emma; Sung, Ping-Jyun; Lin, Chiahsin

    2014-08-01

    The developmental oogenesis of gorgonian coral was investigated at the histological level. The objective of this study was to examine and improve the understanding of Junceella juncea oogenesis using ultrastructural methods, such as histological sectioning and transmission electron microscopy. At least three types of yolk materials were observed in this study: yolk body, lipid granules and cortical alveoli. Some of the complex yolk materials were encompassed by concentric or arched layers of smooth and rough endoplasmic reticulum and the Golgi complex in early stage oocytes. Different types of vesicles were found in both early and late stage oocytes and some granules could be seen inside the empty vesicles. This may be a possible method for elaborating complex yolk materials. Homogeneous yolks from different types of inclusions were abundant and the autosynthesis of yolk may be a major mechanism in J. juncea oocytes. This is the first report of the ultrastructural observation of oogenesis in gorgonian coral species using transmission electron microscopy. Our study obtained relatively detailed information at the ultrastructural level, and it provides an overview of the oocyte ultrastucture of the gorgonian coral J. juncea.

  13. Antispermatogenic and hormonal effects of Crotalaria juncea Linn.seed extracts in male mice

    Institute of Scientific and Technical Information of China (English)

    B.Vijaykumar; I.Sangamma; A.Sharanabasappa; SaraswatiB.Patil

    2004-01-01

    Aim:To evaluate the antifertility activity of various extracts of Crotalaria juncea seeds in male mice.Methods:Adult male mice were gavaged the petroleum ether,benzene and ethanol extracts of C.juncea seeds,25mg.(100g)-1·day1 for 30 days.On day 31 the animals were sacrificed by cervical dislocation and the testes,epididymis,vas deferens,seminal vesicles,prostate gland,bulbourethral gland and levator ani were dissected out and weighed.The organs were processed for biochemical and histological examination.Results:In petroleum ether,benzene and ethanol extracts treated rats,there was a decrease in the weights of testis and accessory reproductive organs.The diameters of the testis and seminiferous tubules were decreased.Spermatogonia,spermatocytes and spermatids in the testis and the sperm count in cauda epididymis were also decreased. There was a significant reduction in the protein and glycogen contents and an increase in the cholesterol content in the testis,epididymis and vas deferens.Of the 3 extracts,the ethanol extract appeared to be the most potent in antispermatogenic activity.When the ethanol extract was tested in immature male mice,there was an antiandrogenic effect as the weights of accessory organs were reduced.Conclusion:The various extracts of C.juncea seeds arrest spermatogenesis and are likely to have an antiandrogenic activity.(Asian J Androl 2004 Mar,6:67-70)

  14. Brassinosteroids Denigrate the Seasonal Stress through Antioxidant Defense System in Seedlings of Brassica juncea L.

    Directory of Open Access Journals (Sweden)

    Sandeep Kumar

    2014-05-01

    Full Text Available The present work has been undertaken to study the effect of exogenously application of 24-epiBL and 28-homoBL on soluble protein, proline contents and antioxidant defense system of Brassica juncea L. RLM 619 under the influence of seasonal stress. It was observed that 24-epiBL and 28-homoBL treatment enhance the soluble protein, dry weight and shoot length of B. juncea seedlings under seasonal stress. If seeds treated with the different concentrations (10-6, 10-8 and 10-10 M of 24-epiBL and 28-homoBL revealed batter growth, protein and proline contents as compare to untreated seedlings. Similarly the activities of antioxidant enzymes SOD, CAT, APOX, DHAR, PPO and Auxinases were enhanced by the application of different concentration of both brassinosteroids, whereas MDA content was decrease with both brassinosteroids treatments. Then we have concluded that both brassinolides have the seasonal stress ameliorative properties in B. juncea seedlings grown under the influence of seasonal stress. This study culminates to the role of brassinolides as an anti-stress property for protection of plant from various types of stresses.

  15. Target and resistance-related proteins of recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand on myeloma cell lines.

    Science.gov (United States)

    Jian, Yuan; Chen, Yuling; Geng, Chuanying; Liu, Nian; Yang, Guangzhong; Liu, Jinwei; Li, Xin; Deng, Haiteng; Chen, Wenming

    2016-06-01

    Recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand (rmhTRAIL) has become a potential therapeutic drug for multiple myeloma (MM). However, the exact targets and resistance mechanisms of rmhTRAIL on MM cells remain to be elucidated. The present study aimed to investigate the target and resistance-related proteins of rmhTRAIL on myeloma cell lines. A TRAIL-sensitive myeloma cell line, RPMI 8226, and a TRAIL-resistance one, U266, were chosen and the differentially expressed proteins between the two cell lines were analyzed prior and subsequent to rmhTRAIL administration by a liquid chromatography-tandem mass spectrometry method. The results showed that following TRAIL treatment, 6 apoptosis-related proteins, calpain small subunit 1 (CPNS1), peflin (PEF1), B-cell receptor-associated protein 31 (BAP31), apoptosis-associated speck-like protein containing CARD (ASC), BAG family molecular chaperone regulator 2 (BAG2) and chromobox protein homolog 3 (CBX3), were upregulated in RPMI 8226 cells while no change was identified in the U266 cells. Furthermore, small ubiquitin-related modifier 1 and several other ubiquitin proteasome pathway (UPP)-related proteins expressed higher levels in TRAIL-resistant cells U266 compared to the RPMI-8226 cells prior and subsequent to rmhTRAIL treatment. These results suggested that CPNS1, PEF1, BAP31, ASC, BAG2 and CBX3 were possibly target proteins of rmhTRAIL on RPMI 8226 cells, while UPP may have a vital role in mediating TRAIL-resistance in U266 cells.

  16. Susceptibility to AcMNPV and Expression of Recombinant Proteins by a Novel Cell Clone Derived from a Trichoplusia ni QAU-BTI-Tn9-4s Cell Line

    Institute of Scientific and Technical Information of China (English)

    Ming Shan; Shi-ying Zhang; Lei Jiang; Ming Ma; Guo-xun Li

    2011-01-01

    It is well known that Tn5B1-4(commercially known as the High Five)cell line is highly susceptible to baculovirus and provides superior production of recombinant proteins when compared to other insect cell lines.But the characteristics of the cell line do not always remain stable and may change upon continuous passage.Recently an alphanodavirus,named Tn5 Cell Line Virus(or TNCL Virus),was identified in High Five cells in particular. Therefore,we established a new cell line,QB-Tn9-4s,from Trichoplusia ni,which was determined to be free of TNCL virus by RT-PCR analysis. In this paper,we describe the development of a novel cell clone,QB-CL-B,from a low passage QB-Tn9-4s cell line and report its susceptibility to ACMNPV,and the level of recombinant protein production. This cell clone was similar to its parental cells QB-Tn9-4s and Tn5B 1-4 cells in morphology and growth rate;although it also showed approximately the same responses to AcMNPV infection and production of occlusion bodies,there were higher levels of recombinant protein production in comparison to QB-Tn9-4s(parental cells)and High5 cells.

  17. Carbon Recombination Lines toward the Riegel-Crutcher Cloud and other Cold HI Regions in the inner Galaxy

    CERN Document Server

    Roshi, D Anish

    2011-01-01

    We report here, for the first time, the association of low frequency CRRL with \\HI\\ self-absorbing clouds in the inner Galaxy and that the CRRLs from the innermost $\\sim 10^{\\circ}$ of the Galaxy arise in the Riegel-Crutcher (R-C) cloud. The R-C cloud is amongst the most well known of \\HI\\ self-absorbing (HISA) regions located at a distance of about 125 pc in the Galactic centre direction. Taking the R-C cloud as an example, we demonstrate that the physical properties of the HISA can be constrained by combining multi-frequency CRRL and \\HI\\ observations. The derived physical properties of the HISA cloud are used to determine the cooling and heating rates. The dominant cooling process is emission of the \\CII\\ 158 \\mum line whereas dominant heating process in the cloud interior is photoelectric emission. Constraints on the FUV flux (G0 $\\sim$ 4 to 7) falling on the R-C cloud are obtained by assuming thermal balance between the dominant heating and cooling processes. The H$_2$ formation rate per unit volume in t...

  18. Quantitative trait loci analysis of flowering time related traits identified in recombinant inbred lines of cowpea (Vigna unguiculata).

    Science.gov (United States)

    Andargie, Mebeasealassie; Pasquet, Remy S; Muluvi, Geoffrey M; Timko, Michael P

    2013-05-01

    Flowering time is a major adaptive trait in plants and an important selection criterion in the breeding for genetic improvement of crop species. QTLs for the time of flower opening and days to flower were identified in a cross between a short duration domesticated cowpea (Vigna unguiculata (L.) Walp.) variety, 524B, and a relatively long duration wild accession, 219-01. A set of 159 F7 lines was grown under greenhouse conditions and scored for the flowering time associated phenotypes of time of flower opening and days to flower. Using a LOD threshold of 2.0, putative QTLs were identified and placed on a linkage map consisting of 202 SSR markers and four morphological loci. A total of five QTLs related to the time of flower opening were identified, accounting for 8.8%-29.8% of the phenotypic variation. Three QTLs for days to flower were detected, accounting for 5.7%-18.5% of the phenotypic variation. The major QTL of days to flower and time of flower opening were both mapped on linkage group 1. The QTLs identified in this study provide a strong foundation for further validation and fine mapping for developing an efficient way to restrain the gene flow between the cultivated and wild plants.

  19. LINES

    Directory of Open Access Journals (Sweden)

    Minas Bakalchev

    2015-10-01

    Full Text Available The perception of elements in a system often creates their interdependence, interconditionality, and suppression. The lines from a basic geometrical element have become the model of a reductive world based on isolation according to certain criteria such as function, structure, and social organization. Their traces are experienced in the contemporary world as fragments or ruins of a system of domination of an assumed hierarchical unity. How can one release oneself from such dependence or determinism? How can the lines become less “systematic” and forms more autonomous, and less reductive? How is a form released from modernistic determinism on the new controversial ground? How can these elements or forms of representation become forms of action in the present complex world? In this paper, the meaning of lines through the ideas of Le Corbusier, Leonidov, Picasso, and Hitchcock is presented. Spatial research was made through a series of examples arising from the projects of the architectural studio “Residential Transformations”, which was a backbone for mapping the possibilities ranging from playfulness to exactness, as tactics of transformation in the different contexts of the contemporary world.

  20. Brassica juncea tested on urban soils moderately contaminated by lead: Origin of contamination and effect of chelates.

    Science.gov (United States)

    Bouquet, Dorine; Braud, Armelle; Lebeau, Thierry

    2017-05-04

    Urban garden soils are a potential repository of heavy metal pollution, resulting from either anthropogenic or geogenic origin. The efficiency of phytoextraction was compared on two garden soils with the same texture and topsoil Pb concentration (170 mg kg(-1)) but not the same origin: one geogenic, the other anthropogenic. Two varieties of Brassica juncea were tested with citric acid (25 mmol kg(-1)) or ethylenediaminetetraacetic acid (EDTA, 2.5 mmol kg(-1)). Geogenic Pb was shown to be two times less available than anthropogenic Pb, as a result of which the phytoextraction efficiency was reduced by 59%. Pb mobility in the soil was solely enhanced with EDTA, which increased the Pb concentration in shoots of B. juncea by between 14 and 26 times in comparison with the control. The highest Pb concentration in shoots still remained low, however (i.e., 45 mg kg(-1) dry weight). Regardless of the chelates introduced, B. juncea 426308 accumulated roughly twice as much lead as B. juncea 211000, but only for the anthropogenic contaminated soil. Under these conditions, the amount of Pb accumulated by B. juncea (even when assisted by EDTA) was not high enough to envision achieving soil clean-up within a reasonable time frame.

  1. Implication of citrate, malate and histidine in the accumulation and transport of nickel in Mesembryanthemum crystallinum and Brassica juncea.

    Science.gov (United States)

    Amari, Taoufik; Lutts, Stanley; Taamali, Manel; Lucchini, Giorgio; Sacchi, Gian Attilio; Abdelly, Chedly; Ghnaya, Tahar

    2016-04-01

    Citrate, malate and histidine have been involved in many processes including metal tolerance and accumulation in plants. These molecules have been frequently reported to be the potential nickel chelators, which most likely facilitate metal transport through xylem. In this context, we assess here, the relationship between organics acids and histidine content and nickel accumulation in Mesembryanthemum crystallinum and Brassica juncea grown in hydroponic media added with 25, 50 and 100 µM NiCl2. Results showed that M. crystallinum is relatively more tolerant to Ni toxicity than B. juncea. For both species, xylem transport rate of Ni increased with increasing Ni supply. A positive correlation was established between nickel and citrate concentrations in the xylem sap. In the shoot of B. juncea, citric and malic acids concentrations were significantly higher than in the shoot of M. crystallinum. Also, the shoots and roots of B. juncea accumulated much more histidine. In contrast, a higher root citrate concentration was observed in M. crystallinum. These findings suggest a specific involvement of malic and citric acid in Ni translocation and accumulation in M. crystallinum and B. juncea. The high citrate and histidine accumulation especially at 100µM NiCl2, in the roots of M. crystallinum might be among the important factors associated with the tolerance of this halophyte to toxic Ni levels.

  2. Recombinant FSH Compared to Clomiphene Citrate as the First-Line in Ovulation Induction in Polycystic Ovary Syndrome Using Newly Designed Pens: A Randomized Controlled Trial

    Directory of Open Access Journals (Sweden)

    Batool Hossein-Rashidi

    2016-05-01

    Full Text Available Objective: Since there is still controversy regarding the best first-line choice for ovulation induction (OI other than clomiphene citrate (CC in infertile women diagnosed with polycystic ovary syndrome (PCOS, the aim of the present study was to compare recombinant human FSH with CC as the first course of OI in these women.Materials and methods: In this pilot randomized controlled trial, 104 infertile women diagnosed with PCOS were randomized in two groups to receive either CC with the dose of 100mg per day from day 3 of a spontaneous or progestin-induced menstruation for 5 days or rFSH with the starting dose of 50 IU daily {and weekly dose increment of as low as 12.5 IU}, on the day4 of the cycle. They were assessed during a single OI course. The pregnancy rate (PR and live birth rate (LBR were the primary outcomes. The follicular response, endometrial thickness, cancellation of the cycles and ovarian hyper stimulation (OHSS rate were the secondary outcomes.Results: Analyzing data of 96 patients using Chi2 and Fischer’s Exact test (44 in rFSH group and 52 in CC group, both PR and LBR were comparable in the two groups {13.6% vs. 9.6% and 11.4% vs. 9.6% respectively}, with the difference not to be significant (p > 0.05. No cases of OHSS or multiple gestations happened during the treatment course.Conclusion: It seems that rFSH is as efficacious as CC while not with more complications for the first-line OI in infertile women with PCOS. However, due to the limitations of the present study including the small population and the single cycle of treatment, our results did not come out to prove this and more studies with larger study population are needed to compare the cumulative PR and LBR.

  3. QTLs for straw quality characteristics identified in recombinant inbred lines of a Hordeum vulgare x H. spontaneum cross in a Mediterranean environment.

    Science.gov (United States)

    Grando, S; Baum, M; Ceccarelli, S; Goodchild, A; El-Haramein, F Jaby; Jahoor, A; Backes, G

    2005-02-01

    Barley straw is commonly used as animal feed in many developing countries. Even a small increase in its nutritive value can have a large impact on animal production, and hence, on rural livelihood and human nutrition. Straw quality is strongly affected by environmental factors and is, therefore, difficult to improve with empirical breeding. The objective of this study was to identify molecular markers to facilitate the improvement of straw quality in barley. For this purpose, we have used the genetic linkage map that was already developed for recombinant inbred lines (RILs) of the cross between a Hordeum vulgare cultivar ('Arta') and a H. spontaneum line (H. spontaneum 41-1), covering a total of 890 cM. Straw parameters from RILs grown at Tel Hadya and Breda (ICARDA's research stations) in 2 years (1996/1997 and 1997/1998) were analyzed by NIRS for predicted nutritional characteristics including neutral detergent fiber, acid detergent fiber, lignin, digestible organic matter in dry matter, voluntary intake, crude protein, and straw morphology (the percentage of blades, sheaths, and stems). Localization of QTLs was performed using Windows QTL Cartographer, version 2.0. Seventy-three QTLs were identified, the majority of which (17) in the driest of the four environments. Only six QTLs were identified in two environments; in five cases, one of the two was the wettest environment. This is discussed in relation to the possibility of improving straw quality in favorable environments where yields are higher, rather than in dry environments where straw quality is already relatively good.

  4. Development and characterization of recombinant chromosome substitution lines (RCSLs) using Hordeum vulgare subsp. spontaneum as a source of donor alleles in a Hordeum vulgare subsp. vulgare background.

    Science.gov (United States)

    Matus, I; Corey, A; Filichkin, T; Hayes, P M; Vales, M I; Kling, J; Riera-Lizarazu, O; Sato, K; Powell, W; Waugh, R

    2003-12-01

    The ancestor of barley (Hordeum vulgare subsp. spontaneum) may be a source of novel alleles for crop improvement. We developed a set of recombinant chromosome substitution lines (RCSLs) using an accession of H. vulgare subsp. spontaneum (Caesarea 26-24, from Israel) as the donor and Hordeum vulgare subsp. vulgare 'Harrington' (the North American malting quality standard) as the recurrent parent via two backcrosses to the recurrent parent, followed by six generations of selfing. Here we report (i) the genomic architecture of the RCSLs, as inferred by simple sequence repeat (SSR) markers, and (ii) the effects of H. vulgare subsp. spontaneum genome segment introgressions in terms of three classes of phenotypes: inflorescence yield components, malting quality traits, and domestication traits. Significant differences among the RCSLs were detected for all phenotypes measured. The phenotypic effects of the introgressions were assessed using association analysis, and these were referenced to quantitative trait loci (QTL) reported in the literature. Hordeum vulgare subsp. spontaneum, despite its overall inferior phenotype, contributed some favorable alleles for agronomic and malting quality traits. In most cases, the introgression of the ancestral genome resulted in a loss of desirable phenotypes in the cultivated parent. Although disappointing from a plant breeding perspective, this finding may prove to be a useful tool for gene discovery.

  5. Genomic rearrangements at the FRA2H common fragile site frequently involve non-homologous recombination events across LTR and L1(LINE) repeats.

    Science.gov (United States)

    Brueckner, Lena M; Sagulenko, Evgeny; Hess, Elisa M; Zheglo, Diana; Blumrich, Anne; Schwab, Manfred; Savelyeva, Larissa

    2012-08-01

    Common fragile sites (cFSs) are non-random chromosomal regions that are prone to breakage under conditions of replication stress. DNA damage and chromosomal alterations at cFSs appear to be critical events in the development of various human diseases, especially carcinogenesis. Despite the growing interest in understanding the nature of cFS instability, only a few cFSs have been molecularly characterised. In this study, we fine-mapped the location of FRA2H using six-colour fluorescence in situ hybridisation and showed that it is one of the most active cFSs in the human genome. FRA2H encompasses approximately 530 kb of a gene-poor region containing a novel large intergenic non-coding RNA gene (AC097500.2). Using custom-designed array comparative genomic hybridisation, we detected gross and submicroscopic chromosomal rearrangements involving FRA2H in a panel of 54 neuroblastoma, colon and breast cancer cell lines. The genomic alterations frequently involved different classes of long terminal repeats and long interspersed nuclear elements. An analysis of breakpoint junction sequence motifs predominantly revealed signatures of microhomology-mediated non-homologous recombination events. Our data provide insight into the molecular structure of cFSs and sequence motifs affected by their activation in cancer. Identifying cFS sequences will accelerate the search for DNA biomarkers and targets for individualised therapies.

  6. Resistance to the two-spotted spider mite (Tetranychus urticae) by acylsucroses of wild tomato (Solanum pimpinellifolium) trichomes studied in a recombinant inbred line population.

    Science.gov (United States)

    Alba, Juan Manuel; Montserrat, Marta; Fernández-Muñoz, Rafael

    2009-01-01

    Trichome-based host plant resistance is a complex mechanism that could be used in tomato breeding to control arthropod pests. The aims of this work were to evaluate the plant traits (density of trichomes and acylsucrose production) and the functional relationships of these traits with mortality, repellence, and oviposition of Tetranychus urticae Koch (Acari: Tetranychidae). We used a population of recombinant inbred lines (RILs) derived from the cross between the wild tomato, Solanum pimpinellifolium L. 'TO-937', and the cultivated tomato, Solanum lycopersicum L. Multiple regression analyses showed that high acylsucrose content and high type-IV trichome density increased mortality and repellence, and reduced oviposition of T. urticae. Single regression analyses showed that a logistic model best explained the relationship between mortality or repellence and acylsucrose content, whereas a negative-exponential model best described the relationship between oviposition and acylsucrose content. Linear models were the best-fits for the three resistance variables with trichome IV density. Probit analysis was used to estimate acylsucrose effective doses, and revealed that 31 and 10% of the RILs produced acylsucrose above the effective doses for 90% mortality or repellence, respectively. Altogether, these results indicate that S. pimpinellifolium may be a suitable genetic source of resistance to spider mites to be used in cultivated tomato.

  7. Effect of gliadins and HMW and LMW subunits of glutenin on dough properties in the F6 recombinant inbred lines from a bread wheat cross.

    Science.gov (United States)

    Nieto-Taladriz, M T; Perretant, M R; Rousset, M

    1994-04-01

    The storage proteins of 64 F2-derived F6 recombinant inbred lines (RILs) from the bread wheat cross 'Prinqual'/'Marengo' were analyzed. Parents differed at four loci: Gli-B1 (coding for gliadins), Glu-B1 (coding for HMW glutenin subunits), Glu-A3/Gli-A1 (coding for LMW glutenin subunits/gliadins) and Glu-D3 (coding for LMW glutenin subunits). The effect of allelic variation at these loci on tenacity, extensibility and dough strength as measured by the Chopin alveograph was determined. Allelic differences at the Glu-B1 locus had a significant effect on only tenacity. None of the allelic differences at either the Glu-A3/Gli-A1 or Glu-D3 loci had a significant effect on quality criteria. Allelic variation at the Gli-B1 locus significantly affected all of the dough properties. Epistatic effects between some of the loci considered contributed significantly to the variation in dough quality. Additive and epistatic effects each accounted for 15% of the variation in tenacity. Epistasis accounted for 15% of the variation in extensibility, whereas additive effects accounted for 4%. Epistasis accounted for 14% of the variation in dough strength, and additivity for 9%. The relative importance of epistatic effects suggest that they should be included in predictive models when breeding for breadmaking quality.

  8. QTL mapping of starch granule size in common wheat using recombinant inbred lines derived from a PH82-2/Neixiang188 cross

    Institute of Scientific and Technical Information of China (English)

    Nan; Feng; Zhonghu; He; Yong; Zhang; Xianchun; Xia; Yan; Zhang

    2013-01-01

    Starch is a crucial component determining the processing quality of wheat(Triticum aestivum L.)-based products. Wheat starch generally contains A-type and B-type starch granules, having different effects on starch properties and end-use qualities. In the present study, 240 recombinant inbred lines(RILs) derived from a PH82-2/Neixiang 188 cross were grown in Anyang, Henan, China, during three cropping seasons. A-type and B-type granule contents were determined using a laser diffraction particle size analyzer, defined as the percentage of total starch volume. A total of 195 SSR and STS markers were used to construct a genetic map. QTL analysis was performed by composite interval mapping. Three QTL for A-type starch granule content were mapped on chromosomes 1DL, 7BL and 4AL, explaining5.6%, 5.2% and 3.8% of the phenotypic variation, respectively. These results provide useful information for improving starch quality in common wheat.

  9. Characterization of opaque2 modifier QTLs and candidate genes in recombinant inbred lines derived from the K0326Y quality protein maize inbred

    KAUST Repository

    Holding, David R.

    2010-11-13

    Quality protein maize (QPM) is a high lysine-containing corn that is based on genetic modification of the opaque2 (o2) mutant. In QPM, modifier genes convert the starchy endosperm of o2 to the vitreous phenotype of wild type maize. There are multiple, unlinked o2 modifier loci (Opm) in QPM and their nature and mode of action are unknown. We previously identified seven Opm QTLs and characterized 16 genes that are differentially up-regulated at a significant level in K0326Y QPM, compared to the starchy endosperm mutant W64Ao2. In order to further characterize these Opm QTLs and the genes up-regulated in K0326Y QPM, we created a population of 314 recombinant inbred lines (RILs) from a cross between K0326Y QPM and W64Ao2. The RILs were characterized for three traits associated with endosperm texture: vitreousness, density and hardness. Genetic linkage analysis of the RIL population confirmed three of the previously identified QTLs associated with o2 endosperm modification in K0326Y QPM. Many of the genes up-regulated in K0326Y QPM showed substantially higher levels of expression in vitreous compared with opaque RILs. These included genes associated with the upstream regulation of the ethylene response pathway, and a gene encoding a regulatory subunit of pyrophosphate-dependent fructose-6-phosphate 1-phosphotransferase, an adaptive enzyme of the glycolytic pathway. © 2010 Springer-Verlag.

  10. Comparison of Near Infrared Spectroscopy Models for Determining Protein and Amylose Contents Between Calibration Samples of Recombinant Inbred Lines and Conventional Varieties of Rice

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hong-jiang; WU Jin-hong; LUO Li-jun; LI Ying; YANG Hua; YU Xin-qiao; WANG Xiao-shan; CHEN Liang; MEI Han-wei

    2007-01-01

    The near infrared spectra of 178 recombinant inbred lines (RILs) from the cross of Ⅱ-32B/Yuezaoxian 6 (YZX6) and 511 varieties in rice were acquired. A total of 80 RILs and 96 cultivars were selected as modeling samples by comparing the spectra similarity primarily. Three partial least square (PLS) regression models were developed, based on the RILs (RIL-model), the varieties (Var-model) and their mixture (Mix-model), for protein content (PC) and amylose content (AC),respectively. Cross validation and outer prediction showed that the models were largely influenced by the range and distribution of modeling samples. The regression model of PC based on the cultivars and the model of AC based on RILs had higher coefficient of determination (r2 ≥ 0.9) and lower root mean square error of cross validation (RMSECVs). The disadvantages of RIL samples for PC model and variety samples for AC model were probably caused by the narrow range of variance. Aberrant predictions were obtained for outer sample with PC or AC outside the range or within the distribution gap of modeling samples. The Mix-models gave more reliable prediction as the distribution of RIL and variety modeling samples were complementary to each other.

  11. Two key genomic regions harbour QTLs for salinity tolerance in ICCV 2 × JG 11 derived chickpea (Cicer arietinum L.) recombinant inbred lines.

    Science.gov (United States)

    Pushpavalli, Raju; Krishnamurthy, Laxmanan; Thudi, Mahendar; Gaur, Pooran M; Rao, Mandali V; Siddique, Kadambot H M; Colmer, Timothy D; Turner, Neil C; Varshney, Rajeev K; Vadez, Vincent

    2015-05-22

    Although chickpea (Cicer arietinum L.), an important food legume crop, is sensitive to salinity, considerable variation for salinity tolerance exists in the germplasm. To improve any existing cultivar, it is important to understand the genetic and physiological mechanisms underlying this tolerance. In the present study, 188 recombinant inbred lines (RILs) derived from the cross ICCV 2 × JG 11 were used to assess yield and related traits in a soil with 0 mM NaCl (control) and 80 mM NaCl (salinity) over two consecutive years. Salinity significantly (P chickpea genetic maps showed that these regions conferred salinity tolerance across two other populations and the markers can be deployed for enhancing salinity tolerance in chickpea. Based on the gene ontology annotation, forty eight putative candidate genes responsive to salinity stress were found on CaLG05 (31 genes) and CaLG07 (17 genes) in a distance of 11.1 Mb and 8.2 Mb on chickpea reference genome. Most of the genes were known to be involved in achieving osmoregulation under stress conditions. Identification of putative candidate genes further strengthens the idea of using CaLG05 and CaLG07 genomic regions for marker assisted breeding (MAB). Further fine mapping of these key genomic regions may lead to novel gene identification for salinity stress tolerance in chickpea.

  12. Resistance to Multiple Soil-Borne Pathogens of the Pacific Northwest, USA Is Colocated in a Wheat Recombinant Inbred Line Population

    Directory of Open Access Journals (Sweden)

    Alison L. Thompson

    2017-04-01

    Full Text Available Soil-borne pathogens of the Pacific Northwest decrease yields in both spring and winter wheat. Pathogens of economic importance include Fusarium culmorum, Pratylenchus neglectus, P. thornei, and Rhizoctonia solani AG8. Few options are available to growers to manage these pathogens and reduce yield loss, therefore the focus for breeding programs is on developing resistant wheat cultivars. A recombinant inbred line population, LouAu (MP-7, NSL 511036, was developed to identify quantitative trait loci (QTL associated with resistance to P. neglectus and P. thornei. This same population was later suspected to be resistant to F. culmorum and R. solani AG8. This study confirms partial resistance to F. culmorum and R. solani AG8 is present in this population. Six major and 16 speculative QTL were identified across seven measured traits. Four of the six major QTL were found within the same genomic region of the 5A wheat chromosome suggesting shared gene(s contribute to the resistance. These QTL will be useful in breeding programs looking to incorporate resistance to soil-borne pathogens in wheat cultivars.

  13. Genetic dissection of maize seedling root system architecture traits using an ultra-high density bin-map and a recombinant inbred line population

    Institute of Scientific and Technical Information of China (English)

    Weibin Song; Baobao Wang; Andrew L Hauck; Xiaomei Dong; Jieping Li; Jinsheng Lai

    2016-01-01

    Maize (Zea mays) root system architecture (RSA) mediates the key functions of plant anchorage and acquisition of nutrients and water. In this study, a set of 204 recombinant inbred lines (RILs) was derived from the widely adapted Chinese hybrid ZD958(Zheng58 ? Chang7-2), genotyped by sequencing (GBS) and evaluated as seedlings for 24 RSA related traits divided into primary, seminal and total root classes. Significant differences between the means of the parental phenotypes were detected for 18 traits, and extensive transgressive segregation in the RIL population was observed for all traits. Moderate to strong relationships among the traits were discovered. A total of 62 quantitative trait loci (QTL) were identified that individually explained from 1.6% to 11.6% (total root dry weight/total seedling shoot dry weight) of the phenotypic variation. Eighteen, 24 and 20 QTL were identified for primary, seminal and total root classes of traits, respectively. We found hotspots of 5, 3, 4 and 12 QTL in maize chromosome bins 2.06, 3.02-03, 9.02-04, and 9.05-06, respectively, implicating the presence of root gene clusters or pleiotropic effects. These results characterized the phenotypic variation and genetic architecture of seedling RSA in a population derived from a successful maize hybrid.

  14. A 590 kb deletion caused by non-allelic homologous recombination between two LINE-1 elements in a patient with mesomelia-synostosis syndrome.

    Science.gov (United States)

    Kohmoto, Tomohiro; Naruto, Takuya; Watanabe, Miki; Fujita, Yuji; Ujiro, Sae; Okamoto, Nana; Horikawa, Hideaki; Masuda, Kiyoshi; Imoto, Issei

    2017-04-01

    Mesomelia-synostoses syndrome (MSS) is a rare, autosomal-dominant, syndromal osteochondrodysplasia characterized by mesomelic limb shortening, acral synostoses, and multiple congenital malformations due to a non-recurrent deletion at 8q13 that always encompasses two coding-genes, SULF1 and SLCO5A1. To date, five unrelated patients have been reported worldwide, and MMS was previously proposed to not be a genomic disorder associated with deletions recurring from non-allelic homologous recombination (NAHR) in at least two analyzed cases. We conducted targeted gene panel sequencing and subsequent array-based copy number analysis in an 11-year-old undiagnosed Japanese female patient with multiple congenital anomalies that included mesomelic limb shortening and detected a novel 590 Kb deletion at 8q13 encompassing the same gene set as reported previously, resulting in the diagnosis of MSS. Breakpoint sequences of the deleted region in our case demonstrated the first LINE-1s (L1s)-mediated unequal NAHR event utilizing two distant L1 elements as homology substrates in this disease, which may represent a novel causative mechanism of the 8q13 deletion, expanding the range of mechanisms involved in the chromosomal rearrangements responsible for MSS.

  15. Patterns of longevity and fecundity at two temperatures in a set of heat-selected recombinant inbred lines of Drosophila melanogaster.

    Science.gov (United States)

    Sambucetti, P; Loeschcke, V; Norry, F M

    2015-12-01

    Quantitative trait loci (QTL) were mapped for longevity and fecundity at two temperatures, 20 and 30 °C, in two sets of recombinant inbred lines (RIL) highly differing in thermotolerance. Early fecundity (EF) and longevity showed a negative association between temperatures. For instance, longevity was higher and fecundity was lower in the RIL panel showing higher life span at 30 °C. One X-linked QTL (7B3-12E) co-localized for longevity and EF at 20 °C, with one QTL allele showing a positive additive effect on longevity and a negative effect on EF. The across-RIL genetic correlation between longevity and EF was not significant within each temperature, and most QTL that affect life span have no effect on EF at each temperature. EF and longevity can mostly be genetically uncoupled in the thermotolerance-divergent RIL within each temperature as opposed to between temperatures. QTL were mostly temperature specific, although some trait-specific QTL showed possible antagonistic effects between temperatures.

  16. Chrysoperla externa (Neuroptera: Chrysopidae and Utetheisa ornatrix (Lepidoptera: Arctiidae on organically grown Crotalaria juncea (Fabaceae Chrysoperla externa (Neuroptera: Chrysopidae e Utetheisa ornatrix (Lepidoptera: Arctiidae sobre Crotalaria juncea (Fabaceae cultivada organicamente

    Directory of Open Access Journals (Sweden)

    M.A. Costa

    2012-09-01

    Full Text Available Chrysoperla externa (Neuroptera: Chrysopidae larvae can avoid foraging on plants of Crotalaria juncea (Fabaceae after the issuance of floral buds, when the prey of Utetheisa ornatrix (Lepidoptera: Arctiidae incorporate toxic pyrrolizidine alkaloids from this plant. This reduces the predation and favors increasing the number of adults and eggs of this defoliator on crops of this plant. The aim of the present paper was to evaluate some biological and ecological aspects of C. externa and U. ornatrix on the organic crop of C. juncea in the EMBRAPA Maize and Sorghum in Sete Lagoas, Minas Gerais State, Brazil. Chrysoperla externa and U. ornatrix were more abundant in the vegetative and flowering stages of C. juncea, respectively, with caterpillars of this defoliator feeding on leaves and seeds of this plant. The duration of the stages/instars, survival, lifetime fecundity, and oviposition showed that the branches of C. juncea are a suitable food for U. ornatrix. The abundance of adults and larvae of C. externa was lower in the flowering and pods stages of C. juncea, respectively, when the postures of U. ornatrix are present, probably due to the toxicity of the eggs of this prey to this predator. During these stages, C. externa may be reared with alternative hosts, and when the crops of C. juncea are scarce, an artificial diet should be used for rearing this defoliator in the laboratory for biological research and the development of biological control tactics.Larvas de Chrysoperla externa (Neuroptera: Chrysopidae podem evitar o forrageamento sobre plantas de Crotalaria juncea (Fabaceae após a emissão de botões florais, quando presas de Utetheisa ornatrix (Lepidoptera: Arctiidae incorporam alcalóides pirrolizidínicos tóxicos dessa planta. Isso reduz a predação e favorece o aumento do número de adultos e ovos desse desfolhador sobre cultivos dessa planta. O objetivo deste trabalho foi avaliar alguns aspectos biológicos e ecológicos de C

  17. Growth of Brassica juncea under chromium stress: influence of siderophores and indole 3 acetic acid producing rhizosphere bacteria.

    Science.gov (United States)

    Rajkumar, M; Lee, Kui Jae; Lee, Wang Hyu; Banu, J Rajesh

    2005-10-01

    Plant growth promoting rhizobacterial (PGPR) strains A3 and S32 have been shown to promote the growth of Brassica juncea under chromium stress which has been related to the microbial production of siderophores and indole 3 acetic acid (IAA). The aim of the present study is to evaluate the importance of siderophores and IAA producing PGPR on the growth of Brassica juncea under chromium stress. The production of IAA and siderophores were observed in the strains A3 and S32, respectively. Both PGPR strains promote the growth of Brassica juncea under chromium stress. The maximum growth was observed in plants inoculated with siderophores producing strain 32. Both the bacterial inoculum did not influence the uptake of chromium by plants. The present observation showed that PGPR isolates A3 and S32 are capable of protecting the plants against the inhibitory effects of chromium by producing the siderophores and IAA.

  18. Dissociative recombination in aeronomy

    Science.gov (United States)

    Fox, J. L.

    1989-01-01

    The importance of dissociative recombination in planetary aeronomy is summarized, and two examples are discussed. The first is the role of dissociative recombination of N2(+) in the escape of nitrogen from Mars. A previous model is updated to reflect new experimental data on the electronic states of N produced in this process. Second, the intensity of the atomic oxygen green line on the nightside of Venus is modeled. Use is made of theoretical rate coefficients for production of O (1S) in dissociative recombination from different vibrational levels of O2(+).

  19. EFFECTS OF ARSENIC CONCENTRATIONS AND FORMS ON GROWTH AND ARSENIC UPTAKE AND ACCUMULATION BY INDIAN MUSTARD (BRASSICA JUNCEA L. GENOTYPES

    Directory of Open Access Journals (Sweden)

    INDIRA CHATURVEDI

    2006-10-01

    Full Text Available By using two Brassica juncea genotypes (Varuna and DHR-9504 a green house experiment was carried out during crop cycle (2003-2004, at Agricultural Farm, Bilaspur, Chhattisgarh, India. In Indian mustard, arsenic extraction by plants increased signifi cantly with increasing arsenic concentrations in soils. Uptake of arsenite by Indian mustard genotypes was higher than that of arsenate. Stunted growth of the plants was also observed in this study. This experiment clearly demonstrated the existence of genotypical variations in tolerance to As toxicity among Brassica juncea genotypes.

  20. [Effect of rye Secale cereale L. chromosomes 1R and 3R on polyembryony expression in hybrid combinations between (Hordeum vulgare L.)-Triticum aestivum L. alloplasmic recombinant lines and wheat T. aestivum L.-rye S. cereale L. substitution lines].

    Science.gov (United States)

    Pershina, L A; Rakovtseva, T S; Belova, L I; Deviatkina, E P; Silkova, O G; Kravtsova, L A; Shchapova, A I

    2007-07-01

    The effect of rye chromosomes on polyembryony was studied for reciprocal hybrid combinations between (Hordeum vulgare L.)-Triticum aestivum L. alloplasmic recombinant lines and five wheat T. aestivum L. (cultivar Saratovskaya 29)-rye Secale cereale L. (cultivar Onokhoiskaya) substitution lines: IR(1D), 2R(2D), 3R(3B), 5R(5A), and 6R(6A), and for direct hybrid combinations between the [H. marinum ssp. gussoneanum (H. geniculatum All.)]-T. aestivum alloplasmic recombinant line and the wheat-rye substitution lines 1R (1A), 1R (1D), and 3R(3B). Chromosomes 1R and 3R of rye cultivar Onokhoiskaya proved to affect the expression of polyembryony in the hybrid combinations that involved the alloplasmic recombinant lines of common wheat as maternal genotypes. Based on this finding, polyembryony was regarded as a phenotypic expression of nuclear-cytoplasmic interactions where an important role is played by rye chromosomes 1R and 3R and the H. vulgare cytoplasm. Consideration is given to the association between the effect of rye chromosomes 1R and 3R on polyembryony in the [(Hordeum)-T. aestivum x wheat-rye substitution lines] hybrid combinations and their stimulating effect on the development on angrogenic embryoids in isolated anther cultures of the wheat-rye substitution lines.

  1. Accumulation, detoxification, and genotoxicity of heavy metals in Indian mustard (Brassica juncea L.).

    Science.gov (United States)

    Seth, C S; Misra, V; Chauhan, L K S

    2012-01-01

    Plants of Indian mustard (Brassica juncea L.) were exposed to different concentrations (15, 30, 60, 120 microM) of (Cd, Cr, Cu, Pb) for 28 and 56 d for accumulation and detoxification studies. Metal accumulation in roots and shoots were analyzed and it was observed that roots accumulated a significant amount of Cd (1980 microg g(-1) dry weight), Cr (1540 microg g(-1) dry weight), Cu (1995 microg g(-1) dry weight), and Pb (2040 microg g(-1) dry weight) after 56 d of exposure, though in shoot this was 1110, 618, 795, and 409 microg g(-1) dry weight of Cd, Cr, Cu, and Pb, respectively. In order to assess detoxification mechanisms, non-protein thiols (NP-SH), glutathione (GSH) and phytochelatins (PCs) were analyzed in plants. An increase in the quantity of NP-SH (9.55), GSH (8.30), and PCs (1.25) micromol g(-1) FW were found at 15 microM of Cd, however, a gradual decline in quantity was observed from 15 microM of Cd onwards, after 56 d of exposure. For genotoxicity in plants, cytogenetic end-points such as mitotic index (MI), micronucleus formation (MN), mitotic aberrations (MA) and chromosome aberrations (CA) were examined in root meristem cells of B. juncea. Exposure of Cd revealed a significant (P < 0.05) inhibition of MI, induction of MA, CA, and MN in the root tips for 24 h. However, cells examined at 24 h post-exposure showed concentration-wise recovery in all the endpoints. The data revealed that Indian mustard could be used as a potential accumulator of Cd, Cr, Cu, and Pb due to a good tolerance mechanisms provided by combined/concerted action of NP-SH, GSH, and PCs. Also, exposure of Cd can cause genotoxic effects in B. juncea L. through chromosomal mutations, MA, and MN formation.

  2. Expression analysis of chitinase upon challenge inoculation to Alternaria wounding and defense inducers in Brassica juncea

    Directory of Open Access Journals (Sweden)

    Sandhya Rawat

    2017-03-01

    Full Text Available Chitinases are the hydrolytic enzymes which belong to the pathogenesis-related (PR protein family and play an important role not only in plant defense but also in various abiotic stresses. However, only a limited number of chitinase genes have been characterised in B. juncea. In this study, we have characterised B. juncea class IV chitinase gene (accession no EF586206 in response to fungal infection, salicylic acid (SA, jasmonic acid (JA treatments and wounding. Gene expression studies revealed that the transcript levels of Bjchitinase (BjChp gene increases significantly both in local and distal tissues after Alternaria infection. Bjchitinase gene was also induced by jasmonic acid and wounding but moderately by salicylic acid. A 2.5 kb class IV chitinase promoter of this gene was isolated from B. juncea by Genome walking (accession no KF055403.1. In-silico analysis of this promoter revealed a number of conserved cis-regulatory elements related to defense, wounding and signalling molecules like SA, and JA. For validation, chitinase promoter was fused to the GUS gene, and the resultant construct was then introduced into Arabidopsis plants. Histochemical analysis of T2 transgenic Arabidopsis plants showed that higher GUS activity in leaves after fungal infection, wounding and JA treatment but weakly by SA. GUS activity was seen in meristematic tissues, young leaves, seeds and siliques. Finally investigation has led to the identification of a pathogen-inducible, developmentally regulated and organ-specific promoter. Present study revealed that Bjchitinase (BjChp promoter is induced during biotic and environmental stress and it can be used in developing finely tuned transgenics.

  3. Novel, human cell line-derived recombinant factor VIII (Human-cl rhFVIII, Nuwiq(®) ) in children with severe haemophilia A: efficacy, safety and pharmacokinetics.

    Science.gov (United States)

    Klukowska, A; Szczepański, T; Vdovin, V; Knaub, S; Jansen, M; Liesner, R

    2015-09-14

    Nuwiq(®) (Human-cl rhFVIII) is a new-generation recombinant factor VIII (rFVIII) protein, without chemical modification or fusion to any other protein, produced in a human cell line. This prospective, open-label, multinational phase III study assessed the efficacy and safety of Human-cl rhFVIII in 59 previously treated patients (PTPs) with severe haemophilia A aged 2-12 years (2-5 [N = 29]; 6-12 [N = 30]) during standard prophylaxis (≥50 exposure days and ≥6 months). Efficacy in treating breakthrough bleeds and during surgical prophylaxis was also assessed. An initial pharmacokinetic assessment (N = 13 per age subgroup) demonstrated comparable results with the one-stage and chromogenic assays. Mean (SD) half-life was 11.9 (5.4) and 13.1 (2.6) hours in children aged 2-5 years and 6-12 years respectively (one-stage assay). Prophylactic efficacy, based on mean monthly bleeding rate, was 'excellent' or 'good' in 91.5% of children for all bleeds and in 96.6% of children for spontaneous bleeds. Mean (SD) annualized bleeding rate was 4.12 (5.22) [median 1.9] for all bleeds, 1.50 (3.32) [median 0] for spontaneous bleeds and 2.34 (3.54) [median 1.57] for traumatic bleeds. There were no major, life-threatening bleeds. Efficacy was 'excellent' or 'good' in the treatment of 82.4% of breakthrough bleeds. Overall efficacy during five major surgeries was rated as 'excellent'. There were no FVIII inhibitors or treatment-related serious adverse events. These results in paediatric PTPs indicate that Human-cl rhFVIII is effective for the prevention and treatment of bleeds. © 2015 The Authors. Haemophilia Published by John Wiley & Sons Ltd.

  4. Production of proinflammatory cytokines in the human THP-1 monocyte cell line following induction by Tp0751,a recombinant protein of Treponema pallidum

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The tissue destruction characteristic of syphilis infection may be caused by inflammation due to Treponema pallidum and the ensuing immune responses to the pathogen.T.pallidum membrane proteins are thought to be potent inducers of inflammation during the early stages of infection.However,the actual membrane proteins that induce inflammatory cytokine production are not known,nor are the molecular mechanisms responsible for triggering and sustaining the inflammatory cascades.In the present study,Tp0751 recombinant protein from T.pallidum was found to induce the production of proinflammatory cytokines,including TNF-α,IL-1βand IL-6,in a THP-1 human monocyte cell line.The signal transduction pathways involved in the production of these cytokines were then further investigated.No inhibition of TNF-a,IL-1β,or IL-6 production was observed following treatment with the SAPK/JNK specific inhibitor SP600125 or with an ERK inhibitor PD98059.By contrast,anti-TLR2 mAb,anti-CD14 mAb,and the p38 inhibitor SB203580 significantly inhibited the production of all three cytokines.In addition,pyrrolidine dithiocarbamate (PDTC),a specific inhibitor of NF-κB,profoundly inhibited the production of these cytokines.Tp0751 treatment strongly activated NF-κB,as revealed by Western blotting.However,NF-κB translocation was significantly inhibited by treatment with PDTC.These results indicated that TLR2,CD14,MAPKs/p38,and NF-κB might be implicated in the inflammatory reaction caused by T.pallidum infection.

  5. Identification of quantitative trait loci controlling root and shoot traits associated with drought tolerance in a lentil (Lens culinaris Medik. recombinant inbred line population

    Directory of Open Access Journals (Sweden)

    Omar Idrissi

    2016-08-01

    Full Text Available Drought is one of the major abiotic stresses limiting lentil productivity in rainfed production systems. Specific rooting patterns can be associated with drought avoidance mechanisms that can be used in lentil breeding programs. In all, 252 co-dominant and dominant markers were used for Quantitative Trait Loci (QTL analysis on 132 lentil recombinant inbred lines based on greenhouse experiments for root and shoot traits during two seasons under progressive drought-stressed conditions. Eighteen QTLs controlling a total of 14 root and shoot traits were identified. A QTL-hotspot genomic region related to a number of root and shoot characteristics associated with drought tolerance such as dry root biomass, root surface area, lateral root number, dry shoot biomass and shoot length was identified. Interestingly, a QTL related to root-shoot ratio, an important trait for drought avoidance, explaining the highest phenotypic variance of 27.6 % and 28.9 % for the two consecutive seasons, respectively, was detected. This QTL was closed to the co-dominant SNP marker TP6337 and also flanked by the two SNP TP518 and TP1280. An important QTL related to lateral root number was found close to TP3371 and flanked by TP5093 and TP6072 SNP markers. Also, a QTL associated with specific root length was identified close to TP1873 and flanked by F7XEM6b SRAP marker and TP1035 SNP marker. These two QTLs were detected in both seasons. Our results could be used for marker-assisted selection in lentil breeding programs targeting root and shoot characteristics conferring drought avoidance as an efficient alternative to slow and labour-intensive conventional breeding methods.

  6. Mapping QTLs for Salt Tolerance in Rice (Oryza sativa L.) by Bulked Segregant Analysis of Recombinant Inbred Lines Using 50K SNP Chip.

    Science.gov (United States)

    Tiwari, Sushma; Sl, Krishnamurthy; Kumar, Vinod; Singh, Balwant; Rao, A R; Mithra Sv, Amitha; Rai, Vandna; Singh, Ashok K; Singh, Nagendra K

    2016-01-01

    Soil salinity is a major constraint to rice production in large inland and coastal areas around the world. Modern high yielding rice varieties are particularly sensitive to high salt stress. There are salt tolerant landraces and traditional varieties of rice but with limited information on genomic regions (QTLs) and genes responsible for their tolerance. Here we describe a method for rapid identification of QTLs for reproductive stage salt tolerance in rice using bulked segregant analysis (BSA) of bi-parental recombinant inbred lines (RIL). The number of RILs required for the creation of two bulks with extreme phenotypes was optimized to be thirty each. The parents and bulks were genotyped using a 50K SNP chip to identify genomic regions showing homogeneity for contrasting alleles of polymorphic SNPs in the two bulks. The method was applied to 'CSR11/MI48' RILs segregating for reproductive stage salt tolerance. Genotyping of the parents and RIL bulks, made on the basis of salt sensitivity index for grain yield, revealed 6,068 polymorphic SNPs and 21 QTL regions showing homogeneity of contrasting alleles in the two bulks. The method was validated further with 'CSR27/MI48' RILs used earlier for mapping salt tolerance QTLs using low-density SSR markers. BSA with 50K SNP chip revealed 5,021 polymorphic loci and 34 QTL regions. This not only confirmed the location of previously mapped QTLs but also identified several new QTLs, and provided a rapid way to scan the whole genome for mapping QTLs for complex agronomic traits in rice.

  7. Mapping QTLs for Salt Tolerance in Rice (Oryza sativa L.) by Bulked Segregant Analysis of Recombinant Inbred Lines Using 50K SNP Chip

    Science.gov (United States)

    Kumar, Vinod; Singh, Balwant; Rao, AR; Mithra SV, Amitha; Singh, Ashok K.; Singh, Nagendra K.

    2016-01-01

    Soil salinity is a major constraint to rice production in large inland and coastal areas around the world. Modern high yielding rice varieties are particularly sensitive to high salt stress. There are salt tolerant landraces and traditional varieties of rice but with limited information on genomic regions (QTLs) and genes responsible for their tolerance. Here we describe a method for rapid identification of QTLs for reproductive stage salt tolerance in rice using bulked segregant analysis (BSA) of bi-parental recombinant inbred lines (RIL). The number of RILs required for the creation of two bulks with extreme phenotypes was optimized to be thirty each. The parents and bulks were genotyped using a 50K SNP chip to identify genomic regions showing homogeneity for contrasting alleles of polymorphic SNPs in the two bulks. The method was applied to ‘CSR11/MI48’ RILs segregating for reproductive stage salt tolerance. Genotyping of the parents and RIL bulks, made on the basis of salt sensitivity index for grain yield, revealed 6,068 polymorphic SNPs and 21 QTL regions showing homogeneity of contrasting alleles in the two bulks. The method was validated further with ‘CSR27/MI48’ RILs used earlier for mapping salt tolerance QTLs using low-density SSR markers. BSA with 50K SNP chip revealed 5,021 polymorphic loci and 34 QTL regions. This not only confirmed the location of previously mapped QTLs but also identified several new QTLs, and provided a rapid way to scan the whole genome for mapping QTLs for complex agronomic traits in rice. PMID:27077373

  8. Identification and validation of quantitative trait loci for seed yield, oil and protein contents in two recombinant inbred line populations of soybean.

    Science.gov (United States)

    Wang, Xianzhi; Jiang, Guo-Liang; Green, Marci; Scott, Roy A; Song, Qijian; Hyten, David L; Cregan, Perry B

    2014-10-01

    Soybean seeds contain high levels of oil and protein, and are the important sources of vegetable oil and plant protein for human consumption and livestock feed. Increased seed yield, oil and protein contents are the main objectives of soybean breeding. The objectives of this study were to identify and validate quantitative trait loci (QTLs) associated with seed yield, oil and protein contents in two recombinant inbred line populations, and to evaluate the consistency of QTLs across different environments, studies and genetic backgrounds. Both the mapping population (SD02-4-59 × A02-381100) and validation population (SD02-911 × SD00-1501) were phenotyped for the three traits in multiple environments. Genetic analysis indicated that oil and protein contents showed high heritabilities while yield exhibited a lower heritability in both populations. Based on a linkage map constructed previously with the mapping population and using composite interval mapping and/or interval mapping analysis, 12 QTLs for seed yield, 16 QTLs for oil content and 11 QTLs for protein content were consistently detected in multiple environments and/or the average data over all environments. Of the QTLs detected in the mapping population, five QTLs for seed yield, eight QTLs for oil content and five QTLs for protein content were confirmed in the validation population by single marker analysis in at least one environment and the average data and by ANOVA over all environments. Eight of these validated QTLs were newly identified. Compared with the other studies, seven QTLs for seed yield, eight QTLs for oil content and nine QTLs for protein content further verified the previously reported QTLs. These QTLs will be useful for breeding higher yield and better quality cultivars, and help effectively and efficiently improve yield potential and nutritional quality in soybean.

  9. The Effects of Crude Methanol Seed Extract of Brassica juncea on Haematological Parameters in Wistar Rats

    Directory of Open Access Journals (Sweden)

    S.A. Ufelle

    2011-08-01

    Full Text Available Brassica juncea, (B. juncea also known as Indian Oriental or Brown Mustard is widely used as a medicinal crop, oil seed, condiment and vegetable and much hematological effects have not been recorded on B. juncea. Thirty (30 male wistar rats aged 2-3 months, weighing 100-200 g were obtained and housed at the Animal House of College of Medicine, University of Nigeria Enugu Campus for this study. The rats were divided into five (5 groups of six (6 rats per group, labeled A to E and were acclimatized for two (2 weeks and were fed with commercially available rat feed. They had access to water and feed ad libitum. Groups A to D were orally administered with graded doses of the crude seed extract (50, 100, 200 and 300 mg/kg body weight respectively daily for 30 days while group E which served as control received no extract. Blood samples (2.0 mL were collected from each rat through the retro orbital plexus of the median canthus into K3- EDTA containers for analysis using standard operative procedures. The results obtained, revealed that the mean Hemoglobin (Hb values of groups C and D (C = 12.6±0.57 and D = 11.1±1.4 g/dL were significantly decreased (p<0.05 when compared with control (E = 14.2±0.74 g/dL. The mean Hematocrit (PCV values of groups C and D (C = 0.38±0.02, and D = 0.34±0.04 L/L were also significantly decreased (p<0.05 compared with control (E = 0.43±0.02 L/L. The mean Total WBC values of all the treated groups were significantly increased (A = 6.45±0.3, B = 6.23±0.83, C = 6.0±0.34, and D = 6.05±0.41 x 109/L when compared with control E = 4.28±0.17×109/L (p<0.05. The mean platelet values were significantly decreased in all the treated groups (A = 30.00±4.1, B = 30.00±5.8, C = 46.25±7.5 and D = 33.75±2.5 x 109/L when compared with control E = 86.25±4.8×109/L. This study revealed that crude methanol seed extract of Brassica juncea affects some hematological parameters when orally administered at different concentrations.

  10. Shoot and Root Traits Contribute to Drought Resistance in Recombinant Inbred Lines of MD 23–24 × SEA 5 of Common Bean

    Science.gov (United States)

    Polania, Jose; Rao, Idupulapati M.; Cajiao, Cesar; Grajales, Miguel; Rivera, Mariela; Velasquez, Federico; Raatz, Bodo; Beebe, Stephen E.

    2017-01-01

    Drought is the major abiotic stress factor limiting yield of common bean (Phaseolus vulgaris L.) in smallholder systems in Latin America and eastern and southern Africa; where it is a main source of protein in the daily diet. Identification of shoot and root traits associated with drought resistance contributes to improving the process of designing bean genotypes adapted to drought. Field and greenhouse studies were conducted at the International Center for Tropical Agriculture (CIAT), Palmira, Colombia to determine the relationship between grain yield and different shoot and root traits using a recombinant inbred lines (RILs) population (MD23–24 × SEA 5) of common bean. The main objectives of this study were to identify: (i) specific shoot and root morpho-physiological traits that contribute to improved resistance to drought and that could be useful as selection criteria in breeding beans for drought resistance; and (ii) superior genotypes with desirable shoot and root traits that could serve as parents in breeding programs that are aimed at improving drought resistance. A set of 121 bean genotypes (111 RILs, 2 parents, 8 checks) belonging to the Mesoamerican gene pool and one cowpea variety were evaluated under field conditions with two levels of water supply (irrigated and rainfed) over three seasons. To complement field studies, a greenhouse study was conducted using plastic cylinders with soil inserted into PVC pipes, to determine the relationship between grain yield obtained under field conditions with different root traits measured under greenhouse conditions. Resistance to drought stress was positively associated with a deeper and vigorous root system, better shoot growth, and superior mobilization of photosynthates to pod and seed production. The drought resistant lines differed in their root characteristics, some of them with a vigorous and deeper root system while others with a moderate to shallow root system. Among the shoot traits measured, pod

  11. Quantitative X-ray - UV Line and Continuum Spectroscopy with Application to AGN: State-Specific Hydrogenic Recombination Cooling Coefficients for a Wide Range of Conditions

    Science.gov (United States)

    LaMothe, J.; Ferland, Gary J.

    2002-01-01

    Recombination cooling, in which a free electron emits light while being captured to an ion, is an important cooling process in photoionized clouds that are optically thick or have low metallicity. State specific rather than total recombination cooling rates are needed since the hydrogen atom tends to become optically thick in high-density regimes such as Active Galactic Nuclei. This paper builds upon previous work to derive the cooling rate over the full temperature range where the process can be a significant contributor in a photoionized plasma. We exploit the fact that the recombination and cooling rates are given by intrinsically similar formulae to express the cooling rate in terms of the closely related radiative recombination rate. We give an especially simple but accurate approximation that works for any high hydrogenic level and can be conveniently employed in large-scale numerical simulations.

  12. Germination and Early Growth of Brassica juncea in Copper Mine Tailings Amended with Technosol and Compost

    Science.gov (United States)

    González, Luís

    2014-01-01

    Mine tailings represent a serious threat to the environment and human health; thus their restoration has become a major concern. In this study, the interactions between Brassica juncea and different mine soil treatments were evaluated in order to understand their effect on germination and early growth. Three soil treatments containing 25% and 50% of technosol and 30% of compost were prepared. Germination and early growth were assessed in soil and pore water extracts from the treatments. Unlike the untreated mine soil, the three treatments allowed germination and growth, achieving levels comparable to those of seedlings from the same species developed in normal conditions. The seedlings grown in 50% of technosol and 30% of compost exhibited greater germination percentages, higher growth, and more efficient mechanisms against oxidative stress, ascribed to the organic matter and nutrients content of these treatments. Considering the unequivocal ability of B. juncea for phytoremediation, the results suggest that technosol and compost may be an auspicious solution to allow the germination and early growth of this species in mine tailings. PMID:25386602

  13. Phytoremediating a copper mine soil with Brassica juncea L., compost and biochar.

    Science.gov (United States)

    Rodríguez-Vila, Alfonso; Covelo, Emma F; Forján, Rubén; Asensio, Verónica

    2014-10-01

    The soils at a depleted copper mine in Touro (Galicia, Spain) are chemically degraded. In order to determine the effect of amendments and vegetation on the chemical characteristics of a mine soil and on the plant uptake of metals, a greenhouse experiment was carried out for 3 months. A settling pond soil was amended with different percentages of a compost and biochar mixture and vegetated with Brassica juncea L. The results showed that the untreated settling pond soil was polluted by Cu. Amendments and planting mustards decreased the pseudototal concentration of this metal, reduced the extreme soil acidity and increased the soil concentrations of C and TN. Both treatments also decreased the CaCl2-extractable Co, Cu and Ni concentrations. However, the amendments increased the pseudototal concentration of Zn in the soil, provided by the compost that was used. The results also showed that mustards extracted Ni efficiently from soils, suggesting that B. juncea L. is a good phytoextractor of Ni in mine soils.

  14. A Pin gene families encoding components of auxin efflux carriers in Brassica juncea

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Based on the sequence information of Arabidopsis PIN1, two cDNAs encoding PIN homologues fromBrassica juncea, Bjpin2 and Bjpin3, were isolated through cDNA library screening. Bjpin2 and Bjpin3encoded proteins containing 640 and 635 amino acid residues, respectively, which shared 97.5% identities witheach other and were highly homologous to Arabidopsis PIN1, PIN2 and other putative PIN proteins. BjPIN2and BjPIN3 had similar structures as AtPIN proteins. Northern blot analysis indicated that Bjpin2 wasexpressed in stem, leaf and floral tissues, while Bjpin3 was expressed predominantly in stem and hypocotyls.Two promoter fragments of pin genes, Bjpin-X and Bjpin-Z, were isolated by 'genome walking' techniqueusing primers at 5'-end of pin cDNA. Promoter-gus fusion studies revealed the GUS activities driven byBjpin-X were at internal side of xylem and petal; while those driven by Bjpin-Z were detected at leaf vein,epidermal cell and cortex of stem, vascular tissues and anther. Results of the pin genes with differentexpression patterns in B. juncea suggested the presence of a gene family.

  15. 28-Homobrassinolide Modulate Antenna Complexes and Carbon Skeleton of Brassica juncea L. under Temperature Stress

    Directory of Open Access Journals (Sweden)

    Harpreet Kaur

    2014-08-01

    Full Text Available The aim of present study was to explore the ameliorative impact of 28-homoBL on morpho-physiological attributes, photosynthetic pigments and sugars of Brassica juncea L. exposed to oxidative stress caused by extreme temperatures (4 and 44 °C. For this, experiments were carried out at the Plant Physiology Laboratory, Department of Botany, Punjabi University, Patiala. Effect of different degrees of temperature (4 and 44 °C taking 24 °C as control was studied. 28-homoBL (10-6, 10-9 and 10-12M primed and unprimed seeds of B. juncea L. in terms of antenna complexes and end products of photosynthesis that is total carbohydrates and total soluble sugars was investigated. All concentrations of 28-homoBL used in present study showed different effects on morphology and light quenching pigments. All concentrations of 28-homoBL showed promoting effect on growth and light quenching pigments. The carbon makeup ameliorated positively in stressed and non-stressed components of photosynthetic machinery and 10-9 M 28-homoBL showed best results. In conclusion 28-homoBL showed great potential in protecting the reaction centre of photosynthetic machinery from oxidative stress caused by extreme low and high temperatures but in very dose dependent manner and thus modulate the carbon skeleton of the plant.

  16. Growth responses of Brassica juncea to phosphorus application from different sources of fertilizer under salt stress

    Directory of Open Access Journals (Sweden)

    Badar-uz-Zaman

    2015-12-01

    Full Text Available Under salt-stress conditions, available phosphorus may not be in adequate amounts or in accessible forms for timely utilization by plants. This study was carried out to observe the growth and Na+ /K+ ratio in Brassica juncea having phosphorus supplied from commercial fertilizers under salt stress. Seeds of Brassica juncea (cv. Raya Anmol were germinated and the seedlings raised in standard nutrient solution under controlled conditions. Phosphorus was applied from three sources of fertilizers i.e. DAP, SSP and TSP @ 2 (as control and 10 mmol L-1. Salt stress was developed with NaCl (150 mmol L-1. Under salt stress and elevated P application as SSP and TSP, fresh mass was higher than with DAP source. Dry mass was higher with SSP source than with DAP and TSP. Na+ /K+ ratio was low using SSP. P-uptake by the plants was highest using TSP. For growth parameters and Na+ /K+ ratio, phosphate fertilizers contributed differentially under salt stress.

  17. Development of Cryopreservation Techniques for Gorgonian (Junceella juncea Oocytes through Vitrification.

    Directory of Open Access Journals (Sweden)

    Sujune Tsai

    Full Text Available Gorgonian corals are slowly declining due to human interaction and environmental impacts. Cryopreservation of gorgonian corals is an ex-situ method of conservation, ensuring future reproduction. The present study assessed the vitrification properties of cryoprotectant (CPT mixtures using the cryotop, cryoloop and open pulled straw (OPS cryopereservation methods prior to experimentation on gorgonian (Junceella juncea oocytes. Investigations of the equilibration and vitrification solutions' (ES and VS effect on oocytes throughout different incubation periods were conducted. The cryotop method was found to be the most successful in ensuring vitrification. The most favourable VS was composed of propylene glycol (PG, ethylene glycol (EG and methanol with concentrations of 3.5 M, 1.5 M and 2 M respectively. Experiments were performed using the cryotop method to cryopreserve Junceella juncea oocytes using VS2, the solution had the least impact on oocytes at 5°C rather than at 26°C. The success of the vitrification procedures was determined by adenosine triphosphate (ATP levels in cooled-thaw oocytes and the highest viability obtained from the present study was 76.6 ± 6.2%. This study provides information regarding gorgonian corals' tolerance and viability throughout vitrification to further advance the vitrification protocol on whip corals.

  18. Effect of cytokinins on shoot regeneration from cotyledon and leaf segment of stem mustard (Brassica juncea var. tsatsai)

    NARCIS (Netherlands)

    Guo, D.P.; Zhu, Z.J.; Hu, X.X.; Zheng Sijun, S.J.

    2005-01-01

    Cotyledon and leaf segments of stem mustard (Brassica juncea var. tsatsai) were cultured on Murashige and Skoog medium supplemented with various concentrations of different cytokinins [6-benzyladenine (BA), N-(2-chloro-4-pyridyl)-n-phenylurea (CPPU), 6-furfurylaminopurine (KT) and thidiazuron (TDZ)

  19. PEMANFAATAN LIMBAH KOTORAN SAPI DAN JERAMI KACANG TANAH SEBAGAI BOKASHI CAIR BAGI PERTUMBUHAN TANAMAN SAWI (Brassica juncea L.

    Directory of Open Access Journals (Sweden)

    Artiana Artiana

    2016-12-01

    Full Text Available Has conducted a study entitled " Cow Manure Waste Utilization and Straw Peanut For Liquid Bokashi For Plant Growth mustard (Brassica juncea L.". This study aimed to analyze the nutrient content in the liquid Bokashi is derived from cow dung and straw peanuts, and study the effect of dosing Bokashi different liquid to the growth of the mustard plant (Brassica juncea L.. Methods using a completely randomized design with one factor at a dose of 125 ml, 250 ml, 375 ml and 500 ml, and as control using Bokashi solid. This research was conducted with four replications. Bokashi liquid nutrient content of C of 0,1045%, 0,0461% of N, P and K amounted to 0.0172% at 0.2500%. The content of nutrient dense Bokashi C of 6,0874%, 2,0169% of N, P and K amounted to 0,0218% at 5,7802%. Bokashi dosing different liquid to the growth of mustard (Brassica juncea L. significant effect on the growth of mustard, but still lower than the solid Bokashi. Dose liquid Bokashi most optimal for growing mustard (Brassica juncea L. is at 375 ml.

  20. Effect of cytokinins on shoot regeneration from cotyledon and leaf segment of stem mustard (Brassica juncea var. tsatsai)

    NARCIS (Netherlands)

    Guo, D.P.; Zhu, Z.J.; Hu, X.X.; Zheng Sijun, S.J.

    2005-01-01

    Cotyledon and leaf segments of stem mustard (Brassica juncea var. tsatsai) were cultured on Murashige and Skoog medium supplemented with various concentrations of different cytokinins [6-benzyladenine (BA), N-(2-chloro-4-pyridyl)-n-phenylurea (CPPU), 6-furfurylaminopurine (KT) and thidiazuron (TDZ)

  1. Effect of Sunn Hemp (Crotalaria juncea L.) Cutting Date and Planting Density on Weed Suppression in Georgia, USA

    Science.gov (United States)

    A field study was conducted in 2008 and 2009 at the USDA, ARS, Plant Genetic Resources Conservation Unit in Griffin, GA to investigate weed suppression by sunn hemp (Crotalaria juncea L). The objectives were to: 1) evaluate the effects of apical meristem removal (AMR) at three dates [5, 6, and 7 wks...

  2. ‘Carolina Broadleaf’ mustard green (Brassica juncea L.) resistant to the bacterial leaf blight pathogen Pseudomonas cannabina pv. alisalensis

    Science.gov (United States)

    A leafy-green mustard (Brassica juncea L.) cultivar designated ‘Carolina Broadleaf’ has been released by the Agricultural Research Service of the U.S. Dept. of Agriculture in 2015. This released cultivar is a narrow-based population of leafy-green mustard derived from a U.S. plant introduction (PI)...

  3. Nitrogen-15 labeling of Crotalaria juncea green manure Marcação do adubo verde Crotalaria juncea com 15N

    Directory of Open Access Journals (Sweden)

    Edmilson José Ambrosano

    2003-02-01

    Full Text Available Most studies dealing with the utilization of 15N labeled plant material do not present details about the labeling technique. This is especially relevant for legume species since biological nitrogen fixation difficults plant enrichment. A technique was developed for labeling leguminous plant tissue with 15N to obtain labeled material for nitrogen dynamics studies. Sun hemp (Crotalaria juncea L. was grown on a Paleudalf, under field conditions. An amount of 58.32 g of urea with 70.57 ± 0.04 atom % 15N was sprayed three times on plants grown on eight 6-m²-plots. The labelled material presented 2.412 atom % 15N in a total dry matter equivalent to 9 Mg ha-1 This degree of enrichment enables the use of the green manure in pot or field experiments requiring 15N-labeled material.A grande maioria dos estudos com a utilização de material vegetal marcado com o isótopo 15N não apresentam detalhes tão importantes sobre como foram obtidos esses materiais. Em se tratando de marcação de leguminosas as dificuldades em se obter material marcado com 15N são ainda maiores pelo fato de serem plantas fixadoras de nitrogênio. Isso posto foi estabelecida uma técnica de marcação de leguminosas com nitrogênio (15N, com o objetivo de obter material vegetal marcado isotopicamente para estudos de dinâmica do nitrogênio. Cultivou-se a leguminosa crotalária júncea (Crotalaria juncea L., em Argissolo Vermelho Amarelo distrófico, em campo. Ao se aplicarem via foliar 58,32 gramas de uréia em oito canteiros experimentais, (uréia com 70,57 ± 0,04% de átomos de 15N parceladas em três vezes, obteve-se material vegetal marcado seco que continha 2,412 % em átomos de 15N em uma massa seca equivalente a 9 Mg ha-1. Essa marcação permite o uso dessa massa vegetal em estudos de dinâmica de nitrogênio.

  4. The role of phytochelatins and antioxidants in tolerance to Cd accumulation in Brassica juncea L.

    Science.gov (United States)

    Seth, Chandra Shekhar; Kumar Chaturvedi, Pranav; Misra, Virendra

    2008-09-01

    A hydroponics experiment using Indian mustard (Brassica juncea L.) was conducted to investigate the effect of different concentrations (10-160 microM) of cadmium (Cd) and a fixed concentration (500 microM) of ethylene diamine tetra acetic acid (EDTA) on Cd accumulation and its toxicity for 14 and 28 days (d). The results showed that Cd alone and Cd+EDTA increased total dry biomass production, photosynthetic pigments and total protein content of B. juncea up to 160 microM with respect to control for 14d (hormesis effect). Further, on treatment with Cd at 160 microM for 28d, dry biomass of root and shoot, total protein content and total chlorophyll decreased up to 73%, 58%, 67% and 53% respectively, while in the case of Cd+EDTA, the decrease in the above parameters was 38%, 50%, 57% and 46% with respect to their control. It was observed that the maximum Cd accumulation after 28d in the root and shoot was 1925 and 977 mg kg(-1) dry weight (dw), respectively, while in the case of Cd+EDTA it was 1013 and 2316 mg kg(-1)dw, respectively. Levels of phytochelatins (PCs), glutathione reductase (GR; EC 1.6.4.2), non-protein thiols (NP-SH) and glutathione (GSH) were monitored as plants primary and secondary metal detoxifying responses. Glutathione reductase showed three-fold increased activity for Cd and 2.2-fold for Cd+EDTA at 160 microM after 14d followed by decreased activity after 28d with respect to control. Maximum synthesis of PCs was found at 10 microM of Cd exposure followed by a gradual decline after 28d. This may be correlated with reduced level of GSH, probably due to reduced GR activity, resulting in enhanced oxidative stress as also proved by phenotypic changes in plants such as browning of roots and yellowing of leaves. Thus, the capacity of B. juncea to accumulate and tolerate high concentrations of Cd, through enhanced level of PCs, GSH, NP-SH and GR suggests its applicability for phytoremediation.

  5. In Vivo Rescue of a Silent tax-Deficient Bovine Leukemia Virus from a Tumor-Derived Ovine B-Cell Line by Recombination with a Retrovirally Transduced Wild-Type tax Gene

    Science.gov (United States)

    Van Den Broeke, Anne; Bagnis, Claude; Ciesiolka, Malgorzata; Cleuter, Yvette; Gelderblom, Hans; Kerkhofs, Pierre; Griebel, Philip; Mannoni, Patrice; Burny, Arsene

    1999-01-01

    The lack of bovine leukemia virus (BLV) expression is a consistent finding in freshly isolated ovine tumor cells and in the B-cell lines derived from these tumors. In order to gain further insight into the mechanisms of BLV silencing in these tumors, we have used the YR2 B-cell line, which was derived from the leukemic cells of a BLV-infected sheep. This cell line contains a single, monoclonally integrated, silent provirus, which cannot be reactivated either by stimulation in vitro or by in vivo injection of the tumor cells or cloned proviral DNA in sheep. Sequence analysis of the tax gene from the YR2 cell line identified two G-to-A transitions (G7924 to A7924 and G8149 to A8149) that result in E-to-K amino acid changes at positions 228 and 303 in the Tax protein. Following retroviral vector-mediated transfer of a wild-type tax gene into YR2 cells, we showed that BLV mRNA, viral proteins, and virions were produced, demonstrating that the cellular factors required for virus expression were present in the original YR2 cell line. Injection of this transduced YR2 cell line in sheep led to the rescue of replication-competent BLV proviruses. The integrated competent proviruses exhibited unique chimeric tax genes, which arose from homologous recombination between the transduced wild-type tax and the YR2-derived tax sequences. Furthermore, in one of these functional recombinant proviruses, only the A8149-to-G8149 reversion was present, providing clear evidence that the defect underlying the silent phenotype in YR2 cells results from a single C-terminal E303-to-K303 amino acid substitution in the BLV Tax protein. Our observations suggest that a single strategically located mutation in tax provides a mechanism for BLV inactivation in B-cell tumors. PMID:9882306

  6. Recombination instability

    DEFF Research Database (Denmark)

    D'Angelo, N.

    1967-01-01

    A recombination instability is considered which may arise in a plasma if the temperature dependence of the volume recombination coefficient, alpha, is sufficiently strong. Two cases are analyzed: (a) a steady-state plasma produced in a neutral gas by X-rays or high energy electrons; and (b) an af...

  7. Biological activities of recombinant equine luteinizing hormone/chorionic gonadotropin (eLH/CG) expressed in Sf9 and Mimic insect cell lines.

    Science.gov (United States)

    Legardinier, Sébastien; Duonor-Cérutti, Martine; Devauchelle, Gérard; Combarnous, Yves; Cahoreau, Claire

    2005-02-01

    Equine luteinizing hormone (eLH) and chorionic gonadotropin (eCG) are composed of identical alpha and beta polypeptide chains, but eCG subunits are much more heavily glycosylated and sialylated. Consequently, eCG exhibits a much longer half-life than eLH in blood. Recombinant eLH/CG, expressed in Sf9 and Mimic insect cells, were compared with one another and to the natural hormones eCG and eLH. Mimic cells are stably-transformed Sf9 cells, expressing five mammalian genes encoding glycosyltransferases involved in the synthesis of complex N-carbohydrate chains. Recombinant eLH/CG expressed in Mimic cells exhibited a higher apparent molecular weight (MW) than that expressed in Sf9 cells, suggesting that its N-glycosylation was, as expected, more complete. Nevertheless, the two recombinant eLH/CG exhibited lower MW than natural eCG from pregnant mare plasma. The two eLH/CG produced in Sf9 and Mimic cells were found to be active in in vitro LH and FSH bioassays, with potencies similar to those of eCG. By contrast, they exhibited no significant in vivo bioactivity, neither in the specific follicle-stimulating hormone (FSH) assay nor in the specific eCG assay. Although recombinant eLH/CG produced in Mimic cells bears more elaborate carbohydrate chains than recombinant eLH/CG from Sf9 cells, it exhibits no significant in vivo bioactivity, probably because of insufficient terminal sialylation of its carbohydrate chains, leading to its rapid removal from blood.

  8. QTL analysis of novel genomic regions associated with yield and yield related traits in new plant type based recombinant inbred lines of rice (Oryza sativa L.

    Directory of Open Access Journals (Sweden)

    Marathi Balram

    2012-08-01

    Full Text Available Abstract Background Rice is staple food for more than half of the world’s population including two billion Asians, who obtain 60-70% of their energy intake from rice and its derivatives. To meet the growing demand from human population, rice varieties with higher yield potential and greater yield stability need to be developed. The favourable alleles for yield and yield contributing traits are distributed among two subspecies i.e., indica and japonica of cultivated rice (Oryza sativa L.. Identification of novel favourable alleles in indica/japonica will pave way to marker-assisted mobilization of these alleles in to a genetic background to break genetic barriers to yield. Results A new plant type (NPT based mapping population of 310 recombinant inbred lines (RILs was used to map novel genomic regions and QTL hotspots influencing yield and eleven yield component traits. We identified major quantitative trait loci (QTLs for days to 50% flowering (R2 = 25%, LOD = 14.3, panicles per plant (R2 = 19%, LOD = 9.74, flag leaf length (R2 = 22%, LOD = 3.05, flag leaf width (R2 = 53%, LOD = 46.5, spikelets per panicle (R2 = 16%, LOD = 13.8, filled grains per panicle (R2 = 22%, LOD = 15.3, percent spikelet sterility (R2 = 18%, LOD = 14.24, thousand grain weight (R2 = 25%, LOD = 12.9 and spikelet setting density (R2 = 23%, LOD = 15 expressing over two or more locations by using composite interval mapping. The phenotypic variation (R2 ranged from 8 to 53% for eleven QTLs expressing across all three locations. 19 novel QTLs were contributed by the NPT parent, Pusa1266. 15 QTL hotpots on eight chromosomes were identified for the correlated traits. Six epistatic QTLs effecting five traits at two locations were identified. A marker interval (RM3276-RM5709 on chromosome 4 harboring major QTLs for four traits was identified. Conclusions The present study reveals that favourable alleles for

  9. A novel technique to collect root exudates from mustard (Brassica juncea).

    Science.gov (United States)

    Wadhwa, Kunal; Narula, Neeru

    2012-10-01

    A very simple, novel, cost effective, easy to use technique has been developed for the collection of root exudates from small seeded plants, under laboratory conditions. 200-1000 μl micro tips (Tarsons), kept in 100 ml glass beakers, were used as holders for the small seeds of Indian mustard (Brassica juncea) and the exudates were trapped in liquid culture medium. The exudates, so obtained, were authenticated and analyzed for organic compounds such as sugars, amino acids and organic acids, as well as chemotactic response towards rhizobacteria. Method was found to be suitable and easy to handle for small seeds. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Efek Paparan Musik dan Noise pada Karakteristik Morfologi dan Produktivitas Tanaman Sawi Hijau (Brassica Juncea

    Directory of Open Access Journals (Sweden)

    Joko Prasetyo

    2014-04-01

    Full Text Available The objective of present study was to investigate the effect of various sounds on the green mustard’s (Brassica Juncea morphology characteristic and productivity. The plant has been subjected to three various sound, namely classical music (rhythmic violin music, machine and traffic noise, and mixed sound (classical music and traffic noise with 70-75 dB sound pressure level, from germination to harvest for three hours (7-10 am. each day. Six parameters, i.e. germination, plant height, leaf width, leaf lenght, total plant lenght, and fresh weight, related with growth and productivity of plant were been monitored on regular basis.The results showed classical music improves germination up to 15% for 36 hours, plant height 13,5%, leaf width 14,8%, leaf length 14,2%, and wet weight 57,1%. In general, exposure to classical music gives the best results on the morphological characteristics and productivity of green mustard.

  11. Cadmium stress responses in Brassica juncea: hints from proteomics and metabolomics.

    Science.gov (United States)

    D'Alessandro, Angelo; Taamalli, Manel; Gevi, Federica; Timperio, Anna Maria; Zolla, Lello; Ghnaya, Tahar

    2013-11-01

    Among heavy metal stressors, cadmium (Cd) pollution is one leading threat to the environment. In this view, research efforts have been increasingly put forward to promote the individuation of phytoextractor plants that are capable of accumulating and withstanding the toxic metals, including Cd, in the aerial parts. We hereby adopted the hyperaccumulator B. juncea (Indian mustard) as a model to investigate plant responses to Cd stress at low (25 μM) and high (100 μM) doses. Analytical strategies included mass-spectrometry-based determination of Cd and the assessment of its effect on the leaf proteome and metabolome. Results were thus integrated with routine physiological data. Taken together, physiology results highlighted the deregulation of photosynthesis efficiency, ATP synthesis, reduced transpiration, and the impairment of light-independent carbon fixation reactions. These results were supported at the proteomics level by the observed Cd-dependent alteration of photosystem components and the alteration of metabolic enzymes, including ATP synthase subunits, carbonic anhydrase, and enzymes involved in antioxidant responses (especially glutathione and phytochelatin homeostasis) and the Calvin cycle. Metabolomics results confirmed the alterations of energy-generating metabolic pathways, sulfur-compound metabolism (GSH and PCs), and Calvin cycle. Besides, metabolomics results highlighted the up-regulation of phosphoglycolate, a byproduct of the photorespiration metabolism. This was suggestive of the likely increased photorespiration rate as a means to cope with Cd-induced unbalance in stomatal conductance and deregulation of CO2 homeostasis, which would, in turn, promote CO2 depletion and O2 (and thus oxidative stress) accumulation under prolonged photosynthesis in the leaves from plants exposed to high doses of CdCl2. Overall, it emerges that Cd-stressed B. juncea might rely on photorespiration, an adaptation that would prevent the over-reduction of the

  12. Sulfur utilization by rice and Crotalaria juncea from sulfate - 34S applied to the soil

    Directory of Open Access Journals (Sweden)

    Trivelin Paulo Cesar Ocheuze

    2002-01-01

    Full Text Available In tropical soils with intensive agriculture an increasing sulfur deficiency has been verified in several crops. The low available S in these soils is caused by the continuous use of concentrated NPK fertilizers. The objective of this work was to evaluate the utilization by rice (Oriza sativa L. and crotalaria juncea (Crotalaria juncea L. of sulfur applied to the soil, under greenhouse conditions. Pots with 3 kg of an Argisol (Paleudalf were used to test the isotopic technique with the stable isotope 34S, adding a solution of sodium sulfate labeled with 34S (14.30 ± 0.05 atom % of 34S to the soil (70 mg SO4-S per kg-1 of soil 18 days after sowing both species. The shoots of the crotalaria and rice were harvested, respectively on the 72nd and 122nd days after S fertilization. The concentration and the amount of sulfur in the crotalaria were higher than in rice, due to the higher legume requirement for this nutrient. The sulfur requirement and the short time interval between fertilization and harvest of the crotalaria resulted in a small amount of native SO4-S mineralized in the soil and a small quantity of 34SO4 immobilized by soil microorganisms. Thus, the percentage of sulfur in the crotalaria derived from the fertilizer (Sdff was higher than in the rice (%Sdff crotalaria = 91.3 ± 3.5%; %Sdff rice = 66.3 ± 0.8%. The expressive values of %Sdff indicate a low rate of mineralization of SO4-S probably as a consequence of the low available sulfur content in the soil.

  13. Recombination monitor

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, S. Y. [Brookhaven National Lab. (BNL), Upton, NY (United States); Blaskiewicz, M. [Brookhaven National Lab. (BNL), Upton, NY (United States)

    2017-02-03

    This is a brief report on LEReC recombination monitor design considerations. The recombination produced Au78+ ion rate is reviewed. Based on this two designs are discussed. One is to use the large dispersion lattice. It is shown that even with the large separation of the Au78+ beam from the Au79+ beam, the continued monitoring of the recombination is not possible. Accumulation of Au78+ ions is needed, plus collimation of the Au79+ beam. In another design, it is shown that the recombination monitor can be built based on the proposed scheme with the nominal lattice. From machine operation point of view, this design is preferable. Finally, possible studies and the alternative strategies with the basic goal of the monitor are discussed.

  14. Transgenic Indian mustard (Brassica juncea) plants expressing an Arabidopsis phytochelatin synthase (AtPCS1) exhibit enhanced As and Cd tolerance.

    Science.gov (United States)

    Gasic, Ksenija; Korban, Schuyler S

    2007-07-01

    Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides that play important roles in detoxification of heavy metal and metalloids in plants and other living organisms. The overall goal of this study is to develop transgenic plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian mustard, and transgenic pcs lines have been compared with wild-type plants for tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild type plants, transgenic plants exhibit significantly higher tolerance to Cd and As. Shoots of Cd-treated pcs plants have significantly higher concentrations of PCs and thiols than those of wild-type plants. Shoots of wild-type plants accumulated significantly more Cd than those of transgenic plants, while accumulation of As in transgenic plants was similar to that in wild type plants. Although phytochelatin synthase improves the ability of Indian mustard to tolerate higher levels of the heavy metal Cd and the metalloid As, it does not increase the accumulation potential of these metals in the above ground tissues of Indian mustard plants.

  15. Comparison of the karyotypes ofPsathyrostachys juncea andP. huashanica (Poaceae) studied by banding techniques

    DEFF Research Database (Denmark)

    Linde-Laursen, Ib; Bothmer, R. von

    1986-01-01

    . The patterns of both taxa are polymorphic, supporting that both taxa are outbreeders. The karyotypic characters suggest that P. juncea is more closely related to P. fragilis than either is to P. huashanica. N-banding stains weakly. Silver nitrate staining demonstrates that nucleolus organizers of both species...... have different nucleolus forming capacities. The presence of micronucleoli suggests that both species have an extra unidentified chromosome with nucleolus forming capacity....

  16. Acute pulmonary emphysema cum pulmonary edema apparently associated with feeding of Brassica juncea in a dairy buffalo

    OpenAIRE

    Muhammad, Ghulam; Saqib, Muhammad; Naureen, Abeera

    2010-01-01

    This preliminary report describes the occurrence of acute pulmonary emphysema cum pulmonary edema ensuing in extensive subcutaneous emphysematous swellings in a dairy buffalo (Bubalus bubalis) apparently associated with a sudden shift from berseem (Trifolium alexendrinum) to Brassica juncea fodder. Tachypnea, expiratory dyspnea, open-mouth breathing, loud expiratory grunt with abdominal lift, and crackles in ventral aspects of the lungs with normal rectal temperature characterized the conditi...

  17. Castasterone confers copper stress tolerance by regulating antioxidant enzyme responses, antioxidants, and amino acid balance in B. juncea seedlings.

    Science.gov (United States)

    Yadav, Poonam; Kaur, Ravdeep; Kanwar, Mukesh Kumar; Sharma, Anket; Verma, Vinod; Sirhindi, Geetika; Bhardwaj, Renu

    2017-09-20

    The aim of the present study was to explore the effect of exogenous application of castasterone (CS) on physiologic and biochemical responses in Brassica juncea seedlings under copper (Cu) stress. Seeds were pre-soaked in different concentrations of CS and grown for 7 days under various levels of Cu. The exposure of B. juncea to higher levels of Cu led to decrease of morphologic parameters, with partial recovery of length and fresh weight in the CS pre-treated seedlings. Metal content was high in both roots and shoots under Cu exposure while the CS pre-treatment reduced the metal uptake. Accumulation of hydrogen peroxide (H2O2) and superoxide anion radical (O2(-)) were chosen as stress biomarker and higher levels of H2O2 (88.89%) and O2(-) (62.11%) showed the oxidative stress in metal treated B. juncea seedlings, however, CS pre-treatment reduced ROS accumulation in Cu-exposed seedlings. The Cu exposures lead to enhance the plant's enzymatic and non-enzymatic antioxidant system. It was observed that enzymatic activities of ascorbate peroxidase (APOX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), glutathione perxoidase (GPOX) and gultrathione-s-transferase increased while activity of monodehydroascorbate reductase (MDHAR) decreased under Cu stress. The pre-treatment with CS positively affected the activities of enzymes. RT-PCR analysis showed that mRNA transcript levels were correlated with total enzymatic activity of DHAR, GR, GST and GSH. Increase in the gene expression of DHAR (1.85 folds), GR (3.24 folds), GST-1 (2.00 folds) and GSH-S (3.18 folds) was noticed with CS pre-treatment. Overall, the present study shows that Cu exposure induced severe oxidative stress in B. juncea plants and exogenous application of CS improved antioxidative defense system by modulating the ascorbate-glutathione cycle and amino acid metabolism. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. 24-epibrassinolide mitigates the adverse effects of manganese induced toxicity through improved antioxidant system and photosynthetic attributes in Brassica juncea.

    Science.gov (United States)

    Fariduddin, Qazi; Ahmed, Mumtaz; Mir, Bilal A; Yusuf, Mohammad; Khan, Tanveer A

    2015-08-01

    The objective of this study was to establish relationship between manganese-induced toxicity and antioxidant system response in Brassica juncea plants and also to investigate whether brassinosteroids activate antioxidant system to confer tolerance to the plants affected with manganese induced oxidative stress. Brassica juncea plants were administered with 3, 6, or 9 mM manganese at 10-day stage for 3 days. At 31-day stage, the seedlings were sprayed with deionized water (control) or 10(-8) M of 24-epibrassinolide, and plants were harvested at 45-day stage to assess growth, leaf gas-exchange traits, and biochemical parameters. The manganese treatments diminished growth along with photosynthetic attributes and carbonic anhydrase activity in the concentration-dependent manner, whereas it enhanced lipid peroxidation, electrolyte leakage, accumulation of H2O2 as well as proline, and various antioxidant enzymes in the leaves of Brassica juncea which were more pronounced at higher concentrations of manganese. However, the follow-up application of 24-epibrassinolide to the manganese stressed plants improved growth, water relations, and photosynthesis and further enhanced the various antioxidant enzymes viz. catalase, peroxidase, and superoxide dismutase and content of proline. The elevated level of antioxidant enzymes as well as proline could have conferred tolerance to the manganese-stressed plants resulting in improved growth and photosynthetic attributes.

  19. Inoculation of plant growth promoting bacterium Achromobacter xylosoxidans strain Ax10 for the improvement of copper phytoextraction by Brassica juncea.

    Science.gov (United States)

    Ma, Ying; Rajkumar, Mani; Freitas, Helena

    2009-02-01

    In this study, a copper-resistant plant growth promoting bacterial (PGPB) strain Ax10 was isolated from a Cu mine soil to assess its plant growth promotion and copper uptake in Brassica juncea. The strain Ax10 tolerated concentrations up to 600 mg CuL(-1) on a Luria-Bertani (LB) agar medium and utilized 1-aminocyclopropane-1-carboxylic acid (ACC) as a sole N source in DF salts minimal medium. The strain Ax10 was characterized as Achromobacter xylosoxidans based on its 16S rDNA sequence homology (99%). The bacterium A. xylosoxidans Ax10 has also exhibited the capability of producing indole acetic acid (IAA) (6.4 microg mL(-1)), and solubilizing inorganic phosphate (89.6 microg mL(-1)) in specific culture media. In pot experiments, inoculation of A. xylosoxidans Ax10 significantly increased the root length, shoot length, fresh weight and dry weight of B. juncea plants compared to the control. This effect can be attributed to the utilization of ACC, production of IAA and solubilization of phosphate. Furthermore, A. xylosoxidans Ax10 inoculation significantly improved Cu uptake by B. juncea. Owing to its wide action spectrum, the Cu-resistant A. xylosoxidans Ax10 could serve as an effective metal sequestering and growth promoting bioinoculant for plants in Cu-stressed soil. The present study has provided a new insight into the phytoremediation of Cu-contaminated soil.

  20. Understanding the biology and ecology of vulnerable plant species: case study with tetratheca juncea occurring over coal leases

    Energy Technology Data Exchange (ETDEWEB)

    David Mulligan; Sean Bellairs; F.V. Bartier; C.L. Gross; D. Bowen

    2001-06-01

    Tetratheca juncea Smith (Tremandraceae) is a vulnerable species listed under the NSW Threatened Species Conservation Act (Schedule 2, TSC Act 1995), and in the Commonwealth Environment Protection and Biodiversity Conservation Act 1999.Researchers at the Universities of Queensland, New England and Newcastle established A collaborative research program investigated the reproductive and establishment biology of T juncea. Breeding systems, seed biology and mycorrhizal associations were investigated to determine factors limiting the reproductive output of the species. Native bees necessary for pollination were not detected in 100 hours of observation. The three key ramifications from this study of T. juncea's ecology is that: a pollinator is required for high seed yields; fire is required for germination; and a mycorrhizal partner is required for plant longevity. These findings indicate that translocations of the species cannot be recommended as there is a lack of knowledge about many factors that are critical for the persistence of the species. A fire management plan will need to cater for all obligate ecological requirements. The results of this study have been used to develop a flowchart on the biological procedures that need to be considered when a threatened flora species is found on a site. The results from this study are also considered to be a relevant guide for managing populations of other species of Tetratheca, many of which are also rare or threatened.

  1. In vitro anti-biofilm and anti-bacterial activity of Junceella juncea for its biomedical application

    Institute of Scientific and Technical Information of China (English)

    P Kumar; S Senthamil Selvi; M Govindaraju

    2012-01-01

    Objective: To investigate the anti-biofilm and anti-bacterial activity of Junceella juncea (J. juncea) against biofilm forming pathogenic strains. Methods: Gorgonians were extracted with methanol and analysed with fourier transform infrared spectroscopy. Biofilm forming pathogens were identified by Congo red agar supplemented with sucrose. A quantitative spectrophotometric method was used to monitor in vitro biofilm reduction by microtitre plate assay. Anti-bacterial activity of methanolic gorgonian extract (MGE) was carried out by disc diffusion method followed by calculating the percentage of increase with crude methanol (CM). Results: The presence of active functional group was exemplified by FT-IR spectroscopy. Dry, black, crystalline colonies confirm the production of extracellular polymeric substances responsible for biofilm formation in Congo red agar. MGE exhibited potential anti-biofilm activity against all tested bacterial strains. The anti-bacterial activity of methanolic extract was comparably higher in Salmonella typhii followed by Escherichia coli, Vibrio cholerae and Shigella flexneri. The overall percentage of increase was higher by 50.2%to CM. Conclusions:To conclude, anti-biofilm and anti-bacterial efficacy of J. juncea is impressive over biofilm producing pathogens and are good source for novel anti-bacterial compounds.

  2. Serum-free culture of an embryonic cell line from Bombyx mori and reinforcement of susceptibility of a recombinant BmNPV by cooling.

    Science.gov (United States)

    Imanishi, Shigeo; Kobayashi, Jun; Sekine, Toshiaki

    2012-03-01

    We established the first continuous cell line that uses a serum-free culture from the embryo of Bombyx mori (Lepidoptera: Bombycidae), designated as NIAS-Bm-Ke17. This cell line was serially subcultured in the SH-Ke-117 medium. The cells adhere weakly to the culture flask, and most cells have an oval shape. The cell line was subcultured 154 times, and the population doubling time is 83.67±5.22 h. Random amplification of polymorphic DNA-polymerase chain reaction with a tenmar single primer for discrimination of insect cell lines recognized the NIAS-Bm-Ke1 cell line as B. mori. This cell line does not support the growth of the B. mori nuclear polyhedrosis virus (BmNPV) in the absence of the heat-inactivated hemolymph of B. mori. However, the heat-inactivated hemolymph in 1% volume of the medium supported a high level of susceptibility to BmNPV. In addition, the cooling treatment of the cells at 2.5°C also enhanced the susceptibility. We report a new serum-free culture system of the B. mori cell line for the baculovirus expression vector system.

  3. Variabilidade espacial de atributos de solo e de Crotalaria juncea L. em área severamente erodida

    Directory of Open Access Journals (Sweden)

    A. A. C. Salviano

    1998-03-01

    Full Text Available O trabalho foi desenvolvido numa área de produção comercial de cana-de-açúcar, situada no município de Piracicaba (SP, numa associação de Podzólico Vermelho-Amarelo + solo litólico, no período de novembro de 1994 a março de 1995. Essa área vem sendo explorada com cana-de-açúcar há, aproximadamente, 30 anos e apresentava diversos sulcos de erosão. O objetivo deste trabalho foi caracterizar a variabilidade espacial de atributos de crotalária juncea (Crotalaria juncea L. e de solo em uma área sob condições de erosão severa. A área foi arada e gradeada com incorporação de 4 t ha-1 de calcário, antes da semeadura a lanço de 30 kg ha-1 de semente de crotalária juncea. Uma parcela de 50 x 70 m foi amostrada de acordo com uma malha de 5 por 5 m, totalizando 140 pontos. Foram avaliados atributos químicos do solo superficial (0,00-0,20 m e subsuperficial (0,20-0,40 m, sua granulometria e a espessura de solo remanescente (ES - definida como a camada do solo acima do horizonte C, além da produtividade de matéria seca (MS e altura da crotalária juncea (ALTPL. Os valores de atributos maiores ou menores que quatro desvios-padrões da média foram descartados. A colheita da parte aérea da crotalária foi realizada no início de sua floração, em miniparcelas de 2 x 2,5 m, e calculada a matéria seca. A dependência espacial dos atributos estudados foi avaliada por semivariogramas escalonados. Esses apresentaram dependência espacial, com exceção do P (0,00-0,20 m e K nas duas camadas. Os atributos puderam ser agrupados em três categorias homogêneas quanto ao alcance do semivariograma: atributos químicos do solo (12 a 32 m < componentes de planta (25 a 32 m < frações granulométricas (32 a 42 m. Os atributos que melhor explicaram a produtividade da crotalária juncea foram H + Al, valor T e saturação por bases.

  4. Transcriptional activation and localization of expression of Brassica juncea putative metal transport protein BjMTP1

    Directory of Open Access Journals (Sweden)

    Salt David E

    2007-06-01

    Full Text Available Abstract Background Metal hyperaccumulators, including various Thlaspi species, constitutively express the putative metal transporter MTP1 to high levels in shoots. Here we present data on the transcriptional regulation and localization of expression of the homologous gene BjMTP1 in Brassica juncea. Though B. juncea lacks the ability to hyperaccumulate metals, its relatively high biomass, rapid growth and relatedness to true metal hyperaccumulating plants makes it a promising starting point for the development of plants for phytoremediation. Our goal in this study is to determine the transcriptional regulation of MTP1 in order to start to better understanding the physiological role of MTP1 in B. juncea. Results Steady-state mRNA levels of BjMTP1 were found to be enhanced 8.8, 5.9, and 1.6-fold in five-day-old B. juncea seedlings after exposure to Ni2+, Cd2+ or Zn2+, respectively. This was also reflected in enhanced GUS activity in B. juncea seedlings transformed with BjMTP1 promoter::GUSPlus after exposure to these metals over a similar range of toxicities from mild to severe. However, no increase in GUS activity was observed after exposure of seedlings to cold or heat stress, NaCl or hydrogen peroxide. GUS expression in Ni2+ treated seedlings was localized in roots, particularly in the root-shoot transition zone. In four- week- old transgenic plants BjMTP1 promoter activity also primarily increased in roots in response to Ni2+ or Cd2+ in plants transformed with either GUS or mRFP1 as reporter genes, and expression was localized to the secondary xylem parenchyma. In leaves, BjMTP1 promoter activity in response to Ni2+ or Cd2+ spiked after 24 h then decreased. In shoots GUS expression was prominently present in the vasculature of leaves, and floral parts. Conclusion Our studies establish that a 983 bp DNA fragment upstream of the BjMTP1 translational start site is sufficient for the specific activation by Ni2+ and Cd2+ of BjMTP1 expression

  5. Transcriptional activation and localization of expression of Brassica juncea putative metal transport protein BjMTP1

    Science.gov (United States)

    Muthukumar, Balasubramaniam; Yakubov, Bakhtiyor; Salt, David E

    2007-01-01

    Background Metal hyperaccumulators, including various Thlaspi species, constitutively express the putative metal transporter MTP1 to high levels in shoots. Here we present data on the transcriptional regulation and localization of expression of the homologous gene BjMTP1 in Brassica juncea. Though B. juncea lacks the ability to hyperaccumulate metals, its relatively high biomass, rapid growth and relatedness to true metal hyperaccumulating plants makes it a promising starting point for the development of plants for phytoremediation. Our goal in this study is to determine the transcriptional regulation of MTP1 in order to start to better understanding the physiological role of MTP1 in B. juncea. Results Steady-state mRNA levels of BjMTP1 were found to be enhanced 8.8, 5.9, and 1.6-fold in five-day-old B. juncea seedlings after exposure to Ni2+, Cd2+ or Zn2+, respectively. This was also reflected in enhanced GUS activity in B. juncea seedlings transformed with BjMTP1 promoter::GUSPlus after exposure to these metals over a similar range of toxicities from mild to severe. However, no increase in GUS activity was observed after exposure of seedlings to cold or heat stress, NaCl or hydrogen peroxide. GUS expression in Ni2+ treated seedlings was localized in roots, particularly in the root-shoot transition zone. In four- week- old transgenic plants BjMTP1 promoter activity also primarily increased in roots in response to Ni2+ or Cd2+ in plants transformed with either GUS or mRFP1 as reporter genes, and expression was localized to the secondary xylem parenchyma. In leaves, BjMTP1 promoter activity in response to Ni2+ or Cd2+ spiked after 24 h then decreased. In shoots GUS expression was prominently present in the vasculature of leaves, and floral parts. Conclusion Our studies establish that a 983 bp DNA fragment upstream of the BjMTP1 translational start site is sufficient for the specific activation by Ni2+ and Cd2+ of BjMTP1 expression primarily in roots. Activation of

  6. PEMANFAATAN PESTISIDA NABATI PADA PENGENDALIAN HAMA PLUTELLA XYLOSTELLA TANAMAN SAWI (Brassica juncea L. MENUJU PERTANIAN RAMAH LINGKUNGAN

    Directory of Open Access Journals (Sweden)

    suhartini suhartini

    2017-05-01

    ABSTRACT The use of chemical pesticides has many negative impacts on the environment, it is necessary for the use of biological pesticide towards sustainable agriculture or environmentally friendly agriculture. In the village were encountered many types of leaves that can be used as a biological pesticide, and therefore this study aims to determine the effectiveness of pesticide plant extracts of the leaves of some plants covering the leaves of tobacco, elephantopus, yellow wood and green betel on mortality of Plutella xylostella pests in plants mustard greens (Brassica juncea L, heavy wet mustard and mustard leaf damage    This study uses a completely randomized design with the treatment of various crops as a pesticide vegetable. The treatments used 6 kinds (degree ie negative control (P0, the leaves of tobacco (P1, the leaves of elephantopus (P2, the leaves of yellow wood (P3, the leaves of greens betel (P4 and chemical pesticides as a positive control (P5 with each grade of 10 %. Parameters measured were mortality pests, heavy wet mustard greens and mustard greens leaf damage rate. The analysis was performed by one-way ANOVA.    The results showed that the extract from the leaves of tobacco (Nicotiana tabacum, the leaves of elephantopus, the leaves of yellow wood (Arcangelisia flava L. and the leaves of green betel (Piper betle Linn. With a 10 percent concentration of the extract on a variety of  leaves are not yet significant effect on mortality pests Plutella xylostella, heavy wet of mustard greens (brassica juncea l. and severity of leaf mustard greens (brassica juncea l.. The leaf extract the most influence on mortality Plutella xylostella sequential is an extract of the leaves tobacco, the leaves of greens betel, the leaves of yellow wood and the leaves of elephantopus. While the effect on weight of wet mustard greens (Brassica juncea l. in order are as follows: the leaves of yellow wood, the leaves of green betel, the leaves of elephantopus, and the

  7. Recombinant fowl poxvirus vUTAL3CP1 multiplicationin in a mammalian cell line%鸡痘病毒在哺乳细胞内复制的研究

    Institute of Scientific and Technical Information of China (English)

    金明兰; 金宁一; 鲁会军; 马鸣潇; 郑敏

    2013-01-01

    Fowl poxviruses have many advantages and are being increasingly employed as recombinant vaccine vectors. One attractive feature is that while inserted transgenes are expressed in im-munologically favourable ways, fowl poxvirus infections of mammalian cells are believed to be a-bortive. The experimental evidence supporting this belief is, however, based on a limited number of mammalian cell-types and a few fowl poxvirus species. We evaluated two avian and 13 mammalian cell lines permissively to recombinant fowl poxvirus strains. Both avian cell lines were,as expected, permissive for recombinant fowl poxvirus strains. However, by multiplication assays, we found to our surprise that Syrian baby hamster kidney (BHK-21),pig kidney (PK),bovine testic-ular (BT), bovine embryo kidney (MDBK ) and bovine embryo trachea (EBTR) cells were equally permissive to recombinant strains. Results from electron microscopy of infected BHK-21,PK, BTC cells revealed viral morphogenesis proceeding to various forms of infectious viruses. These results were supported by the demonstration of fowl poxvirus specific late gene expression and fowl poxvirus specific DNA restriction pattern in BHK-21 infected cells. The results showed that the live attenuated virus adapted to the PK,BTC were safe as vaccine.%将重组鸡痘病毒在1种禽类细胞和13种哺乳动物及人的细胞内培养.通过电镜观察可见重组鸡痘病毒在CEE、BHK、PK、BTC、Vero细胞内的复制,同时用PCR方法扩增出目的基因,Western-blotting检测到目的蛋白,验证是鸡痘病毒.试验表明鸡痘病毒在禽类、个别哺乳动物细胞中可以复制,但尚未发现在人的细胞内复制;在CEF中可以稳定传30代以上;在BHK内第1代可以观察到细胞病变(CPE);在PK、BTC、Vero内第2代可以观察到CPE,证明鸡痘病毒在个别哺乳动物细胞发生复制,但不能稳定遗传.

  8. Cadmium tolerance and its phytoremediation by two oil yielding plants Ricinus communis (L.) and Brassica juncea (L.) from the contaminated soil.

    Science.gov (United States)

    Bauddh, Kuldeep; Singh, Rana P

    2012-09-01

    The effect of increasing level of cadmium in soil was investigated on biomass production, antioxidants, Cd bioaccumulation and translocation in Ricinus communis vis-à-vis a commonly studied oil crop Brassica juncea. The plants were exposed to 25, 50, 75, 100, and 150 mg Cd/Kg soil for up to 60 days. It was found that R. communis produced higher biomass at all the contamination levels than that of B. juncea. Proline and malondialdehyde in the leaves increased with increase in Cd level in both the species, whereas soluble protein decreased. The bioaccumulation of Cd was higher in B. juncea on the basis of the per unit biomass, total metal accumulation per plant was higher in R. communis. The translocation of Cdfrom roots to shoot was also higher in B. juncea at all Cd concentrations. R. communis appeared more tolerant and capable to clean Cd contaminated soil for longer period in one sowing than B. juncea and the former can grow in wasteland soil also in which later cannot be cultivated.

  9. Abiotic Stresses Downregulate Key Genes Involved in Nitrogen Uptake and Assimilation in Brassica juncea L.

    Directory of Open Access Journals (Sweden)

    Parul Goel

    Full Text Available Abiotic stresses such as salinity, drought and extreme temperatures affect nitrogen (N uptake and assimilation in plants. However, little is known about the regulation of N pathway genes at transcriptional level under abiotic stress conditions in Brassica juncea. In the present work, genes encoding nitrate transporters (NRT, ammonium transporters (AMT, nitrate reductase (NR, nitrite reductase (NiR, glutamine synthetase (GS, glutamate synthase (GOGAT, glutamate dehydrogenase (GDH, asparagines synthetase (ASN were cloned from Brassica juncea L. var. Varuna. The deduced protein sequences were analyzed to predict their subcellular localization, which confirmed localization of all the proteins in their respective cellular organelles. The protein sequences were also subjected to conserved domain identification, which confirmed presence of characteristic domains in all the proteins, indicating their putative functions. Moreover, expression of these genes was studied after 1h and 24h of salt (150 mM NaCl, osmotic (250 mM Mannitol, cold (4°C and heat (42°C stresses. Most of the genes encoding nitrate transporters and enzymes responsible for N assimilation and remobilization were found to be downregulated under abiotic stresses. The expression of BjAMT1.2, BjAMT2, BjGS1.1, BjGDH1 and BjASN2 was downregulated after 1hr, while expression of BjNRT1.1, BjNRT2.1, BjNiR1, BjAMT2, BjGDH1 and BjASN2 was downregulated after 24h of all the stress treatments. However, expression of BjNRT1.1, BjNRT1.5 and BjGDH2 was upregulated after 1h of all stress treatments, while no gene was found to be upregulated after 24h of stress treatments, commonly. These observations indicate that expression of most of the genes is adversely affected under abiotic stress conditions, particularly under prolonged stress exposure (24h, which may be one of the reasons of reduction in plant growth and development under abiotic stresses.

  10. Assessment of genetic diversity in Brassica juncea Brassicaceae genotypes using phenotypic differences and SSR markers

    Directory of Open Access Journals (Sweden)

    Vinu V.

    2013-12-01

    Full Text Available Las especies de mostaza del género Brassica representan uno de los cultivos de semillas oleaginosas más importantes en India, sin embargo, su diversidad genética es poco conocida. Para la utilización de genotipos en programas de cultivos resulta esencial un mayor conocimiento sobre este tema. Debido a ello, se evaluó la diversidad genética entre 44 genotipos de mostaza de la India Brassica juncea incluyendo variedades y líneas puras de diferentes zonas agro-climáticas de la India y algunos genotipos exóticos Australia, Polonia y China. Para ello, se utilizaron marcadores SSR específicos del genoma A y B y datos fenotípicos del rendimiento de 12 cosechas y sus características. De los 143 primers evaluados, 134 reportaron polimorfismo y un total de 355 alelos fueron amplificados. Se generaron dendrogramas a partir de los coeficientes de similitud de Jaccard y de disimilitud Manhattan, basados en un algoritmo de vinculación promedio UPGMA. Se utilizaron datos de marcadores genéticos y datos fenotípicos. Los genotipos se agruparon en cuatro grupos basados en las distancias genéticas. Ambos patrones de agrupamiento, tanto los basados en los coeficientes de similitud de Jaccard como los basados en los de disimilitud Manhattan, separaron independientemente los genotipos por su genealogía y origen, de una manera efectiva. El PCoA reveló que la agrupación de genotipos basada en datos de marcadores SSR, es más convincente que los datos fenotípicos, sin embargo, se observó que la correlación entre las matrices de distancia fenotípica y genética resultó muy baja r=0.11. Por lo tanto, para estudios de diversidad basados en marcadores moleculares es necesario realizar más pruebas. Los marcadores SSR constituyen herramientas más eficientes para discriminar entre genotipos de B. juncea, que las características cuantitativas.

  11. Taguchi Experimental Design for Optimization of Recombinant Human Growth Hormone Production in CHO Cell Lines and Comparing its Biological Activity with Prokaryotic Growth Hormone.

    Science.gov (United States)

    Aghili, Zahra Sadat; Zarkesh-Esfahani, Sayyed Hamid

    2017-09-12

    Growth hormone deficiency results in growth retardation in children and the GH deficiency syndrome in adults and they need to receive recombinant-GH in order to rectify the GH deficiency symptoms. Mammalian cells have become the favorite system for production of recombinant proteins for clinical application compared to prokaryotic systems because of their capability for appropriate protein folding, assembly, post-translational modification and proper signal. However, production level in mammalian cells is generally low compared to prokaryotic hosts. Taguchi has established orthogonal arrays to describe a large number of experimental situations mainly to reduce experimental errors and to enhance the efficiency and reproducibility of laboratory experiments.In the present study, rhGH was produced in CHO cells and production of rhGH was assessed using Dot blotting, western blotting and Elisa assay. For optimization of rhGH production in CHO cells using Taguchi method An M16 orthogonal experimental design was used to investigate four different culture components. The biological activity of rhGH was assessed using LHRE-TK-Luciferase reporter gene system in HEK-293 and compared to the biological activity of prokaryotic rhGH.A maximal productivity of rhGH was reached in the conditions of 1%DMSO, 1%glycerol, 25 µM ZnSO4 and 0 mM NaBu. Our findings indicate that control of culture conditions such as the addition of chemical components helps to develop an efficient large-scale and industrial process for the production of rhGH in CHO cells. Results of bioassay indicated that rhGH produced by CHO cells is able to induce GH-mediated intracellular cell signaling and showed higher bioactivity when compared to prokaryotic GH at the same concentrations. © Georg Thieme Verlag KG Stuttgart · New York.

  12. Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae, is permissive for AcMNPV infection and produces high levels of recombinant proteins

    Directory of Open Access Journals (Sweden)

    Zhang Sheng

    2010-07-01

    Full Text Available Abstract Background The insect cell line is a critical component in the production of recombinant proteins in the baculovirus expression system and new cell lines hold the promise of increasing both quantity and quality of protein production. Results Seventy cell lines were established by single-cell cloning from a primary culture of cells derived from eggs of the black witch moth (Ascalapha odorata; Lepidoptera, Noctuidae. Among 8 rapidly growing lines, cell line 38 (Ao38 was selected for further analysis, based on susceptibility to AcMNPV infection and production of secreted alkaline phosphatase (SEAP from a baculovirus expression vector. In comparisons with low-passage High Five (BTI-Tn-5B1-4 cells, infected Ao38 cells produced β-galactosidase and SEAP at levels higher (153% and 150%, respectively than those measured from High Five cells. Analysis of N-glycans of SEAP produced in Ao38 cells revealed two N-glycosylation sites and glycosylation patterns similar to those reported for High Five and Sf9 cells. Glycopeptide isoforms consisted of pauci- or oligomannose, with and without fucose on N-acetylglucosamine(s linked to asparagine residues. Estimates of Ao38 cell volume suggest that Ao38 cells are approximately 2.5× larger than Sf9 cells but only approximately 74% of the size of High Five cells. Ao38 cells were highly susceptible to AcMNPV infection, similar to infectivity of Sf9 cells. Production of infectious AcMNPV budded virions from Ao38 cells peaked at approximately 4.5 × 107 IU/ml, exceeding that from High Five cells while lower than that from Sf9 cells. Ao38 cells grew rapidly in stationary culture with a population doubling time of 20.2 hr, and Ao38 cells were readily adapted to serum-free medium (Sf-900III and to a suspension culture system. Analysis of Ao38 and a parental Ascalapha odorata cell line indicated that these lines were free of the alphanodavirus that was recently identified as an adventitious agent in High Five cell

  13. Effect of Salicylic Acid on Salinity-induced Changes in Brassica juncea

    Institute of Scientific and Technical Information of China (English)

    Mohammad Yusuf; Syed Aiman Hasan; Barket Ali; Shamsul Hayat; Qazi Fariduddin; Aqil Ahmad

    2008-01-01

    Seeds of Indian mustard (Brassica juncea (L.) Czern.et Coss.) were exposed to 0,50,100 and 150 mmol/L NaCl for 8h and seeds were sown in an earthen pot.These stressed seedlings were subsequently sprayed with 10 μrnol/L salicylic acid (SA) at 30 d and were sampled at 60 d to assess the changes in growth,photosynthesis and antioxidant enzymes.The seedlings raised from the seeds treated with NaCl had significantly reduced growth and the activities of carbonic anhydrase,nitrate reductase and photosynthesis,and the decrease was proportional to the increase in NaCl concentration.However,the antioxidant enzymes (catalase,peroxidase and superoxide dismutase) and proline content was enhanced in response to NaCl and/or SA treatment,where their interaction had an additive effect.Moreover,the toxic effects generated by the lower concentration of NaCl (50 mmol/L) were completely overcome by the application of SA.It was,therefore,concluded that SA ameliorated the stress generated by NaCl through the alleviated antioxidant system.

  14. Microbial production of succinic acid using crude and purified glycerol from a Crotalaria juncea based biorefinery

    Directory of Open Access Journals (Sweden)

    Suvra Sadhukhan

    2016-06-01

    Full Text Available Microbial conversion of crude and purified glycerol obtained in the process of biorefining Crotalaria juncea is carried out to produce succinic acid using Escherichia coli. Batch tests are performed for nine different substrate concentrations of commercial, purified and crude glycerol, in order to observe cell growth and substrate utilization rate. Inhibitory (Halden-Andrew, Aiba-Edward, Tessier type and Andrews as well as non-inhibitory (Monod, Moser and Tessier models are fitted to the relationship between specific growth rate and substrate concentration obtained from the growth curves. Considering the inhibition effect, Aiba-Edward model ranked 1 out of 7 in case of two samples and Haldane-Andrew model ranked 1 in case of one sample. Aiba-Edward model gave the best fitment for a large range of concentrations of all the three types of glycerol, crude, purified and laboratory grade. Maximum production of succinic acid is obtained from commercial glycerol at pH 7 and 37.5 °C.

  15. A computational study of the aerodynamic forces and power requirements of dragonfly Aeschna juncea hovering

    CERN Document Server

    Sun, M; Sun, Mao; Lan, Shi Long

    2004-01-01

    Aerodynamic force generation and mechanical power requirements of a dragonfly (Aeschna juncea) in hovering flight are studied. The method of numerically solving the Navier-Stokes equations in moving overset grids is used. There are two large vertical force peaks in one flapping cycle. One is in the first half of the cycle, which is mainly due to the hindwings in their downstroke; the other is in the second half of the cycle, which is mainly due to the forewings in their downstroke. Hovering with a large stroke plane angle, the dragonfly uses drag as a major source for its weight supporting force (approximately 65% of the total vertical force is contributed by the drag and 35% by the lift of the wings). The vertical force coefficient of a wing is twice as large as the quasi-steady value. The interaction between the fore- and hindwings is not very strong and is detrimental to the vertical force generation. Compared with the case of a single wing in the same motion, the interaction effect reduces the vertical fo...

  16. The intensity of tyrosine nitration is associated with selenite and selenate toxicity in Brassica juncea L.

    Science.gov (United States)

    Molnár, Árpád; Feigl, Gábor; Trifán, Vanda; Ördög, Attila; Szőllősi, Réka; Erdei, László; Kolbert, Zsuzsanna

    2017-08-21

    Selenium phytotoxicity involves processes like reactive nitrogen species overproduction and nitrosative protein modifications. This study evaluates the toxicity of two selenium forms (selenite and selenate at 0µM, 20µM, 50µM and 100µM concentrations) and its correlation with protein tyrosine nitration in the organs of hydroponically grown Indian mustard (Brassica juncea L.). Selenate treatment resulted in large selenium accumulation in both Brassica organs, while selenite showed slight root-to-shoot translocation resulting in a much lower selenium accumulation in the shoot. Shoot and root growth inhibition and cell viability loss revealed that Brassica tolerates selenate better than selenite. Results also show that relative high amounts of selenium are able to accumulate in Brassica leaves without obvious visible symptoms such as chlorosis or necrosis. The more severe phytotoxicity of selenite was accompanied by more intense protein tyrosine nitration as well as alterations in nitration pattern suggesting a correlation between the degree of Se forms-induced toxicities and nitroproteome size, composition in Brassica organs. These results imply the possibility of considering protein tyrosine nitration as novel biomarker of selenium phytotoxicity, which could help the evaluation of asymptomatic selenium stress of plants. Copyright © 2017. Published by Elsevier Inc.

  17. The EDTA Amendment in Phytoextraction of (134)Cs From Soil by Indian Mustard (Brassica juncea).

    Science.gov (United States)

    Tjahaja, Poppy Intan; Sukmabuana, Putu; Roosmini, Dwina

    2015-01-01

    Soil contamination with radiocaesium is a significant problem at any countries when a nuclear accident occurred. Recently, phytoextraction technique is developed to remediate the contaminated environment. However, the application is limited by the availability of the contaminant for root uptake. Therefore, a green house trial experiment of soil amendment with ethylene diamine tetraacetic acid (EDTA) has been conducted to examine (134)Cs availability for root uptake. Two groups of Indian mustard (Brassica juncea) were cultivated in (134)Cs contaminated soil. The soil in the first group was treated with EDTA amendment, while the other was not. Plant growth was observed gravimetrically and the (134)Cs concentration in soil as well as plants were determined using gamma spectrometry. The plant uptake capacity was determined as transfer factor (Fv), and the Fv values of 0.22 ± 0.0786 and 0.12 ± 0.039 were obtained for the soil treated with and without EDTA amendment, respectively. The phytoextraction efficiency of the plant cultivated in (134)Cs contaminated soil both with and without EDTA amendment was low. The EDTA amendment to the soil seems to enhance the (134)Cs availability for root uptake of Indian mustard and can still be considered to assist the field phytoremediation of contaminated soil.

  18. Isolation and functional analysis of a Brassica juncea gene encoding a component of auxin efflux carrier

    Institute of Scientific and Technical Information of China (English)

    WEI; MIN; NI; XIAO; YA; CHEN; ZHI; HONG; XU; HONG; WEI; XUE

    2002-01-01

    Polar auxin transport plays a divergent role in plant growth and developmental processes including rootand embryo development, vascular pattern formation and cell elongation. Recently isolated Arabidopsispin gene family was believed to encode a component of auxin efflux carrier (Galweiler et al, 1998). Basedon the Arabidopsis pin1 sequence we have isolated a Brassica juncea cDNA (designated Bjpinl), whichencoded a 70-kDa putative auxin efflux carrier. Deduced BjPIN1 shared 65% identities at protein level withAtPIN1 and was highly homologous to other putative PIN proteins of Arabidopsis (with highest homologyto AtPIN3). Hydrophobic analysis showed similar structures between BjPIN1 and AtPIN proteins. Presenceof 6 exons (varying in size between 65 bp and 1229 bp) and 5 introns (sizes between 89 bp and 463 bp)in the genomic fragment was revealed by comparing the genomic and cDNA sequences. Northern blotanalysis indicated that Bjpinl was expressed in most of the tissues tested, with a relatively higher levelof transcript in flowers and a lower level in root tissues. Promoter-reporter gene fusion studies furtherrevealed the expression of Bjpinl in the mature pollen grains, young seeds, root tip, leaf vascular tissue andtrace bundle, stem epidermis, cortex and vascular cells. BjPIN1 was localized on the plasma membraneas demonstrated through fusion expression of green fluorescent protein (GFP). Auxin efflux carrier activitywas elevated in transgenic Arabidopsis expressing BjPIN1.

  19. Biorefinery process for protein extraction from oriental mustard (Brassica juncea (L.) Czern.) using ethanol stillage.

    Science.gov (United States)

    Ratanapariyanuch, Kornsulee; Tyler, Robert T; Shim, Youn Young; Reaney, Martin Jt

    2012-01-12

    Large volumes of treated process water are required for protein extraction. Evaporation of this water contributes greatly to the energy consumed in enriching protein products. Thin stillage remaining from ethanol production is available in large volumes and may be suitable for extracting protein rich materials. In this work protein was extracted from ground defatted oriental mustard (Brassica juncea (L.) Czern.) meal using thin stillage. Protein extraction efficiency was studied at pHs between 7.6 and 10.4 and salt concentrations between 3.4 × 10-2 and 1.2 M. The optimum extraction efficiency was pH 10.0 and 1.0 M NaCl. Napin and cruciferin were the most prevalent proteins in the isolate. The isolate exhibited high in vitro digestibility (74.9 ± 0.80%) and lysine content (5.2 ± 0.2 g/100 g of protein). No differences in the efficiency of extraction, SDS-PAGE profile, digestibility, lysine availability, or amino acid composition were observed between protein extracted with thin stillage and that extracted with NaCl solution. The use of thin stillage, in lieu of water, for protein extraction would decrease the energy requirements and waste disposal costs of the protein isolation and biofuel production processes.

  20. Microbial production of succinic acid using crude and purified glycerol from a Crotalaria juncea based biorefinery.

    Science.gov (United States)

    Sadhukhan, Suvra; Villa, Raffaella; Sarkar, Ujjaini

    2016-06-01

    Microbial conversion of crude and purified glycerol obtained in the process of biorefining Crotalaria juncea is carried out to produce succinic acid using Escherichia coli. Batch tests are performed for nine different substrate concentrations of commercial, purified and crude glycerol, in order to observe cell growth and substrate utilization rate. Inhibitory (Halden-Andrew, Aiba-Edward, Tessier type and Andrews) as well as non-inhibitory (Monod, Moser and Tessier) models are fitted to the relationship between specific growth rate and substrate concentration obtained from the growth curves. Considering the inhibition effect, Aiba-Edward model ranked 1 out of 7 in case of two samples and Haldane-Andrew model ranked 1 in case of one sample. Aiba-Edward model gave the best fitment for a large range of concentrations of all the three types of glycerol, crude, purified and laboratory grade. Maximum production of succinic acid is obtained from commercial glycerol at pH 7 and 37.5 °C.

  1. The influence of different hydroponic conditions on thorium uptake by Brassica juncea var. foliosa.

    Science.gov (United States)

    Wang, Dingna; Zhou, Sai; Liu, Li; Du, Liang; Wang, Jianmei; Huang, Zhenling; Ma, Lijian; Ding, Songdong; Zhang, Dong; Wang, Ruibing; Jin, Yongdong; Xia, Chuanqin

    2015-05-01

    The effects of different hydroponic conditions (such as concentration of thorium (Th), pH, carbonate, phosphate, organic acids, and cations) on thorium uptake by Brassica juncea var. foliosa were evaluated. The results showed that acidic cultivation solutions enhanced thorium accumulation in the plants. Phosphate and carbonate inhibited thorium accumulation in plants, possibly due to the formation of Th(HPO4)(2+), Th(HPO4)2, or Th(OH)3CO3 (-) with Th(4+), which was disadvantageous for thorium uptake in the plants. Organic aids (citric acid, oxalic acid, lactic acid) inhibited thorium accumulation in roots and increased thorium content in the shoots, which suggested that the thorium-organic complexes did not remain in the roots and were beneficial for thorium transfer from the roots to the shoots. Among three cations (such as calcium ion (Ca(2+)), ferrous ion (Fe(2+)), and zinc ion (Zn(2+))) in hydroponic media, Zn(2+) had no significant influence on thorium accumulation in the roots, Fe(2+) inhibited thorium accumulation in the roots, and Ca(2+) was found to facilitate thorium accumulation in the roots to a certain extent. This research will help to further understand the mechanism of thorium uptake in plants.

  2. Multitrait plant growth promoting (PGP) rhizobacterial isolates from Brassica juncea rhizosphere : Keratin degradation and growth promotion.

    Science.gov (United States)

    Anwar, Mohmmad Shahbaz; Siddique, Mohammad Tahir; Verma, Amit; Rao, Yalaga Rama; Nailwal, Tapan; Ansari, Mohammad; Pande, Veena

    2014-01-01

    Plant growth promoting (PGP) rhizobacteria, a beneficial microbe colonizing plant roots, enhanced crop productivity and offers an attractive way to replace chemical fertilizers, pesticides, and supplements. The keratinous waste which comprises feathers, hairs, nails, skin and wool creates problem of solid waste management due to presence of highly recalcitrant keratin. The multi traits rhizobacteria effective to remove both keratine from the environment by producing keratinase enzyme and to eradicate the chemical fertilizer by providing different PGP activity is novel achievement. In the present study, the effective PM2 strain of PGPR was isolated from rhizospheric soil of mustard (Brassica juncea) field, Pantnagar and they were identified on the basis of different biochemical tests as belonging to Bacillus genera. Different plant growth promoting activity, feather degradation and keratinolytic activity was performed and found very effective toward all the parameters. Furthermore, the efficient strain PM2 was identified on the basis of 16s rRNA sequencing and confirmed as Bacillus cereus. The strain PM2 might be used efficiently for keratinous waste management and PGP activity. Therefore, the present study suggests that Bacillus cereus have multi traits activity which extremely useful for different PGP activity and biotechnological process involving keratin hydrolysis, feather biodegradation or in the leather industry.

  3. The effect of Bt-transgene introgression on plant growth and reproduction in wild Brassica juncea.

    Science.gov (United States)

    Liu, Yong-Bo; Darmency, Henry; Stewart, C Neal; Wei, Wei; Tang, Zhi-Xi; Ma, Ke-Ping

    2015-06-01

    This study aims to investigate the relative plant growth and reproduction of insect-resistant and susceptible plants following the introgression of an insect-resistance Bt-transgene from Brassica napus, oilseed rape, to wild Brassica juncea. The second backcrossed generation (BC2) from a single backcross family was grown in pure and mixed stands of Bt-transgenic and non-transgenic siblings under two insect treatments. Various proportions of Bt-transgenic plants were employed in mixed stands to study the interaction between resistant and susceptible plants. In the pure stands, Bt-transgenic BC2 plants performed better than non-transgenic plants with or without insect treatments. In mixed stands, Bt-transgenic BC2 plants produced fewer seeds than their non-Bt counterparts at low proportions of Bt-transgenic BC2 plants in the absence of insects. Reproductive allocation of non-transgenic plants marginally increased with increasing proportions of Bt-transgenic plants under herbivore pressure, which resulted in increased total biomass and seed production per stand. The results showed that the growth of non-transgenic plants was protected by Bt-transgenic plants under herbivore pressure. The Bt-transgene might not be advantageous in mixed stands of backcrossed hybrids; thus transgene introgression would not be facilitated when herbivorous insects are not present. However, a relatively large initial population of Bt-transgenic plants might result in transgene persistence when target herbivores are present.

  4. Growth response modulation by putrescine in Indian mustard Brassica juncea L. under multiple stress.

    Science.gov (United States)

    Lakra, Nita; Tomar, Pushpa C; Mishra, S N

    2016-04-01

    Plants, in general, are put to various kinds of stress, biotic and abiotic, both natural and manmade. Infestation by insect pests and diseases, and extreme conditions such as salinity, temperature, etc., as well as heavy metal contamination affect their growth performance. Here, we studied the impact of salinity and heavy metal pollution on the growth performance of Indian Mustard Brassica juncea L. and its amelioration by the diamine, putrescine, a known media supplement. We evaluated the putrescine (Put) modulation potential on multiple stress effect in 7-day old Indian mustard. The germination, seedlings length and photosynthetic pigments decline under salinity and metal (Cd/Pb) stress condition, alone or in combination, were checked by putrescine. The stress induced increase in root-shoot ratio, RNA and total amino acids content, as well as Na⁺/K⁺ ratio in leaf tissues were also comparatively less. The increased endogenous Cd/Pb accumulation in plants exposed to either metal further elevated under salinity was also found decelerated. However, the multiple stressed seedlings showed increase in glutathione content, which was further elevated with putrescine application. The increase in protein contents in leaf under single or combined stresses in the presence of putrescine could be a qualitative change. The differential changes in parameters examined here resulted in improved growth (> 10%) suggests stress mitigation by the putrescine up to an extent.

  5. Multitrait plant growth promoting (PGP) rhizobacterial isolates from Brassica juncea rhizosphere

    Science.gov (United States)

    Anwar, Mohmmad Shahbaz; Siddique, Mohammad Tahir; Verma, Amit; Rao, Yalaga Rama; Nailwal, Tapan; Ansari, Mohammad; Pande, Veena

    2014-01-01

    Plant growth promoting (PGP) rhizobacteria, a beneficial microbe colonizing plant roots, enhanced crop productivity and offers an attractive way to replace chemical fertilizers, pesticides, and supplements. The keratinous waste which comprises feathers, hairs, nails, skin and wool creates problem of solid waste management due to presence of highly recalcitrant keratin. The multi traits rhizobacteria effective to remove both keratine from the environment by producing keratinase enzyme and to eradicate the chemical fertilizer by providing different PGP activity is novel achievement. In the present study, the effective PM2 strain of PGPR was isolated from rhizospheric soil of mustard (Brassica juncea) field, Pantnagar and they were identified on the basis of different biochemical tests as belonging to Bacillus genera. Different plant growth promoting activity, feather degradation and keratinolytic activity was performed and found very effective toward all the parameters. Furthermore, the efficient strain PM2 was identified on the basis of 16s rRNA sequencing and confirmed as Bacillus cereus. The strain PM2 might be used efficiently for keratinous waste management and PGP activity. Therefore, the present study suggests that Bacillus cereus have multi traits activity which extremely useful for different PGP activity and biotechnological process involving keratin hydrolysis, feather biodegradation or in the leather industry. PMID:24778758

  6. Lithium, Vanadium and Chromium Uptake Ability of Brassica juncea from Lithium Mine Tailings.

    Science.gov (United States)

    Elektorowicz, M; Keropian, Z

    2015-01-01

    The potential for phytoremediation and phytostabilization of lithium in lieu with vanadium and chromium on a formulated acidic heterogeneous growth media engineered around lithium mine tailings, was investigated in four phases: (1) overall efficiency of the removal of the three metals, (2) bioaccumulation ratios of the three metals, (3) overall relative growth rate, and (4) translocation index of the three metals in the physiology of the hyperaccumulator plant. A pot study was conducted to assess the suitability of Brassica juncea (Indian mustard) in a phytoremediation process whereby it was lingered for eighty-six days under homogeneous growth conditions and irrigated bidaily with organic fertilizer amended with LiCl. A post harvest data analysis was achieved through ashing and the implementation of cold digestion procedure in a concentrated hydrochloric acidic matrix. In physiological efficiency parameters, the hyperaccumulator plant was twice as able to phytostabilize chromium and four times was able to phytostabilize vanadium in comparison to lithium. Moreover, it was extremely efficient in translocating and accumulating lithium inside its upper physiological sites, more so than chromium and vanadium, thereby demonstrating Indian mustard, as a hyperaccumulator plant, for phytoextraction and phytostabilization in an acidic heterogeneous rhizosphere, with an extremely low relative growth rate.

  7. Identification of genomic region controlling resistance to aflatoxin contamination in a peanut recombinant inbred line population (Tifrunner XGT-C20)

    Science.gov (United States)

    In peanut, limited genetic variation for disease resistance is available in breeding programs necessitating the identification of stable resistance sources for use in cultivar development. ‘Tifrunner’ is a runner cultivar while ‘GT-C20’ is a Spanish-type breeding line with resistance to aflatoxin co...

  8. Genetic mapping of major-effect seed dormancy quantitative trait loci on chromosome 2B using recombinant substitution lines in tetraploid wheat

    Science.gov (United States)

    Durum wheat cultivars can benefit from having some level of seed dormancy to help reduce seed damage and lower grain quality caused by pre-harvest sprouting (PHS) occurring during wet harvesting conditions. Previously a single chromosome substitution line carrying chromosome 2B of wild emmer in the...

  9. Soil organisms associated to the weed suppressant Crotalaria juncea (fabaceae and its importance as a refuge for natural enemies Organismos de solo associados à supressora de plantas daninhas Crotalaria juncea (fabaceae e sua importância como refúgio para inimigos naturais

    Directory of Open Access Journals (Sweden)

    W.S Tavares

    2011-09-01

    Full Text Available Soil organisms play an important role in organic crops of Crotalaria juncea (Fabaceae and are associated with the natural conservation of the environment. The present study was aimed to investigate the population of soil organisms in the organic culture of C. juncea, as well as its importance as a refuge for natural enemies. Dalbulus maidis (Hemiptera: Cicadellidae, Diabrotica sp. (Coleoptera: Chrysomelidae, Doru luteipes (Dermaptera: Forficulidae, Gryllus assimilis (Orthoptera: Gryllidae, Lagria villosa (Coleoptera: Lagriidae, Melanotus sp. (Coleoptera: Elateridae, Meloidogyne incognita (Tylenchida: Heteroderidae, Nephila clavipes (Araneae: Nephilidae, Orius insidiosus (Hemiptera: Anthocoridae, Pheidole sp. (Hymenoptera: Myrmicidae, Phyllophaga sp. (Coleoptera: Scarabeidae, Procornitermes sp. (Isoptera: Termitidae, Solenopsis sp. (Hymenoptera: Formicidae, and Utetheisa ornatrix (Lepidoptera: Arctiidae were identified in C. juncea. The organisms that were found during a 3-month period in 144 trenches in C. juncea were pest species (84.47% and natural enemies (15.53% as well. Natural enemies had an average of 11.89 individuals per 1.08 m³ of soil cultivated with C. juncea. The abundance of organisms in the pod stage (5.49% of C. juncea was lower than that in the vegetative (83.50% and flowering (11.01% stages. Crotalaria juncea plants can be used as part of a crop system for Integrated Pest Management.Organismos de solo desempenham um importante papel em cultivos orgânicos de Crotalaria juncea (Fabaceae e estão associados com a conservação natural do ambiente. O presente estudo teve como objetivo investigar a população de organismos de solo no cultivo orgânico de C. juncea, bem como sua importância como um refúgio para inimigos naturais. Dalbulus maidis (Hemiptera: Cicadellidae, Diabrotica sp. (Coleoptera: Chrysomelidae, Doru luteipes (Dermaptera: Forficulidae, Gryllus assimilis (Orthoptera: Gryllidae, Lagria villosa (Coleoptera

  10. Fitorremediação de solo contaminado com sulfentrazone em função da densidade populacional de Canavalia ensiformis e Crotalaria juncea.

    OpenAIRE

    Araujo,M.F.

    2012-01-01

    Objetivou-se neste trabalho avaliar a influência da densidade populacional de feijão-de-porco (Canavalia ensiformis) e crotalária juncea (Crotalaria juncea) sobre a fitorremediação de solo contaminado com o herbicida sulfentrazone. O experimento foi conduzido em casa-de-vegetação, utilizando-se vasos com solo coletado do horizonte A de um Argilosso Amarelo arenoso, no delineamento em blocos casualizados, em esquema fatorial 4 x 3, com quatro repetições. Os tratamentos foram compostos pela com...

  11. SUSTAINABILITY EFFECTS OF Crotalaria juncea L. AND Crotalaria spectabilis ROTH ON SOIL FERTILITY AND SOIL CONSERVATION

    Science.gov (United States)

    László, Márton, ,, Dr.

    2010-05-01

    Sustainable agriculture is defined as the successful management of resources for agriculture to satisfy changing human needs while maintaining or enhancing the quality of the environment and conserving natural resources. A sustained increase of agricultural production becomes a great possibility for international community. In this process a green manure crops application for example crotalaria get a new chance for improvement process on soil fertility and soil conservation. Field experiment was carried out on a calcareous chernozem soil (Experiment station Nagyhörcsök of RISSAC-HAS) in partly of experiment series (3 years) at Hungary in 1998. The soil with about 20% clay, 3% humus, 5% CaCO3 in its ploughed layer. To ensure a sufficient macro and micronutrient supply in the whole experiment, 100 kg N, 100 kg P2O5 and 100 kg K2O were given hectare. The Crotalaria juncea L. and Crotalaria spectabilis ROTH were applied with 2 replications. Each plot has an area of 45 m2 with 230-230 individual plants. In vegetation grown period were measured green and dry matter yield. The soil and plant samples were analysed for the macro and microelements contents. The main results achieved in 1998 are summarized as follows: 1. The green matter yield at before flowering reached 63.8 t ha-1 in case of Crotalaria juncea L. 2. Total dry matter yield at harvest (without roots) fluctuated between 9.6 and 17.0 t ha-1, depending on the crotalaria species. 3. The average of element concentration (including stems, leaves of Crotalaria juncea L. and Crotalaria spectabilis ROTH) before flowering reached to 3.2 % N, 2.3 % Ca, 1.3 % K, 0.39 % Mg, 0.22 % P and 0.24 % S. The content of Al and Fe total 14 - 25, while that of Sr, Mn, Na, B and Ba 2 - 6 ppm in dry matter. The Zn, Cu, Mo, Cr, Se, Ni, As, Pb, Cd and Co concentration did not reach here the value of 1 ppm. 4. The average of biological activated element uptake (including stems, leaves of Crotalaria juncea L. and Crotalaria spectabilis

  12. Effect of microwave treatment on the efficacy of expeller pressing of Brassica napus rapeseed and Brassica juncea mustard seeds.

    Science.gov (United States)

    Niu, Yanxing; Rogiewicz, Anna; Wan, Chuyun; Guo, Mian; Huang, Fenghong; Slominski, Bogdan A

    2015-04-01

    A study was conducted to evaluate the effect of microwave heating on the efficacy of expeller pressing of rapeseed and mustard seed and the composition of expeller meals in two types of Brassica napus rapeseed (intermediate- and low-glucosinolate) and in Brassica juncea mustard (high-glucosinolate). Following microwave treatment, the microstructure of rapeseed using transmission electron microscopy showed a significant disappearance of oil bodies and myrosin cells. After 6 min of microwave heating (400 g, 800 W), the oil content of rapeseed expeller meal decreased from 44.9 to 13.5% for intermediate-glucosinolate B. napus rapeseed, from 42.6 to 11.3% for low-glucosinolate B. napus rapeseed, and from 44.4 to 14.1% for B. juncea mustard. The latter values were much lower than the oil contents of the corresponding expeller meals derived from the unheated seeds (i.e., 26.6, 22.6, and 29.8%, respectively). Neutral detergent fiber (NDF) contents showed no differences except for the expeller meal from the intermediate-glucosinolate B. napus rapeseed, which increased from 22.7 to 29.2% after 6 min of microwave heating. Microwave treatment for 4 and 5 min effectively inactivated myrosinase enzyme of intermediate-glucosinolate B. napus rapeseed and B. juncea mustard seed, respectively. In low-glucosinolate B. napus rapeseed the enzyme appeared to be more heat stable, with some activity being present after 6 min of microwave heating. Myrosinase enzyme inactivation had a profound effect on the glucosinolate content of expeller meals and prevented their hydrolysis to toxic breakdown products during the expelling process. It appeared evident from this study that microwave heating for 6 min was an effective method of producing expeller meal without toxic glucosinolate breakdown products while at the same time facilitating high yield of oil during the expelling process.

  13. Detecting the cosmological recombination signal from space

    CERN Document Server

    Desjacques, Vincent; Silk, Joseph; de Bernardis, Francesco; Doré, Olivier

    2015-01-01

    Spectral distortions of the CMB have recently experienced an increased interest. One of the inevitable distortion signals of our cosmological concordance model is created by the cosmological recombination process, just a little before photons last scatter at redshift $z\\simeq 1100$. These cosmological recombination lines, emitted by the hydrogen and helium plasma, should still be observable as tiny deviation from the CMB blackbody spectrum in the cm--dm spectral bands. In this paper, we present a forecast for the detectability of the recombination signal with future satellite experiments. We argue that serious consideration for future CMB experiments in space should be given to probing spectral distortions and, in particular, the recombination line signals. The cosmological recombination radiation not only allows determination of standard cosmological parameters, but also provides a direct observational confirmation for one of the key ingredients of our cosmological model: the cosmological recombination histo...

  14. Effects of phosphate and thiosulphate on arsenic accumulation in Brassica juncea plants grown in soil and in hydroponic culture

    Science.gov (United States)

    Pezzarossa, Beatrice; Petruzzelli, Gianniantonio; Grifoni, Martina; Rosellini, Irene; Malagoli, Mario; Schiavon, Michela

    2013-04-01

    Arsenic is recognised as a toxic metalloid and a strong pollutant in soils of many countries. Thus, the reclamation of contaminated areas is fundamental in order to protect both human health and agricultural production. This study is focused on the assisted phytoextraction, a technology for reclaiming polluted soils that takes advantage of the capability of some plants to extract inorganic elements from soils with the aid of additive agents. The nutrients phosphorus, as phosphate, and sulphur, as thiosulphate, can compete with the form more oxidised of arsenic, both in soil and plant. This study examined the capability of thiosulphate (Th) and phosphate (Ph) to promote the release of As from soil surfaces in order to improve the phytoavailability and thus the absorption of As by Brassica juncea plants. In the first experiment B. juncea plants were grown on a soil that had been sampled from an industrial area strongly contaminated by As (790 mg As kg-1 soil). The second experiment was carried out in hydroponics where As has been added at a concentration (100 microM) similar to the As available concentration measured in soil. In both trials ammonium thiosulphate (at the concentration of 0.27 M in soil, and 400 microM in hydroponics) and potassium hydrogen phosphate (at the concentration of 0.05 M in soil, and 112 microM in hydroponics) were added. The biomass of B. juncea was determined and the accumulation of P, S and As in root and in the above-ground tissues have been analyzed. Our results showed that thiosulphate and phosphate acted either as nutrients and detoxifying agents, due to the stimulation of plant defensive systems, and influenced either the biomass production and the As accumulation in plant tissues. In the plants grown in soil, As accumulated at higher levels in the above-ground part than in the roots and the addition of Th induced a higher biomass production and a higher total As accumulation (concentration x biomass) in the above-ground tissues

  15. Expression of recombinant antibodies.

    Science.gov (United States)

    Frenzel, André; Hust, Michael; Schirrmann, Thomas

    2013-01-01

    Recombinant antibodies are highly specific detection probes in research, diagnostics, and have emerged over the last two decades as the fastest growing class of therapeutic proteins. Antibody generation has been dramatically accelerated by in vitro selection systems, particularly phage display. An increasing variety of recombinant production systems have been developed, ranging from Gram-negative and positive bacteria, yeasts and filamentous fungi, insect cell lines, mammalian cells to transgenic plants and animals. Currently, almost all therapeutic antibodies are still produced in mammalian cell lines in order to reduce the risk of immunogenicity due to altered, non-human glycosylation patterns. However, recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with "human-like" post-translational modifications. Furthermore, smaller antibody fragments including bispecific antibodies without any glycosylation are successfully produced in bacteria and have advanced to clinical testing. The first therapeutic antibody products from a non-mammalian source can be expected in coming next years. In this review, we focus on current antibody production systems including their usability for different applications.

  16. Thermal requirement of indian mustard (Brassica juncea) at different phonological stages under late sown condition.

    Science.gov (United States)

    Singh, Manoj Pratap; Lallu; Singh, N B

    2014-01-01

    Indian mustard [Brassica juncea (L.) Czern & Coss.] is a long day plant, which requires fairly cool climatic condition during growth and development for obtaining better seed yield. Various workers have correlated crop growth and development with energy requirement parameters, such as growing degree days (GDD), photo-thermal unit (PTU), helios thermal unit (HTU), photo-thermal index (PTI) and heat use efficiency (HUE). Therefore, GDD requirement for different phenological stages of 22 newly developed Indian mustard varieties was studies during winter (rabi) seasons of 2009-10 and 2010-11 at student instructional farm of C.S. Azad University of Agriculture and Technology, Kanpur, (Utter Pradesh). Study revealed that RH-8814, NRCDR-02 and BPR-549-9 recorded higher GDD (1703.0, 1662.9 and 1648.0), PTU (19129.8, 18694.2 and 18379.8), HTU (11397.7, 11072.2 and 10876.0), PTI (13.25, 13.14 and 13.08) and HUE (4.11, 3.84 and 3.71) at physiological maturity, while higher HUE was recorded (9.62, 8.99 and 8.91 kg ha(-1) degrees-day) at days after sowing (DAS) to 50 % flowering. On the basis of study mustard genotypes RH-8814, NRCDR-02 and BPR-549-9 were identified as most heat-tolerant, as they maintained higher values for energy related parameters. Seed yield was highly positively correlated with GDD (r = 0.61, 0.65 and 0.75), PTU (r = 0.66, 0.64 and 0.74), HTU(r = 0.79, 0.68 and 0.73) at the above these three phenological stages, while negatively correlated with PTI at anthesis and at maturity. Hence, these parents could be used in crossing programme for achieving further improvement.

  17. In-vitro fermentation characteristics and methane reduction potential of mustard cake (Brassica juncea L.

    Directory of Open Access Journals (Sweden)

    S. M. Durge

    2016-10-01

    Full Text Available Aim: To assess the effect of mustard cake (Brassica juncea L. levels in concentrate mixtures and in composite feed mixtures (CFMs on in-vitro fermentation characteristics and methane production. Materials and Methods: Five concentrate mixtures were prepared with containing 30% oil cake, where linseed cake was replaced by mustard cake at the rate of 0%, 7.5%, 15.0%, 22.5%, and 30% in concentrate mixture. Mustard cake contained glucosinolate 72.58 μmol/g oil free dry matter (DM and contents in diet were 0, 5.4, 10.9, 16.3, and 21.8 μmol/g of concentrate mixture, respectively. Concentrate mixture containing 15.0% mustard cake was found to produced minimum methane which was then used for the preparation of CFM containing 0%, 25%, 50%, and 75% levels with gram straw. Result: Increased levels of mustard cake in concentrate mixtures had a linear decrease (p<0.05 in the total gas production, and the 15% inclusion showed lowest methane concentration (quadratic, p<0.01. The degradability of DM and organic matter (OM of concentrate mixtures did not change, however, pH and NH3-N concentrations of the fermentation medium showed linear (p<0.05 reductions with increased mustard cake levels. Increased levels of 15% mustard cake containing concentrate mixture in CFMs exhibited a trend (p=0.052 of increased gas production, whereas methane concentration in total gas, methane produced and degradability of DM and OM were also displayed a linear increase (p<0.05. However, the pH, NH3-N, and total volatile fatty acid levels decreased linearly (p<0.05 with increased levels of concentrate in CFMs. Conclusion: Reduction in methane production was evidenced with the inclusion of mustard cake in concentrate mixture at 15% level, and the CFMs with 25% concentrate, which contained 15% mustard cake, exhibited an improved fermentation and reduced methane production.

  18. Effects of phosphate and thiosulphate on arsenic accumulation in the species Brassica juncea.

    Science.gov (United States)

    Grifoni, Martina; Schiavon, Michela; Pezzarossa, Beatrice; Petruzzelli, Gianniantonio; Malagoli, Mario

    2015-02-01

    Arsenic (As) is recognized as a toxic pollutant in soils of many countries. Since phosphorus (P) and sulphur (S) can influence arsenic mobility and bioavailability, as well as the plant tolerance to As, phytoremediation techniques employed to clean-up As-contaminated areas should consider the interaction between As and these two nutrients. In this study, the bioavailability and accumulation of arsenate in the species Brassica juncea were compared between soil system and hydroponics in relation to P and S concentration of the growth substrate. In one case, plants were grown in pots filled with soil containing 878 mg As kg(-1). The addition of P to soil resulted in increased As desorption and significantly higher As accumulation in plants, with no effect on growth. The absence of toxic effects on plants was likely due to high S in soil, which could efficiently mitigate metal toxicity. In the hydroponic experiment, plants were grown with different combinations of As (0 or 100 μM) and P (56 or 112 μM). S at 400 μM was also added to the nutrient solution of control (-As) and As-treated plants, either individually or in combination with P. The addition of P reduced As uptake by plants, while high S resulted in higher As accumulation and lower P content. These results suggest that S can influence the interaction between P and As for the uptake by plants. The combined increase of P and S in the nutrient solution did not lead to higher accumulation of As, but enhanced As translocation from the root to the shoot. This aspect is of relevance for the phytoremediation of As-contaminated sites.

  19. Identification of candidate genes associated with cell wall digestibility and eQTL (expression quantitative trait loci analysis in a Flint × Flint maize recombinant inbred line population

    Directory of Open Access Journals (Sweden)

    Wenzel Gerhard

    2007-01-01

    Full Text Available Abstract Background Cell-wall digestibility is the major target for improving the feeding value of forage maize. An understanding of the molecular basis for cell-wall digestibility is crucial towards breeding of highly digestible maize. Results 865 candidate ESTs for cell-wall digestibility were selected according to the analysis of expression profiles in 1 three sets of brown-midrib isogenic lines in the genetic background of inbreds 1332 (1332 and 1332 bm3, 5361 (5361 and 5361 bm3, and F2 (F2, F2 bm1, F2 bm2, and F2 bm3, 2 the contrasting extreme lines of FD (Flint × Dent, AS08 × AS 06, DD1 (Dent × Dent, AS11 × AS09, and DD2 (Dent × Dent, AS29 × AS30 mapping populations, and 3 two contrasting isogenic inbreds, AS20 and AS21. Out of those, 439 ESTs were assembled on our "Forage Quality Array", a small microarray specific for cell wall digestibility related experiments. Transcript profiles of 40 lines of a Flint × Flint population were monitored using the Forage Quality Array, which were contrasting for cell wall digestibility. Using t-tests (p Conclusion 102 candidate genes for cell-wall digestibility were validated by genetical genomics approach. Although the cDNA array highlights gene types (the tested gene and any close family members, trans-acting factors or metabolic bottlenecks seem to play the major role in controlling heritable variation of gene expression related to cell-wall digestibility, since no in silico mapped ESTs were in the same location as their own eQTL. Transcriptional variation was generally found to be oligogenic rather than monogenic inherited due to only 26% ESTs detected a single eQTL in the present study. One eQTL hotspot was co-localized with cell wall digestibility related QTL cluster on bins 3.05, implying that in this case the gene(s underlying QTL and eQTL are identical. As the field of genetical genomics develops, it is expected to significantly improve our knowledge about complex traits, such as cell

  20. Establishment of a mammalian cell line suitable for industrial production of recombinant protein using mutations induced by high-energy beam radiation.

    Science.gov (United States)

    Chida, Yasuhito; Takagi, Keiichi; Terada, Satoshi

    2013-12-01

    Mammalian cells are extensively used for production of biopharmaceuticals. Most cells used in industry have infinite proliferative capacity, which provides a high number of cells and corresponding productivity. However, infinite cells will continue to multiply even after cell density reaches sufficient levels. This excess proliferation aggravates the culture environment and induces low productivity. Therefore, after cell density reaches sufficient levels, downregulation of proliferation would prevent such aggravation and extend the culture period and improve productivity. To realize such suitable proliferation, we aimed to establish a novel cell line whose proliferation was spontaneously downregulated after reaching a sufficient population level. Mutagenesis using high-energy beam irradiation was used. CHO-DP12 cells were irradiated with 2.5 Gy X-rays and screened with hydroxyurea and 5-fluorouracil to eliminate any cells multiplying after confluence and to concentrate desired mutants. One clone was established and named CHO-M1. Cell cycle analysis indicated that CHO-M1 cells had a similar cell cycle profile in the exponential growth phase, but cells rapidly accumulated in G1 phase just before confluence and did not progress through the cell cycle. This suggested that until confluence, proliferation of CHO-M1 was similar to parental CHO, but after confluence, it was inhibited and under G1 arrest. The specific antibody production rate of CHO-M1 was kept high, even after confluence, while that of parental CHO was drastically decreased in stationary phase. These results suggest that the desired cell line was successfully established and that high-energy beam irradiation could be an efficient mutagenic technique for breeding industrial cells.

  1. Agronomic Properties Assessed for Population of Recombinant Inbred Lines of Rye (Seceale cereale L. with Known Responses to Nutrient Deficiency Stress at the Seedling Stage

    Directory of Open Access Journals (Sweden)

    Milosz SMOLIK

    2013-05-01

    Full Text Available The experiment was established and carried out in light and slightly acidic soil in the years 2008-2010. The research material included one hundred thirty-eight RILs (F8:10 of rye with known response to nutrient stress were determined at the seedling stage obtained in vitro in mature embryos cultures and in two parental inbred lines and F1. The following factors were determined in the full maturity stage of each RIL: plant height, spike length, plant tillering, spikelet number per spike, grain number per spike, spike fertility, grain weight per spike, grain weight per plant and 1000 grain weight. Differences in the examined traits were found between parental inbred lines, and the heterosis effect was observed in F1 hybrid. The range of variability of the examined functional traits demonstrated in the population of RILs proved the transgression effect. The analyzed RILs were arranged into six groups and described with the use of Ward’s agglomerative method and grouping variables: spike fertility, grain weight per spike and per plant, thousand grain weight. Significant differences were found among other things between spike morphology and fertility traits and yield per a plant. The possibility of selection of RILs with extreme traits described both in the field and laboratory experiments in comparison with known tolerance to nutrient stress was described. The obtained correlation coefficients for the examined functional traits of RILs were in accordance with those published in reference literature. The correlation coefficients determined for selected seedling traits from the laboratory experiment and for selected traits of a mature plant may prove the usefulness of the test in studies on selection of genotypes desired for sustainable agriculture or for research on heritability of traits of tolerance to nutrient stress.

  2. Identification of candidate genes associated with cell wall digestibility and eQTL (expression quantitative trait loci) analysis in a Flint x Flint maize recombinant inbred line population.

    Science.gov (United States)

    Shi, Chun; Uzarowska, Anna; Ouzunova, Milena; Landbeck, Matthias; Wenzel, Gerhard; Lübberstedt, Thomas

    2007-01-18

    Cell-wall digestibility is the major target for improving the feeding value of forage maize. An understanding of the molecular basis for cell-wall digestibility is crucial towards breeding of highly digestible maize. 865 candidate ESTs for cell-wall digestibility were selected according to the analysis of expression profiles in 1) three sets of brown-midrib isogenic lines in the genetic background of inbreds 1332 (1332 and 1332 bm3), 5361 (5361 and 5361 bm3), and F2 (F2, F2 bm1, F2 bm2, and F2 bm3), 2) the contrasting extreme lines of FD (Flint x Dent, AS08 x AS 06), DD1 (Dent x Dent, AS11 x AS09), and DD2 (Dent x Dent, AS29 x AS30) mapping populations, and 3) two contrasting isogenic inbreds, AS20 and AS21. Out of those, 439 ESTs were assembled on our "Forage Quality Array", a small microarray specific for cell wall digestibility related experiments. Transcript profiles of 40 lines of a Flint x Flint population were monitored using the Forage Quality Array, which were contrasting for cell wall digestibility. Using t-tests (p quality groups. Using interval mapping, eQTL (LOD > or = 2.4) were detected for 20% (89 of 439) of the spotted ESTs. On average, these eQTL explained 39% of the transcription variation of the corresponding ESTs. Only 26% (23 of 89) ESTs detected a single eQTL. eQTL hotspots, containing greater than 5% of the total number of eQTL, were located in chromosomal bins 1.07, 1.12, 3.05, 8.03, and 9.04, respectively. Bin 3.05 was co-localized with a cell-wall digestibility related QTL cluster. 102 candidate genes for cell-wall digestibility were validated by genetical genomics approach. Although the cDNA array highlights gene types (the tested gene and any close family members), trans-acting factors or metabolic bottlenecks seem to play the major role in controlling heritable variation of gene expression related to cell-wall digestibility, since no in silico mapped ESTs were in the same location as their own eQTL. Transcriptional variation was

  3. Transfer of auxinic herbicide resistance from Brassica kaber to Brassica juncea and Brassica rapa through embryo rescue.

    Science.gov (United States)

    Mithila, J; Hall, J Christopher

    2013-01-01

    Auxinic herbicides are widely used in agriculture to selectively control broadleaf weeds. Prolonged use of auxinic herbicides has resulted in the evolution of resistance to these herbicides in some biotypes of Brassica kaber (wild mustard), a common weed in agricultural crops. In this study, auxinic herbicide resistance from B. kaber was transferred to Brassica juncea and Brassica rapa, two commercially important Brassica crops, by traditional breeding coupled with in vitro embryo rescue. A high frequency of embryo regeneration and hybrid plant establishment was achieved. Transfer of auxinic herbicide resistance from B. kaber to the hybrids was assessed by whole-plant screening of hybrids with dicamba, a widely used auxinic herbicide. Furthermore, the hybrids were tested for fertility (both pollen and pistil) and their ability to produce backcross progeny. The auxinic herbicide-resistant trait was introgressed into B. juncea by backcross breeding. DNA ploidy of the hybrids as well as of the backcross progeny was estimated by flow cytometry. Creation of auxinic herbicide-resistant Brassica crops by non-transgenic approaches should facilitate effective weed control, encourage less tillage, provide herbicide rotation options, minimize occurrence of herbicide resistance, and increase acceptance of these crops.

  4. Transcriptomics profiling of Indian mustard (Brassica juncea) under arsenate stress identifies key candidate genes and regulatory pathways.

    Science.gov (United States)

    Srivastava, Sudhakar; Srivastava, Ashish K; Sablok, Gaurav; Deshpande, Tejaswini U; Suprasanna, Penna

    2015-01-01

    Arsenic (As) is a non-essential element, a groundwater pollutant, whose uptake by plants produces toxic effects. The use of As-contaminated groundwater for irrigation can affect the crop productivity. Realizing the importance of the Brassica juncea as a crop plant in terms of oil-yield, there is a need to unravel mechanistic details of response to As stress and identify key functional genes and pathways. In this research, we studied time-dependent (4-96 h) transcriptome changes in roots and shoots of B. juncea under arsenate [As(V)] stress using Agilent platform. Among the whole transcriptome profiled genes, a total of 1,285 genes showed significant change in expression pattern upon As(V) exposure. The differentially expressed genes were categorized to various signaling pathways including hormones (jasmonate, abscisic acid, auxin, and ethylene) and kinases. Significant effects were also noticed on genes related to sulfur, nitrogen, CHO, and lipid metabolisms along with photosynthesis. Biochemical assays were conducted using specific inhibitors of glutathione and jasmonate biosynthesis, and kinases. The inhibitor studies revealed interconnection among sulfur metabolism, jasmonate, and kinase signaling pathways. In addition, various transposons also constituted a part of the altered transcriptome. Lastly, we profiled a set of key functional up- and down-regulated genes using real-time RT-PCR, which could act as an early indicators of the As stress.

  5. Exposure of Brassica juncea (L) to arsenic species in hydroponic medium: comparative analysis in accumulation and biochemical and transcriptional alterations.

    Science.gov (United States)

    Ahmad, Mohd Anwar; Gupta, Meetu

    2013-11-01

    Arsenic (As) contamination in the environment has attracted considerable attention worldwide. The objective of the present study was to see the comparative effect of As species As(III) and As(V) on accumulation, biochemical responses, and gene expression analysis in Brassica juncea var. Pusa Jaganath (PJn). Hydroponically grown 14-day-old seedlings of B. juncea were treated with different concentrations of As(III) and As(V). Accumulation of total As increased with increasing concentration of both As species and exposure time, mainly in roots. Reduction in seed germination, root-shoot length, chlorophyll, and protein content were observed with increasing concentration and exposure time of both As species, being more in As(III)-treated leaves. PJn variety showed that antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) and stress-related parameters (cysteine, proline, and malondialdehyde (MDA)) were stimulated and allows plant to tolerate both As species. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis in leaves showed significant changes in protein profile with more stringent effect with As(III) stress. Semiquantitative RT-PCR analysis showed regulation in expression of phytochelatin synthase (PCS), metallothionine-2 (MT-2), glutathione reductase (GR), and glutathione synthetase (GS) genes under both As(III) and As(V) stresses. Results suggested that accumulation and inhibition on physiological parameters differ according to the As species, while molecular and biochemical parameters showed a combinatorial type of tolerance mechanism against As(III) and As(V) stresses.

  6. Combined application of Triton X-100 and Sinorhizobium sp. Pb002 inoculum for the improvement of lead phytoextraction by Brassica juncea in EDTA amended soil.

    Science.gov (United States)

    Di Gregorio, Simona; Barbafieri, Meri; Lampis, Silvia; Sanangelantoni, Anna Maria; Tassi, Eliana; Vallini, Giovanni

    2006-04-01

    The process of EDTA-assisted lead phytoextraction from the Bovisa (Milan, Italy) brownfield soil was optimized in microcosms vegetated with Brassica juncea. An autochthonous plant growth-promoting rhizobacterium (PGPR), Sinorhizobium sp. Pb002, was isolated from the rhizosphere of B. juncea grown on the Pb-contaminated soil in presence of 2 mM EDTA. The strain was augmented (10(8) CFU g(-1) soil) in vegetated microcosms to stimulate B. juncea biomass production and, hence, its phytoextraction potential. Triton X-100 was also added to microcosms at 5 and 10 times the critical micelle concentration (cmc) to increase the permeability of root barriers to the EDTA-Pb complexes. Triton X-100 amendment determined an increase in Pb concentration within plant tissues. However it contextually exerted a phytotoxic effect. Sinorhizobium sp. Pb002 augmentation was crucial to plant survival in presence of both bioavailable lead and Triton X-100. The combination of the two treatments produced up to 56% increase in the efficiency of lead phytoextraction by B. juncea. The effects of these treatments on the structure of the soil bacterial community were evaluated by 16S rDNA denaturing gradient gel electrophoresis (DGGE).

  7. Path and ridge regression analysis of seed yield and seed yield components of Russian wildrye (Psathyrostachys juncea Nevski) under field conditions

    DEFF Research Database (Denmark)

    Wang, Quanzhen; Zhang, Tiejun; Cui, Jian

    2011-01-01

    The correlations among seed yield components, and their direct and indirect effects on the seed yield (Z) of Russina wildrye (Psathyrostachys juncea Nevski) were investigated. The seed yield components: fertile tillers m-2 (Y1), spikelets per fertile tillers (Y2), florets per spikelet- (Y3), seed...

  8. Glucose, stem dry weight variation, principal component and cluster analysis for some agronomic traits among 16 regenerated Crotalaria juncea accessions for potential cellulosic ethanol

    Science.gov (United States)

    The United State Department of Agriculture (USDA), Agricultural Research Service, (ARS), Plant Genetic Resources Conservation Unit’s (PGRCU) sunn hemp (Crotalaria juncea L.) germlasm collection consists of 22 accessions. Sixteen (16) accessions of the most seed productive were selected. These access...

  9. Measurement of the 1s2l3l’ Dielectronic Recombination Emission Line in Li-Like Ar and Its Contribution to the Faint X-Ray Feature Found in the Stacked Spectrum of Galaxy Clusters

    Science.gov (United States)

    Gall, Amy Christina; Silwal, Roshani; Dreiling, Joan; Borovik, Alexander; Ajello, Marco; Gillaspy, John; Kilgore, Ethan; Ralchenko, Yuri; Takacs, Endre

    2016-06-01

    Driven by the recent detection of an unidentified emission line previously reported at 3.55-3.57 keV in a stacked spectrum of galaxy clusters, we investigated the resonant DR process in Li-like Ar as a possible source of, or contributor to, the emission line. The Li-like transition 1s22l-1s2l3l’ was suggested to produce a 3.62 keV photon [1] near the unidentified line at 3.57 keV and was the primary focus of our investigation. Apart from the mentioned transitions, we have found other features that can be possible contributors to the emission in this region. The Electron Beam Ion Trap at NIST was used to produce and trap the highly-charged ions of argon. The energy of the quasi-monoenergetic electron beam was incremented in steps of 15 eV to scan over all of the Li-like Ar DR resonances. A Johann-type crystal spectrometer and a solid-state germanium detector were used to take x-ray measurements perpendicular to the electron beam. The DR cross sections were measured and normalized to the well-known photoionization cross sections using radiative recombination peaks in the measured spectra. Corrections for different instrument and method related effects such as charge state balance, electron beam space charge, and charge exchange have been considered. Our high-resolution crystal spectra allowed the experimental separation of features that are less than 2 eV apart. We have used a collisional radiative model NOMAD [2] aided by atomic data calculations by FAC [3] to interpret our observations and account for the corrections and uncertainties. Experimental results were compared to the AtomDB theoretical emission lines used to fit the galaxy cluster spectra containing the unidentified 3.57 keV line. These data points can be added benchmarks in the database and used to accurately interpret spectra from current x-ray satellites, including Hitomi, Chandra, and XMM-Newton x-ray observatories.[1] Bulbul E. et al., 2014, ApJ, 789, 13[2] Ralchenko Yu. et al., 2014, JQSRT, 71

  10. Uptake and speciation of vanadium in the rhizosphere soils of rape (Brassica juncea L.).

    Science.gov (United States)

    Tian, Li-Yan; Yang, Jin-Yan; Huang, Jen-How

    2015-06-01

    The response of rape (Brassica juncea L.) to different vanadium (V) speciation in rhizosphere soils was investigated in pot experiments using an agricultural soil containing 147 mg V kg(-1) supplemented with 0-500 mg V kg(-1) of pentavalent V [V(V)] and a mining soil containing 774 mg V kg(-1). Tetravalent V [V(IV)] accounted for 76.1 and 85.9 % of total V in the untreated agricultural soil and mining soil, respectively. The proportion of both V(V) and water-extractable V increased with increasing concentrations of V(V) in the agricultural soil. The growth of rape substantially reduced the concentrations of V(V) but not V(IV) in the rhizosphere soil, suggesting that V(V) was actively involved in the soil-rape interaction of V. Both soil V(V) and water-extractable V were negatively related to the total rape biomass, but were positively correlated with the concentration of root V. No such relationships were found for total V and soil V(IV). Together, these results indicate that soil V(V) and water-extractable V might better reflect the toxicity of V in soils than total V and soil V(IV). Rape accumulated V in the sequence: roots > > stem > leaf > seed. As indicated by the remarkably low root bioconcentration factor of V(V) (0.41-7.24 %), rape had a lower ability to accumulate V than other plants reported in the literature (14.6-298 %). Only a small fraction of V in rape roots was translocated to the aboveground organs (the translocation factor was 3.57-46.9 %). No V was detectable in seeds in the soils at 147 and 197 mg V kg(-1), and no seed was produced in the soils at higher V concentrations. Thus, the risk of V intake by humans via the consumption of rapeseed-based foods under normal conditions is considered to be lower than that of other plants.

  11. Effects of Nitrogen and Desferal Treatments on CROTALARIA's (Crotalaria juncea Roth) Biomass Production

    Science.gov (United States)

    László Phd, M., ,, Dr.

    2009-04-01

    Presently sustainable agriculture is vital to achieving food security poverty alleviation and environmental protection because land degradation and desertification has occurred in all the world over cutting across a broad spectrum of contrasts in climate, ecosystem types, land uses and socio/economic settings. For this reason improving integrated soil fertility management is appreciationed and has become a major issue of concern on the development plant nutrition and plant production agendas. On plant nutrition level mineral macronutrients so nitrogen and chelating agents of different microelements so Desferal- deferoxamin-methansulfonic are essential for plant growth and development. Crotalari juncea L. is a well-known nutrient indicator fodder and green manure crop with a high yield potential. Field experiment was carried out on a chernozem meadow soil (Kunság- region of Hungary, Kunmadaras) in partly of experiment series (6 years) in 2001. The ploughed layer of region soils contained with about 2.6-3.4% humus and 40-42% clay, had a humus stability index of 0.9-2.5 by Márton (1997), pH (H2O) of 6.5-7.7, pH (KCl) of 5.3-6.8, y1 of 6.7-13.3. The topsoil was poorly supplied with all five macronutrients (N-NO3 1 mg 100 g-1, AL-soluble P2O5 14 mg 100 g-1, AL-K2O 36 mg 100 g-1, Ca 330 mg 100 g-1, Mg 43 mg 100 g-1) and with all four micronutrients (0.5m HNO3 soluble Cu 1 mg kg-1, Zn 1 mg kg-1, Mn 9 mg kg-1, Fe 80 mg kg-1) according to soil analysis. The groundwater depth was 2-3 m. Nitrogen x Desferal (Novartis Pharma AG Basie, Switzerland, Suiza 500mg) x Genotype (Brazíl-EMBRAPA/CNPH, Brazilia-DF, India-University of Agricultural Sciences, Bangalore) x Time experiment involved 4Nx2Dx2Gx3T=48 treatments in 3 replications giving a total of 144 plots. The N levels were 0, 100, 200 and 300 kg ha-1 year-1, and desferal levels 0 and 20 kg ha-1 year-1 with a 100 kg ha-1 year-1 P2O5 and 120 kg ha-1 year-1 K2O basic fertilisation. The plot size had an area of 4x2=8 m2 with

  12. 利用重组自交系群体进行抗玉米矮花叶病研究%Research on Resistance to Maize Dwarf Mosaic Disease Using Recombinant Inbred Line in Maize

    Institute of Scientific and Technical Information of China (English)

    刘小红; 张红梅; 谭振波

    2006-01-01

    玉米矮花叶病在世界范围内广泛分布,且危害十分严重.因此正确认识玉米(Zea mays L.)对矮花叶病的抗性机制非常必要.基于此,本研究利用遗传差异较大的Mo17(高感玉米矮花叶病)和黄早四(高抗玉米矮花叶病)为亲本采用一粒传法构建了F9代重组自交系(Recombinant inbred line,RIL)分离群体,共256系.通过人工接甘蔗花叶病毒MDB株系(Sugarcane mosaic virus strain MDB,SCMV-MDB)对该分离群体进行了抗病性分析,统计了发病率和病情指数两项指标,根据这两项指标的频数分布图可知:玉米对矮花叶病的抗性主要由主基因控制,但不能排除还有微效基因的可能性.

  13. Study on the correlation between copper oxide nanoparticles induced growth suppression and enhanced lignification in Indian mustard (Brassica juncea L.).

    Science.gov (United States)

    Nair, Prakash M Gopalakrishnan; Chung, Ill Min

    2015-03-01

    In this study, the morphological, physiological and molecular level effects of copper oxide nanoparticles (CuONPs) were studied in an economically important oil seed crop Brassica juncea L. The possible involvement of lignification on shoot-root growth retardation was also studied. The seedlings were exposed to 0, 20, 50, 100, 200, 400 and 500mg/L of CuONPs in semi-solid half strength Murashige and Skoog medium under controlled growth chamber conditions for 14 days. Exposure to CuONPs resulted in suppression of shoot-growth, reduction in total chlorophyll and carotenoids contents as well modification of root system architecture such as shortening of primary and lateral roots. Significant increases in hydrogen peroxide formation, peroxidase enzyme activity and lignification of shoots and roots were observed. The lipid peroxidation levels increased significantly in shoots and roots of B. juncea seedlings. Phloroglucinol-HCl staining revealed enhanced lignification of shoot and roots. Gene expression studies revealed significant activation of CuZn superoxide dismutase (CuZnSOD) in roots at all concentrations of CuONPs exposure. In shoots significant up-regulation of CuZnSOD gene was observed upon exposure to 100, 200 and 400 mg/L of CuONPs exposure. However no change in the expression levels of MnSOD gene was observed in both stem and roots. The expression of catalase (CAT) and ascorbate peroxidase (APX) genes were also not changed in shoots. However, significant inhibition of CAT and APX genes were observed in roots of B. juncea plants under exposure to 100, 200, 400 and 500 mg/L of CuONPs exposure. The SOD enzyme activity significantly increased in roots under exposure to 50-500 mg/L of CuONPs and in shoots as a result of exposure to 100-500 mg/L of CuONPs. The APX activity significantly decreased in roots upon exposure to 50-500 mg/L of CuONPs. In shoots, the APX activity significantly decreased upon exposure to 200-500 mg/L of CuONPs.

  14. Resultados experimentais com a cultura da Crotalaria juncea L. L., como planta produtora de celulose para papel Results from field trials with Crotalaria juncea L. as fiber plant for paper pulp

    Directory of Open Access Journals (Sweden)

    Júlio César Medina

    1961-01-01

    Full Text Available São apresentados e discutidos os resultados de vários experimentos de campo realizados na Estação Experimental de Tatuí, do Instituto Agronômico, com a Crotalaria juncea L. como planta produtora de celulose para papel. Êsses resultados permitem fazer recomendações úteis sôbre o melhoramento técnico da cultura e da produção, no concernente às questões de variedades, espaçamento, densidade de semeação, época de semeação e época de colheita dos caules.This paper reports the results obtained in sunn hemp (Crotalaria juncea L. field trials carried out at the Tatui Experiment Station, São Paulo State. The trials were designed to study the effect of varieties, spacing x rate of seeding, time of sowing and time of harvesting on sunn hemp fiber production as a raw material for paper pulp. From the results achieved in these trials, the following recommendations can be made for growing sunn hemp as a fiber plant for paper pulp in the State of São Paulo. VARIETY: to use the common variety that is normally sowed for green manuring. TIME OF SOWING: to sow the seeds during october. RATE OF SEED SOWING AND SPACING: to sow the seeds at the rate of 100 lbs/ acre in rows spaced 8 inches apart. TIME OF HARVESTING: to harvest the stalks for mechanical fiber extraction when the seed pods are in the mature stage.

  15. Impact of Indian Mustard (Brassica juncea and Flax (Linum usitatissimum Seed Meal Applications on Soil Carbon, Nitrogen, and Microbial Dynamics

    Directory of Open Access Journals (Sweden)

    Autumn S. Wang

    2012-01-01

    Full Text Available There is a critical need to investigate how land application of dedicated biofuel oilseed meals affects soil ecosystems. In this study, mustard (Brassica juncea and flax (Linum usitatissimum seed meals and sorghum-sudangrass (Sorghum bicolor were added to soil at levels of 0, 1, 2.5, and 5% (w/w. Both the type of amendment and application rate affected soil organic C, total C & N, and C & N mineralization. Mustard meal amendment initially inhibited C mineralization as compared to flax, but >50% of mustard and flax organic C was mineralized within 51 d. Nitrogen mineralization was similar for flax and mustard, except for the 2.5% rate for which a lower proportion of mustard N was converted to nitrate. The mustard meal greatly impacted microbial community composition, appearing to select for specific fungal populations. The potential varying impacts of different oilseed meals on soil ecosystems should be considered when developing recommendations for land application.

  16. "Erussica", the intergeneric fertile somatic hybrid developed through protoplast fusion between Eruca sativa Lam. and Brassica juncea (L.) Czern.

    Science.gov (United States)

    Sikdar, S R; Chatterjee, G; Das, S; Sen, S K

    1990-04-01

    Hypocotyl calli-derived protoplasts of two cultivars of Brassica juncea (2n=36), a major oil-seed crop, were fused with normal as well as γ-irradiated mesophyll protoplasts of Eruca sativa (2n=22). The irradiation of the Eruca fusion partner increased the plating efficiency as well as the morphogenic potentiality of the fusion products over the normal fusion. Fertile plants could be regenerated from such fusion products. Analysis of 63 out of 181 plants regenerated showed that, indeed, 11 somatic hybrids (2n=58) and 10 partial somatic hybrids (chromosome number ranged between 50 and 56) had been obtained. Pollen viability (0%-82.9%) and seed set (0%-50%) of the hybrids indicated them to be useful for future studies.

  17. Serpentine bacteria influence metal translocation and bioconcentration of Brassica juncea and Ricinus communis grown in multi-metal polluted soils

    Directory of Open Access Journals (Sweden)

    Ying eMa

    2015-01-01

    Full Text Available The aim of this study was to assess the effects of inoculation of rhizosphere or endophytic bacteria (Psychrobacter sp. SRS8 and Pseudomonas sp. A3R3, respectively isolated from a serpentine environment on the plant growth and the translocation and accumulation of Ni, Zn and Fe by Brassica juncea and Ricinus communis on a multi-metal polluted serpentine soil (SS. Field collected SS was diluted to 0, 25, 50 and 75% with pristine soil in order to obtain a range of heavy metal concentrations and used in microcosm experiments. Regardless of inoculation with bacteria, the biomass of both plant species decreased with increase of the proportion of SS. Inoculation of plants with bacteria significantly increased the plant biomass and the heavy metal accumulation compared with non-inoculated control in the presence of different proportion of SS, which was attributed to the production of plant growth promoting and/or metal mobilizing metabolites by bacteria. However, SRS8 showed a maximum increase in the biomass of the test plants grown even in the treatment of 75% SS. In turn, A3R3 showed maximum effects on the accumulation of heavy metals in both plants. Regardless of inoculation of bacteria and proportion of SS, both plant species exhibited low values of bioconcentration factor (<1 for Ni and Fe. The inoculation of both bacterial strains significantly increased the translocation factor (TF of Ni while decreasing the TF of Zn in both plant species. Besides this contrasting effect, the TFs of all metals were < 1, indicating that all studied bacteria-plant combinations are suitable for phytostabilization. This study demonstrates that the bacterial isolates A3R3 and SRS8 improved the growth of B. juncea and R. communis in SS soils and have a great potential to be used as inoculants in phytostabilization scenarios of multi-metal contaminated soils.

  18. Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR.

    Directory of Open Access Journals (Sweden)

    Ruby Chandna

    Full Text Available The real time quantitative reverse transcription PCR (qRT-PCR is becoming increasingly important to gain insight into function of genes. Given the increased sensitivity, ease and reproducibility of qRT-PCR, the requirement of suitable reference genes for normalization has become important and stringent. It is now known that the expression of internal control genes in living organism vary considerably during developmental stages and under different experimental conditions. For economically important Brassica crops, only a couple of reference genes are reported till date. In this study, expression stability of 12 candidate reference genes including ACT2, ELFA, GAPDH, TUA, UBQ9 (traditional housekeeping genes, ACP, CAC, SNF, TIPS-41, TMD, TSB and ZNF (new candidate reference genes, in a diverse set of 49 tissue samples representing different developmental stages, stress and hormone treated conditions and cultivars of Brassica juncea has been validated. For the normalization of vegetative stages the ELFA, ACT2, CAC and TIPS-41 combination would be appropriate whereas TIPS-41 along with CAC would be suitable for normalization of reproductive stages. A combination of GAPDH, TUA, TIPS-41 and CAC were identified as the most suitable reference genes for total developmental stages. In various stress and hormone treated samples, UBQ9 and TIPS-41 had the most stable expression. Across five cultivars of B. juncea, the expression of CAC and TIPS-41 did not vary significantly and were identified as the most stably expressed reference genes. This study provides comprehensive information that the new reference genes selected herein performed better than the traditional housekeeping genes. The selection of most suitable reference genes depends on the experimental conditions, and is tissue and cultivar-specific. Further, to attain accuracy in the results more than one reference genes are necessary for normalization.

  19. recombinant inbred lines intercropped with oat

    African Journals Online (AJOL)

    ADMIN

    was planted to winter wheat. The field was ... cropping, the interaction of RILs by intercropping methods and .... barley RILs for the characters evaluated. Environments. Character. Genotype. B. B01 ...... Thesis submitted to Department of Plant.

  20. Optical recombination lines and temperature fluctuations

    Directory of Open Access Journals (Sweden)

    X. W. Liu

    2002-01-01

    Full Text Available Hace tres d ecadas que Peimbert encontr o que las temperaturas del salto de Balmer en regiones H II y planetarias (PNe son sistem aticamente menores a las obtenidas con las l neas del [O III]. Sugiri o que debido a la existencia de uctuaciones de la temperatura, las abundancias de elementos pesados derivadas de l neas de excitaci on colisional (CELs eran sistem aticamente subestimadas. En este trabajo hago una revisi on de las determinaciones recientes de elementos pesados en PNe con l neas d ebiles de elementos pesados (ORLs. En todas las especies i onicas estudiadas, las ORLs dan abundancias mayores que las CELs. El cociente de abundancias ORL a CEL var a de objeto a objeto y est a correlacionado con la diferencia de temperaturas del [O III] y del salto de Balmer. Las uctuaciones de temperatura y densidad solas no pueden explicar las grandes discrepancias en abundancias obtenidas con ORL y CEL en un n umero peque~no de nebulosas. Se sugiere que estos casos extremos pueden contener regiones ricas en metales similares a las observadas en PN rejuvenecidas, como Abell 30.

  1. RECOMBINATION AND COLLISIONALLY EXCITED BALMER LINES

    Directory of Open Access Journals (Sweden)

    A. C. Raga

    2015-01-01

    Full Text Available Presentamos un modelo para el equilibrio estad ́ıstico de lo s niveles de H, considerando recombinaciones a los niveles excitados, exc itaciones colisionales par- tiendo desde el nivel fundamental y transiciones radiativa s espont ́aneas. Este pro- blema tiene una simple solucion en t ́erminos de la “matriz de cascada”, que des- cribe una cascada de transiciones espont ́aneas alimentada tanto por recombina- ciones como por excitaciones colisionales. Las prediccion es resultantes de cocientes de l ́ıneas de Balmer muestran una transici ́on entre un r ́egi men de baja temperatura y uno de alta temperatura (dominados por recombinaciones y p or excitaciones co- lisionales, respectivamente, ambos con s ́olo una depende ncia d ́ebil de la tempe- ratura. Esta clara caracter ́ıstica permite una diferencia ci ́on observacional directa entre regiones de l ́ıneas de Balmer de recombinaci ́on y regi ones con l ́ıneas excitadas colisionalmente. Encontramos que para un gas en equilibrio coronal las l ́ıneas de H α y H β se excitan colisionalmente a todas las temperaturas. Para o btener l ́ıneas de H α y H β de recombinaci ́on, es necesario tener fracciones de ioniza ci ́on de H sub- tancialmente mayores que la de equilibrio coronal (por ejem plo, como las presentes en un gas fotoionizado.

  2. Recombinant Technology and Probiotics

    Directory of Open Access Journals (Sweden)

    Icy D’Silva

    2011-09-01

    Full Text Available Recombinant technology has led the way to monumental advances in the development of useful molecules, including the development of safe probiotics. The development of novel approaches using recombinant technology and probiotics that allow accurate targeting of therapeutics to the mucosa is an interesting area of research. The creation and use of recombinant probiotics expressing recombinantovalbumin, recombinant ovalbumin mutants and yet-to-be-designed recombinant hypo/non-allergenic molecules offer the opportunity to further investigate their effects for food, nutrition, environment andhealth. This review highlights advances in native probiotics and recombinant probiotics expressing native and recombinant molecules for food, nutrition, environment and health.

  3. Recombinant adeno-associated virus-mediated global anterograde delivery of glial cell line-derived neurotrophic factor to the spinal cord: comparison of rubrospinal and corticospinal tracts in the rat.

    Science.gov (United States)

    Foust, Kevin D; Flotte, Terence R; Reier, Paul J; Mandel, Ronald J

    2008-01-01

    Amyotrophic lateral sclerosis (ALS) is characterized by progressive loss of spinal lower motoneurons. Gene delivery is a promising strategy to deliver therapeutic molecules to these vulnerable cells. However, definition of an optimal route of delivery capable of accessing neurons over a considerable extent of the neuraxis represents a significant logistical problem. Intramuscular vector injections are not ideal as this approach would involve hundreds of injections to completely treat an ALS patient and also would be dependent on retrograde transport of the viral platform of choice. Alternatively, upper motoneurons could deliver trophic factors over considerable distances by anterograde transport after a relatively localized intracerebral injection. To test this approach, the present study was designed to compare the corticospinal (CST) and rubrospinal (RST) tracts for their ability to transport recombinant adeno-associated virus serotype 5 (rAAV5)-derived green fluorescent protein (GFP) or glial cell line-derived neurotrophic factor (GDNF) to the spinal cord. Unilateral injections of rAAV5-GFP into the red nucleus (RN) or motor cortex of normal rats produced GFP-positive fibers in the appropriate descending tracts extending to the lumbar spinal cord. For both tracts, GFP-positive axonal projections into the spinal gray matter were consistently observed. GDNF immunohistochemistry demonstrated that confirmed RN injections resulted in GDNF-positive fibers projecting into spinal gray matter as seen in the GFP group. In contrast, confirmed cortical rAAV5-GDNF injections resulted in less evident staining in spinal cord. Spinal cord GDNF levels were elevated at distances up to 72 mm from the injection sites, and confirmed that RST-related GDNF transport to spinal cord surpassed CST-associated delivery.

  4. Recombinant Technology and Probiotics

    OpenAIRE

    Icy D’Silva

    2011-01-01

    Recombinant technology has led the way to monumental advances in the development of useful molecules, including the development of safe probiotics. The development of novel approaches using recombinant technology and probiotics that allow accurate targeting of therapeutics to the mucosa is an interesting area of research. The creation and use of recombinant probiotics expressing recombinantovalbumin, recombinant ovalbumin mutants and yet-to-be-designed recombinant hypo/non-allergenic molecule...

  5. QTL Analysis of Kernel Characteristics Using a Recombinant Inbred Lines (RILs) Population Derived from the Cross of Tiriticum polonicum L. and Triticum aestivum L. Line “Zhong 13”%波兰小麦×普通小麦品系“中13”RIL群体籽粒特性的QTL定位

    Institute of Scientific and Technical Information of China (English)

    李美霞; 杨睿; 李有梅; 崔桂宾; 王竹林; 奚亚军; 刘曙东

    2012-01-01

    小麦籽粒特性与籽粒产量和品质密切相关。本研究以波兰小麦(Tiriticum polonicum L.)×普通小麦(Triticum aestivum L.)品系“中13”杂交组合衍生的99个F8重组自交系(Recombinant inbred lines, RIL)群体为材料,利用SSR分子标记构建连锁遗传图谱。根据两年实验数据,利用复合区间作图法对粒重、粒长和粒宽3个籽粒特性相关性状进行了QTL定位分析,共检测到12个与籽粒特性相关的加性QTL位点。其中,3个粒重QTL,1个位于1A染色体上,另外2个都在2A染色体上,单个QTL可解释表型变异的13.35%~20.04%;5个粒长QTL,其中2个位于2A染色体上,其余3个分别位于3A、5A和2B染色体上,单个QTL可解释表型变异的 8.53%~21.03%;4个粒宽QTL,分别位于1A、2A、3B和 5B染色体上,单个QTL可解释表型变异的9.76%~40.79%。在2A染色体上共检测到5个籽粒特性相关性状的QTL,表明2A染色体与籽粒特性关系密切。%Wheat kernel characteristics are associated with kernel yield and quality. In this study, the QTLs of grain weight, grain length and grain width were analyzed using SSR markers in a population which consisted of 99 F8 recombinant inbred lines (RILs) derived from the cross of Poland wheat (Triticum polonicum L.) and Common Wheat (Triticum aestivum L.) Zhong 13. The ANOVA showed that there was highly significant difference between Poland Wheat and Zhong 13 in all three traits in 2010 and 2011. It was clear that the hereditary basis between two parents on grain weight, grain length and grain width was very different. A significant difference among recombinant inbred lines (RIL) was found in grain weight in two years, a highly significant difference among recombinant inbred lines (RIL) was found in grain length and grain width in two years. Of 355 pairs of polymorphic SSR primers between parents, there were 173 pairs of SSR primers with polymorphism among recombinant

  6. 新麦草抗除草剂自然变异体的筛选%Spontancous Mutation Resistant to Herbicide of Psathyrostachys juncea(Fisch.)Nevski.

    Institute of Scientific and Technical Information of China (English)

    王宝地

    2011-01-01

    在细胞和植株两个水平上,筛选野生新麦草抗除草剂草甘膦自然变异体.在适宜的选择压力下,野生新麦草在两个水平上均有一定频率的抗除草剂自然变异体或突变型产生.100 mg/L的草甘膦是野生新麦草的抗性愈伤组织适宜的筛选浓度,抗性愈伤组织出现的频率为0.23%.用高于大田推荐剂量的草甘膦喷施野生新麦草植株,结果出现2×10-6的抗性个体.%Spontaneous mutants resistant to herbicide were screened on cell and plant levels of wild Psathy rostachys juncea (Fisch.) Nevski. On proper selection pressure, spontaneous mutants resistant to herbicide on both levels of wild Psath yrostachys juncea (Fisch.) Nevski. were existed. At the level of 100 mg/L glyphosate, the frequency of resistant calli in Psathy rostachys juncea (Fisch.) Nevski. was 0.23% and 100 mg/L was the comfortable screening dosage. 2×l0-6 mutants resistant to herbicide were obtained when Psathy rostachys juncea (Fisch.) Nevski. was sprayed with higher than recommended maximum dosages of herbicides in field.

  7. Growth, tolerance efficiency and phytoremediation potential of Ricinus communis (L.) and Brassica juncea (L.) in salinity and drought affected cadmium contaminated soil.

    Science.gov (United States)

    Bauddh, Kuldeep; Singh, Rana P

    2012-11-01

    We have previously reported that Ricinus communis (castor) is more tolerant to soil cadmium (Cd) and more efficient for Cd phytoremediation than Brassica juncea (Indian mustard) (Bauddh and Singh, 2012). In the present study, R. communis was found more tolerant to salinity and drought in presence of Cd and removed more Cd in a given time than Indian mustard. R. communis produced 23 and twelve folds higher biomass in terms of fresh weight and dry weight, respectively than that in B. juncea during three months when grown in Cd contaminated soil in presence of 100mM NaCl salinity and ten day water withdrawal based drought at 90 day after sowing (DAS). Castor plants showed stronger self-protection ability in form of proline bioaccumulation (r(2)=0.949) than Indian mustard (r(2)=0.932), whereas a lower r(2) for malondialdehyde (MDA) and total soluble protein in R. communis (r(2)=0.914 and r(2)=0.915, respectively) than that of B. juncea (r(2)=0.947 and r(2)=0.927, respectively) indicated a greater damage to cell membrane in Indian mustard during the multiple stress conditions. Though, the amount of Cd accumulated in the roots and shoots of Indian mustard was higher as per unit biomass than that in castor, total removal of the metal from soil was much higher in castor on per plant basis in the same period in presence of the stresses. R. communis accumulated about seventeen and 1.5 fold higher Cd in their roots and shoots, respectively than that of B. juncea in 90 DAS under the multiple stresses. Salinity alone enhanced Cd uptake, whereas drought stress reduced its uptake in both the plants. Copyright © 2012 Elsevier Inc. All rights reserved.

  8. Isolation of phytase-producing bacteria from Himalayan soils and their effect on growth and phosphorus uptake of Indian mustard (Brassica juncea).

    Science.gov (United States)

    Kumar, Vinod; Singh, Prashant; Jorquera, Milko A; Sangwan, Punesh; Kumar, Piyush; Verma, A K; Agrawal, Sanjeev

    2013-08-01

    Phytase-producing bacteria (PPB) is being investigated as plant growth promoting rhizobacteria (PGPR) to improve the phosphorus (P) nutrition and growth of plants grown in soil with high phytate content. Phytate is dominant organic P forms in many soils and must be hydrolyzed to be available for plants. Indian mustard (Brassica juncea) is a plant with economic importance in agriculture and phytoremediation, therefore biotechnological tools to improve growth and environmental stress tolerance are needed. In this study, we isolated and characterized PPB from Himalayan soils and evaluated their effect on growth and P uptake by B. juncea under greenhouse conditions. Sixty five PPB were isolated and based on phytate hydrolysis, three efficient PPB were chosen and identified as Acromobacter sp. PB-01, Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13. Selected PPB showed ability to grow at wide range of pH, temperature and salt concentrations as well as to harbour diverse PGPR activities, such as: solubilization of insoluble Ca-phosphate (193-642 μg ml(-1)), production of phytohormone indole acetic acid (5-39 μg ml(-1)) and siderophore. Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 showed 50 and 70 % inhibition of phytopathogen Rhizoctonia solani, respectively. Greenhouse potting assay also showed that the bacterization of B. juncea seeds with Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 significantly increased the biomass and P content in 30 days old seedlings. This study reveals the potential of PPB as PGPR to improve the growth of B. juncea.

  9. Uptake of heavy metals and As by Brassica juncea grown in a contaminated soil in Aznalcollar (Spain): The effect of soil amendments

    Energy Technology Data Exchange (ETDEWEB)

    Clemente, Rafael [Department of Soil and Water Conservation and Organic Waste Management. Centro de Edafologia y Biologia Aplicada del Segura, CSIC. Apartado 164, 30100 Espinardo, Murcia (Spain); Walker, David J. [Department of Soil and Water Conservation and Organic Waste Management. Centro de Edafologia y Biologia Aplicada del Segura, CSIC. Apartado 164, 30100 Espinardo, Murcia (Spain); Bernal, M. Pilar [Department of Soil and Water Conservation and Organic Waste Management. Centro de Edafologia y Biologia Aplicada del Segura, CSIC. Apartado 164, 30100 Espinardo, Murcia (Spain)]. E-mail: pbernal@cebas.csic.es

    2005-11-15

    Two crops of Brassica juncea (L.) Czern. were grown in a field experiment, at the site affected by the toxic spillage of acidic, metal-rich waste in Aznalcollar (Seville, Spain), to study its metal accumulation and the feasibility of its use for metal phytoextraction. The effects of organic soil amendments (cow manure and mature compost) and lime on biomass production and plant survival were also assessed; plots without organic amendment and without lime were used as controls. Plots, with or without organic amendment, having pH<5 were limed for the second crop. Soil acidification conditioned plant growth and metal accumulation. The addition of lime and the organic amendments achieved higher plant biomass production, although effects concerning metal bioavailability and accumulation were masked somewhat by pH variability with time and between and within plots. Tissue metal concentrations of B. juncea were elevated for Zn, Cu and Pb, especially in leaves of plants from plots with low pH values (maxima of 2029, 71 and 55 {mu}g g{sup -1}, respectively). The total uptake of heavy metals in the plants was relatively low, emphasising the problems faced when attempting to employ phytoextraction for clean-up of pluri-contaminated sites. - Although organic amendments improved soil conditions and plant growth, the phytoextraction capacity of Brassica juncea (cv. Z1) is too low for efficient soil remediation.

  10. Changes in C-N metabolism under elevated CO2 and temperature in Indian mustard (Brassica juncea L.): an adaptation strategy under climate change scenario.

    Science.gov (United States)

    Seth, Chandra Shekhar; Misra, Virendra

    2014-11-01

    The present study was performed to investigate the possible role of carbon (C) and nitrogen (N) metabolism in adaptation of Indian mustard (Brassica juncea L.) growing under ambient (370 ± 15 ppm) and elevated CO2 (700 ± 15 ppm), and jointly in elevated CO2 and temperature (30/22 °C for day/night). The key enzymes responsible for C-N metabolism were studied in different samples of Brassica juncea L. collected from ambient (AMB), elevated (ELE) and ELExT growth conditions. Total percent amount of C and N in leaves were particularly estimated to establish a clear understanding of aforesaid metabolism in plant adaptation. Furthermore, key morphological and physiological parameters such as plant height, leaf area index, dry biomass, net photosynthetic rate, stomatal conductance, transpiration, total protein and chlorophyll contents were also studied in relation to C/N metabolism. The results indicated that the C-metabolizing enzymes, such as (ribulose-1,5-bisphosphate carboxylase/oxygenase, phosphoenolpyruvate carboxylase, malate dehydrogenase, NAD-malic enzyme, NADP-malic enzyme and citrate synthase) and the N-metabolizing enzymes, such as (aspartate amino transferase, glutamine synthetase, nitrate reductase and nitrite reductase) showed significantly (P ELExT > AMB growth conditions. This is also evident by significant (P adaptation in Brassica juncea L. against elevated CO2 and temperature prevailing in climate change scenarios.

  11. Alleviation of selenium toxicity in Brassica juncea L.: salicylic acid-mediated modulation in toxicity indicators, stress modulators, and sulfur-related gene transcripts.

    Science.gov (United States)

    Gupta, Shikha; Gupta, Meetu

    2016-11-01

    The present work reveals the response of different doses of selenium (Se) and alleviating effect of salicylic acid (SA) on Se-stressed Brassica juncea seedlings. Selenium, a micronutrient, is essential for both humans and animals but is toxic at higher doses. Its beneficial role for the survival of plants, however, is still debatable. On the other hand, SA, a phenolic compound, is known to have specific responses under environmental stresses. Experiments were conducted using leaves of hydroponically grown seedlings of Pusa bold (PB) variety of B. juncea, treated with different concentrations of Se (50, 150, 300 μM) for 24- and 96-h exposure times. Increasing Se concentrations inhibited growth and, caused lipid peroxidation, concomitantly increased stress modulators (proline, cysteine, SOD, CAT) along with sulfur-related gene transcripts (LAST, APS, APR, GR, OASL, MT-2, PCS) in Brassica seedlings. On the basis of the above studied parameters, maximum inhibition in growth was observed at 300 μM Se after 96-h exposure time. Further, co-application of SA along with 300 μM Se helped to mitigate Se stress, as shown by improved levels of growth parameters, toxicity indicators (chlorophyll, protein, MDA), stress modulators (proline, cysteine, SOD, and CAT), and expression of sulfur-related genes as compared to Se-treated seedlings alone. Altogether, this study revealed that Se + SA combinations improved seedling morphology and were effective in alleviation of Se stress in PB variety of B. juncea.

  12. ABCC2基因过表达细胞株的建立及鉴定%Construction and identification of the recombinant cell line over-expressing ABCC2 gene

    Institute of Scientific and Technical Information of China (English)

    魏丹芸; 张洪; 彭锐; 张英

    2016-01-01

    Objective To construct the lentiviral vecter carrying human ABCC2 gene,then to screen out the stable cell line over -ex-pressing ABCC2 after package and transfection to HEK293 cells,thus to provide cell model to determine substrate drugs of multidrug re-sistance protein 2(MRP2)in vitro.Methods The primers were designed according to the cDNA sequence of ABCC2 gene from Gen-Bank.The gene were amplified by PCR and connected to lentiviral vector PEZ -LV105.The recombined lentiviral vector was pack-aged and transfected to HEK293 cells.After puromycin screening,HEK293 cells over -expressing human ABCC2 gene were selected and cloned.Finally,gene sequencing,real -time quantitative PCR(RTQ -PCR)and Western blot assays were performed for identifica-tion.Results RTQ -PCR showed that the ABCC2 mRNA in HEK293 cells transfected with exogenous ABCC2 gene was about 320 times higher than that of normal HEK293 cells and blank carrier HEK293 cells.MRP2 protein level in HEK293 cells with ABCC2 gene transfection was nearly 150 times higher than that of normal HEK293 cells and blank carrier HEK293 cells according to the Western blot.Conclusions Stable recombinant cell line over -expressing ABCC2 was successfully generated.The cell line could be useful in the confirmation of substrate drugs of multidrug resistance protein 2(MRP2)and the transport mechanism in vitro.%目的:构建携带 ABCC2基因的慢病毒载体,转染 HEK293细胞,筛选出稳定过表达 ABCC2基因的细胞株并进行鉴定,为体外实验确定与多药耐药相关蛋白2(MRP2)外排转运相关的底物药物及其转运机制提供细胞模型。方法根据 GenBank提供的 ABCC2基因 cDNA 序列设计引物,PCR 扩增该基因并将其连接至慢病毒载体 PZE -LV105,包装病毒并感染 HEK293细胞。用嘌呤霉素进行筛选得到过表达 ABCC2基因的稳转细胞株,通过基因测序、实时荧光定量 PCR(RTQ -PCR)和蛋白免疫印迹(Western blot)对稳转细

  13. Serumfree culture of the suspension cell line QB-Tn9-4s of the cabbage looper, Trichoplusia ni, is highly productive for virus replication and recombinant protein expression.

    Science.gov (United States)

    Zheng, Gui-Ling; Zhou, Hong-Xu; Li, Chang-You

    2014-02-12

    Serumfree cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serumfree Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 ×10(6) cells/mL in serumfree culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 10(6) cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 ×10(7) TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of β-galactosidase at six days postinfection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×10(4) IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serumfree culture.

  14. Serum-Free Culture of the Suspension Cell Line QB-Tn9-4s of the Cabbage Looper, Trichoplusia ni, is Highly Productive for Virus Replication and Recombinant Protein Expression

    Science.gov (United States)

    Zheng, Gui-Ling; Zhou, Hong-Xu; Li, Chang-You

    2014-01-01

    Serum-free cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serum-free Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 × 106 cells/mL in serum-free culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 106 cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 × 107 TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of β-galactosidase at six days post-infection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×104 IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serum-free culture. PMID:25373171

  15. Therapeutic Recombinant Monoclonal Antibodies

    Science.gov (United States)

    Bakhtiar, Ray

    2012-01-01

    During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

  16. Therapeutic Recombinant Monoclonal Antibodies

    Science.gov (United States)

    Bakhtiar, Ray

    2012-01-01

    During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

  17. Desempenho agronômico de linhas endogâmicas recombinadas de pimenta em dois sistemas de cultivo Agronomic performance of recombinant chili pepper inbred lines in two cultivation systems

    Directory of Open Access Journals (Sweden)

    Sarah Ola Moreira

    2009-08-01

    .Climate and management diversity play an important role in crop production, since these characteristics are related to genotype and cultivation environment. The aim of this study was to evaluate the effects of genotype, environment and genotype-environmental interaction (GE in 12 recombinant inbred lines of C. annuum. These lines were originated from crosses between the following accessions: a sweet pepper cultivar (susceptible to bacterial spot and a chili pepper (resistant to bacterial spot. Two tests were conducted, the first in a conventional agriculture system, under field conditions and the second in ecological or organic system, under greenhouse conditions. The field experiment was carried out in Campos dos Goytacazes, RJ. The greenhouse experiment was conducted in Seropédica, RJ. Both experiments were performed using a randomized block design with three replications. The measured variables were total fruit number (TFN; total fruit weight (TFW; mean fruit weight (MFW; fruit length (FL; fruit diameter (FD; length/diameter fruit ratio (FL/FD and capsaicin presence (CAPS. Data obtained for each experimental condition were submitted to variance analysis, joint variance analysis, and partitioning of genotype environment interaction. Genotype and GE quadratic components, genotypic determination coefficient, genetic variation coefficient and variation index were determined. The GE interaction was significant for TFN, TFW, MFN, FD, and FL/FD. Cultivation under greenhouse conditions was responsible for the highest mean performance for every trait studied but FL. Values observed for genetic variation were greater than values obtained for environmental variation. For field conditions, the high yield ability observed for line 2 (pungent is suggestive of a potential to be used as a commercial genotype. For greenhouse conditions, line 8 (pungent, had higher fruit number yield. Line 1 (non-pungent also seems promising for commercial cultivation under both conditions.

  18. Antagonistic experimental coevolution with a parasite increases host recombination frequency

    Directory of Open Access Journals (Sweden)

    Kerstes Niels AG

    2012-02-01

    Full Text Available Abstract Background One of the big remaining challenges in evolutionary biology is to understand the evolution and maintenance of meiotic recombination. As recombination breaks down successful genotypes, it should be selected for only under very limited conditions. Yet, recombination is very common and phylogenetically widespread. The Red Queen Hypothesis is one of the most prominent hypotheses for the adaptive value of recombination and sexual reproduction. The Red Queen Hypothesis predicts an advantage of recombination for hosts that are coevolving with their parasites. We tested predictions of the hypothesis with experimental coevolution using the red flour beetle, Tribolium castaneum, and its microsporidian parasite, Nosema whitei. Results By measuring recombination directly in the individuals under selection, we found that recombination in the host population was increased after 11 generations of coevolution. Detailed insights into genotypic and phenotypic changes occurring during the coevolution experiment furthermore helped us to reconstruct the coevolutionary dynamics that were associated with this increase in recombination frequency. As coevolved lines maintained higher genetic diversity than control lines, and because there was no evidence for heterozygote advantage or for a plastic response of recombination to infection, the observed increase in recombination most likely represented an adaptive host response under Red Queen dynamics. Conclusions This study provides direct, experimental evidence for an increase in recombination frequency under host-parasite coevolution in an obligatory outcrossing species. Combined with earlier results, the Red Queen process is the most likely explanation for this observation.

  19. Analysis of rice nitrogen use efficiency based on recombinant inbred line population%基于重组自交系群体水稻氮素利用效率分析和利用

    Institute of Scientific and Technical Information of China (English)

    阮新民; 施伏芝; 从夕汉; 罗志祥

    2016-01-01

    Nitrogen is the most critical input limiting rice productivity. Due to increasing fertilizer costs and environmental concerns, nitrogen use efficiency (NUE) is hotly debated in the scientific community. To explore the absorption, utilization and main traits distribution patter of nitrogen in recombinant inbred lines (RILs), a field experiment was conducted to evaluate potential NUE of rice (Oryza sativaL.). The split-plot experiment had two treatments (one without nitrogen fertilizer and the other with 150 kg·hm-2 of nitrogen) for populations of RILs, the parents and the check line Q149. The relationship between NUE and the main agronomic characteristics of RILs population was determined using statistical correlation, clustering, principal component analyses. The results showed that the variation coefficient of NUE was larger under 150 kg·hm-2 nitrogen treatment than under non-nitrogen treatment. Proper increase in nitrogen fertilization was beneficial to nitrogen content in rice panicle, stem and leaf, and to total dry matter weight of single plant. Significantly positive correlations were noted between dry matter production efficiency and plant high, panicle length, stem weight per plant, total dry matter weight of single plant under two nitrogen treatments. Also under both treatments, significantly negative correlations were noted between dry matter production efficiency and the contents of nitrogen in rice stem, leaf and panicle. There were positive correlations between grain production efficiency and grain weight per plant, seed setting rate, thousand-seed weight, total number of grains per panicle, and spike length. Equally, there were negative correlations between grain production efficiency and stem weight per plant, leaf nitrogen content, leaf weight per plant and total amount of nitrogen per plant. Stepwise regression analysis indicated that nitrogen content in stem and panicle, and stem weight per plant had significant effects on dry matter NUE

  20. Serpentine bacteria influence metal translocation and bioconcentration of Brassica juncea and Ricinus communis grown in multi-metal polluted soils.

    Science.gov (United States)

    Ma, Ying; Rajkumar, Mani; Rocha, Inês; Oliveira, Rui S; Freitas, Helena

    2014-01-01

    The aim of this study was to assess the effects of inoculation of rhizosphere or endophytic bacteria (Psychrobacter sp. SRS8 and Pseudomonas sp. A3R3, respectively) isolated from a serpentine environment on the plant growth and the translocation and accumulation of Ni, Zn, and Fe by Brassica juncea and Ricinus communis on a multi-metal polluted serpentine soil (SS). Field collected SS was diluted to 0, 25, 50, and 75% with pristine soil in order to obtain a range of heavy metal concentrations and used in microcosm experiments. Regardless of inoculation with bacteria, the biomass of both plant species decreased with increase of the proportion of SS. Inoculation of plants with bacteria significantly increased the plant biomass and the heavy metal accumulation compared with non-inoculated control in the presence of different proportion of SS, which was attributed to the production of plant growth promoting and/or metal mobilizing metabolites by bacteria. However, SRS8 showed a maximum increase in the biomass of the test plants grown even in the treatment of 75% SS. In turn, A3R3 showed maximum effects on the accumulation of heavy metals in both plants. Regardless of inoculation of bacteria and proportion of SS, both plant species exhibited low values of bioconcentration factor (communis in SS soils and have a great potential to be used as inoculants in phytostabilization scenarios of multi-metal contaminated soils.

  1. Effect of sunn hemp (Crotalaria juncea L.) cutting date and planting density on weed suppression in Georgia, USA.

    Science.gov (United States)

    Morris, J Bradley; Chase, Carlene; Treadwell, Danielle; Koenig, Rosie; Cho, Alyssa; Morales-Payan, Jose Pable; Murphy, Tim; Antonious, George F

    2015-01-01

    A field study was conducted in 2008 and 2009 at the USDA, ARS, Plant Genetic Resources Conservation Unit in Griffin, GA, to investigate weed suppression by sunn hemp (Crotalaria juncea L). The objectives were to (1) evaluate the effects of apical meristem removal (AMR) at three dates [5, 6, and 7 wks after planting (WAP) on May 14, 2008 and May 21, 2009] and (2) assess the impact of seeding rates (11, 28, and 45 kg ha(-1)) on weed biomass reduction. Weed species were identified at 4, 8, and 12 wks after sunn hemp planting. Sunn hemp cutting date had no significant effect on weed suppression in 2008 but significant differences for grass weeds at 4, 8, and 12 WAP and for yellow nutsedge at 8 and 12 WAP did occur when compared to the control in 2009. In comparison to the sunn hemp-free control plot in 2009, all three seeding rates had reduced grass weed dry weights at 4, 8, and 12 WAP. The total mass of yellow nutsedge when grown with sunn hemp was reduced compared to the total mass of yellow nutsedge grown in the weedy check for all seeding rates at 8 and 12 WAP. Lower grass weed biomass was observed by 12 WAP for cutting dates and seeding rates during 2008 and 2009. Sunn hemp cutting date and seeding rate reduced branch numbers in both years. The reduction in sunn hemp seeding rates revealed a decrease in weed populations.

  2. Pathogens present on vegetative organs and seeds of white mustard (Sinapis alba L. and chinese mustad (Brassica juncea L.

    Directory of Open Access Journals (Sweden)

    Barbara Majchrzak

    2013-12-01

    Full Text Available The research was conducted in the years 1999-2001. The aim of the research was to determine the health condition of overground parts and seeds of white niuslard (Sinapis alba L. cv. Metex and chinese mustard (Brassica juncea L. cv. Małopolska. In all the years of the research alternaria blight was found on the leaves of white mustard which injury index ranged from 5,6% in 2001 to 17,6% in 200O. The most dangerous disease of chinese mustard also was alternaria blight and its symptoms were found on leaves and siliques. The strongest infection of leaves was in 2000 (50% and the weakest in 2001 (6,7%. In all the years of the research siliques were rather weak infected (50-8,89%. Besides powdery mildew was found on chinese mustard which injury index ranged from 0,3% in 1999 to 32,3% in 2000. Intensity of diseases was affected generally by the weather conditions. From the seeds of white mustard and chinese mustard were isolated respectively 263 and 137 colonies. Alternaria alternata was the most numerous species which makes respectively 60,9% and 42,3% isolates. Among the fungi pathogenic for white and chinese mustard were also isolated: A. brassicae, Botrytis cinerea and Rhizoctonia solami.

  3. Heavy Metal Contents and Physical Parameters of Aegiceras corniculatum, Brassica juncea, and Litchi chinensis Honeys from Bangladesh

    Directory of Open Access Journals (Sweden)

    Nandita Sarker

    2015-01-01

    Full Text Available The present study was undertaken to determine the heavy metal levels and the physicochemical parameters (pH, electrical conductivity (EC, and ash, moisture, and total sugar content of honeys from Bangladesh. Three different floral honeys were investigated, namely, khalsi (Aegiceras corniculatum, mustard (Brassica juncea, and litchi (Litchi chinensis honeys. The heavy metals in the honeys were determined by using a High Temperature Dry Oxidation method followed by Atomic Absorption Spectroscopy. The mean pH, EC, and ash, moisture, and total sugar contents of the investigated honeys were 3.6, 0.51 mS/cm, 0.18%, 18.83%, and 68.30%, respectively. Iron was the most abundant among all the investigated heavy metals, ranging from 13.51 to 15.44 mg/kg. The mean concentrations of Mn and Zn in the investigated honeys were 0.28 mg/kg and 2.99 mg/kg, respectively. Cd was below the detection limit, and lead was found in some honey samples, but their contents were below the recommended Maximum Acceptable Level. Cr was also found in all of the samples, but its concentration was within the limit. The physicochemical analysis of the honey samples yielded levels within the limits set by the international honey legislation, indicating that the honey samples were of good quality and had acceptable values for maturity, purity, and freshness.

  4. Isolation and characterization of juncin, an antifungal protein from seeds of Japanese Takana (Brassica juncea Var. integrifolia).

    Science.gov (United States)

    Ye, Xiujuan; Ng, Tzi Bun

    2009-05-27

    An 18.9 kDa antifungal protein designated juncin was isolated from seeds of the Japanese takana (Brassica juncea var. integrifolia). The purification protocol employed comprised anion-exchange chromatography on Q-Sepharose, affinity chromatography on Affi-gel blue gel, cation exchange chromatography on SP-Sepharose, and gel filtration on Superdex 75. Juncin was adsorbed on Affi-gel blue gel and SP-Sepharose but unadsorbed on Q-Sepharose. The protein exhibited antifungal activity toward the phytopathogens Fusarium oxysporum, Helminthosporium maydis, and Mycosphaerella arachidicola with IC(50) values of 13.5, 27, and 10 μM, respectively. It was devoid of mitogenic activity toward splenocytes and nitric oxide inducing activity toward macrophages. It inhibited the proliferation of hepatoma (HepG2) and breast cancer (MCF7) cells with IC(50) values of 5.6 and 6.4 μM, respecitvely, and the activity of HIV-1 reverse transcriptase with an IC(50) of 4.5 μM. Its N-terminal sequence differed from those of antifungal proteins that have been reported to date. Compared with Brassica campestris and Brassica alboglabra antifungal peptides, juncin exhibits a different molecular mass and N-terminal amino acid sequence but similar biological activities.

  5. PENGARUH MEDIA TANAM DAN NUTRISI TERHADAP PERTUMBUHAN DAN HASIL TANAMAN PAKCHOI (Brassica juncea L. DENGAN SISTEM HIDROPONIK

    Directory of Open Access Journals (Sweden)

    Balia Perwitasari

    2012-03-01

    Full Text Available Pakchoi (Brassica juncea L.. is one kind of a vegetable crop that has high economic value and high nutrition. The technology of hydroponic is one of alternative cultivation using other media than soil substrates and nutrients. The purpose of this research is to determine the effect of media composition and nutrition on growth and yield of  pakchoi crops and find out which treatment is best for plant growth and yield of pakchoi. Researchcarried outinthe gardenexperiment ina plastichouse, Faculty ofAgriculture, University ofTrunojoyoMadura, at an altitude of±5 mabove sea level, temperature29ºC, and± 75% RH. The research was conductedin December2011 toFebruary2012.The analysis was usednonfaktorialRAL. Treatmentwithcomposemediatypes(raw rice husk, rice huskcharcoal, sand andnutrients(no nutrients, premiumnutrition, nutritiongoodplant. There are ninetreatment, threereplicationsandthreesamples ofthe plant, furthertestswere analyzed byDMRT5%. Results showedthe treatmentcompositionandnutrientmediaprovidesignificantly differentresultsatdifferent ages ofobservationson eachvariableobservation. The besttreatmentcompositioncontained in thehusk charcoalandnutrientmediagoodplant(M2N2. Evidenced by theaverage ofthe highestresultson the length ofthe plant(29.38cm, number of leaves(22.22 strands, leaf area(3226.79 cm2, wetweight(242.19 g and dry weight(13, 27g totalplantpakchoiat age4 MST.

  6. Serpentine bacteria influence metal translocation and bioconcentration of Brassica juncea and Ricinus communis grown in multi-metal polluted soils

    Science.gov (United States)

    Ma, Ying; Rajkumar, Mani; Rocha, Inês; Oliveira, Rui S.; Freitas, Helena

    2015-01-01

    The aim of this study was to assess the effects of inoculation of rhizosphere or endophytic bacteria (Psychrobacter sp. SRS8 and Pseudomonas sp. A3R3, respectively) isolated from a serpentine environment on the plant growth and the translocation and accumulation of Ni, Zn, and Fe by Brassica juncea and Ricinus communis on a multi-metal polluted serpentine soil (SS). Field collected SS was diluted to 0, 25, 50, and 75% with pristine soil in order to obtain a range of heavy metal concentrations and used in microcosm experiments. Regardless of inoculation with bacteria, the biomass of both plant species decreased with increase of the proportion of SS. Inoculation of plants with bacteria significantly increased the plant biomass and the heavy metal accumulation compared with non-inoculated control in the presence of different proportion of SS, which was attributed to the production of plant growth promoting and/or metal mobilizing metabolites by bacteria. However, SRS8 showed a maximum increase in the biomass of the test plants grown even in the treatment of 75% SS. In turn, A3R3 showed maximum effects on the accumulation of heavy metals in both plants. Regardless of inoculation of bacteria and proportion of SS, both plant species exhibited low values of bioconcentration factor (communis in SS soils and have a great potential to be used as inoculants in phytostabilization scenarios of multi-metal contaminated soils. PMID:25601876

  7. Low level of selenium increases the efficacy of 24-epibrassinolide through altered physiological and biochemical traits of Brassica juncea plants.

    Science.gov (United States)

    Naz, Fatima Salva; Yusuf, Mohammad; Khan, Tanveer A; Fariduddin, Qazi; Ahmad, Aqil

    2015-10-15

    This study was conducted to provide an insight into the effect of Se (through soil) induced changes in Brassica juncea plants in the presence and absence of 24-epibrassinolide (EBL; foliar). The Se treatments showed dual response, 10 μM of Se significantly increased growth, water relations, photosynthetic attributes along with carbonic anhydrase activity whereas its higher concentrations proved inhibitory in concentration dependent manner. The follow-up application of EBL to the Se stressed plants improved growth, water relations, photosynthesis and simultaneously enhanced the various antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase with the excess accumulation of proline. In addition to this, 10 μM Se increases the efficacy of 10(-8) M of EBL and both in combination showed maximum increase for the growth and photosynthetic traits of plants. On the other hand, the elevated level of antioxidant enzymes as well as proline could have conferred tolerance to the Se-stressed plants resulting in improved growth, water relations and photosynthesis.

  8. Effect of feeding Murraya koeingii and Brassica juncea diet on [correction] kidney functions and glucose levels in streptozotocin diabetic mice.

    Science.gov (United States)

    Grover, J K; Yadav, S P; Vats, V

    2003-03-01

    Purpose of the study was to investigate the effects of daily oral feeding 15% of powdered leaves of Murraya koeingii (MK) (commonly called as Curry patta) and 10% powder of seeds of Brassica juncea (BJ) (commonly called as Rai) for 60 days on serum glucose concentrations and kidney functions in streptozotocin (STZ; 100mg/kg) diabetic rats. Serum glucose levels, body weight, urine volume, serum creatinine, and urinary albumin (UAE) levels were monitored on day 0, 10, 25, 40, and 70 of the experiment. After 60 days of STZ administration, urine volume per day and UAE levels were significantly higher (Pdiabetic controls (DC) as compared to normal controls (NC). Although feeding of the MK/BJ showed a trend towards improvement in most of the parameters, results were not statistically different from the DC except in serum creatinine values in BJ-fed rats on day 70. Thus, these plants can be best utilized by promoting them as preferable food adjuvants for diabetic patients.

  9. Radiative transfer effects in primordial hydrogen recombination

    CERN Document Server

    Ali-Haïmoud, Yacine; Hirata, Christopher M

    2010-01-01

    The calculation of a highly accurate cosmological recombination history has been the object of particular attention recently, as it constitutes the major theoretical uncertainty when predicting the angular power spectrum of Cosmic Microwave Background anisotropies. Lyman transitions, in particular the Lyman-alpha line, have long been recognized as one of the bottlenecks of recombination, due to their very low escape probabilities. The Sobolev approximation does not describe radiative transfer in the vicinity of Lyman lines to a sufficient degree of accuracy, and several corrections have already been computed in other works. In this paper, the impact of some previously ignored radiative transfer effects is calculated. First, the effect of Thomson scattering in the vicinity of the Lyman-alpha line is evaluated, using a full redistribution kernel incorporated into a radiative transfer code. The effect of feedback of distortions generated by the optically thick deuterium Lyman-alpha line blueward of the hydrogen ...

  10. A high recombination frequency within the chicken major histocompatibility (B) complex.

    Science.gov (United States)

    Hepkema, B G; Tilanus, M G; Blankert, H J; Albers, G A; Grosfeld-Stulemeyer, M C; Hensen, E J

    1993-10-01

    Chickens of a commercial pure White Leghorn line were typed for B-F and B-G by serological, biochemical and molecular biological methods. Amongst 287 typed animals of one particular line, three animals with recombinant haplotypes were identified. Compared to earlier reports this revealed a statistically significant (P recombination frequency in this chicken line.

  11. Biochemical characterization of laccase from hairy root culture of Brassica juncea L. and role of redox mediators to enhance its potential for the decolorization of textile dyes.

    Science.gov (United States)

    Telke, Amar A; Kagalkar, Anuradha N; Jagtap, Umesh B; Desai, Neetin S; Bapat, Vishwas A; Govindwar, Sanjay P

    2011-12-01

    In vitro transgenic hairy root cultures provide a rapid system for physiological, biochemical studies and screening of plants for their phytoremediation potential. The hairy root cultures of Brassica juncea L. showed 92% decolorization of Methyl orange within 4 days. Out of the different redox mediators that were used to achieve enhanced decolorization, 2, 2'-Azinobis, 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) was found to be the most efficient. Laccase activity of 4.5 U mg(-1) of protein was observed in hairy root cultures of Brassica juncea L., after the decolorization of Methyl orange. Intracellular laccase produced by B. juncea root cultures grown in MS basal medium was purified up to 2.0 fold with 6.62 U mg(-1) specific activity using anion-exchange chromatography. Molecular weight of the purified laccase was estimated to be 148 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified enzyme efficiently oxidized ABTS which was also required for oxidation of the other tested substrates. The pH and temperature optimum for laccase activity were 4.0 and 40°C, respectively. The purified enzyme was stable up to 50°C and was stable in the pH range of 4.0-6.0. Laccase activity was strongly inhibited by sodium azide, EDTA, dithiothreitol and L: -cysteine. The purified enzyme decolorized various textile dyes in the presence of ABTS as an efficient redox mediator. These findings contribute to a better understanding of the enzymatic process involved in phytoremediation of textile dyes by using hairy roots.

  12. Effect of exogenous H2O2 on antioxidant enzymes of Brassica juncea L. seedlings in relation to 24-epibrassinolide under chilling stress.

    Science.gov (United States)

    Kumar, Manish; Sirhindi, Geetika; Bhardwaj, Renu; Kumar, Sandeep; Jain, Gagandeep

    2010-12-01

    Hydrogen peroxide is most stable molecule among reactive oxygen species, which play a vital role in growth and development of plant as signaling molecule at low concentration in response to various abiotic and biotic stresses. Exogenous application of H2O2 is known to induce chilling tolerance in plants. Brassinosteroids are plant steroid hormones known for their anti-stress properties. In this study, effect of exogenous H2O2 on antioxidant defense system of Brassica juncea L. seedlings was investigated in 24-epibrassinolide (24-EBL) treated and untreated seedlings under chilling stress. The surface sterilized seeds of B. juncea L. were germinated in petriplates containing different concentrations of H2O2 alone and in combination with 10(-8) M 24-EBL. Chilling treatment (4 degrees C) was given to 10-days old seedlings grown in different treatments for 6 h daily up to 3 days. 24 h recovery period was given to chilling treated seedlings by placing at 25 degrees C + 2 degrees C and harvested for antioxidant enzymes on 14th day after sowing (DAS). Treatment of 24-EBL in combination with H2O2 (15 and 20 mM) helped in reducing the toxicity of seed and seedlings due to H2O2 exposure on their germination rate, shoot and root length respectively. 24-EBL treatment at seed and seedling stage helped in alleviating the toxic effect of H2O2 through antioxidant defense system by increasing the activities of various enzymes involved in antioxidant defense system such as catalase (CAT, E.C. 1.11.1.6), ascorbate peroxidase (APOX, E.C. 1.11.1.11), and superoxide dismutase (SOD, E.C. 1.15.1.1). In conclusion, exogenous pretreatment of H2O2 to seeds of B. juncea L. adapted the seedlings to tolerate chilling stress, which was further ameliorated in combination of H2O2 with 24-EBL.

  13. Global insights into high temperature and drought stress regulated genes by RNA-Seq in economically important oilseed crop Brassica juncea.

    Science.gov (United States)

    Bhardwaj, Ankur R; Joshi, Gopal; Kukreja, Bharti; Malik, Vidhi; Arora, Priyanka; Pandey, Ritu; Shukla, Rohit N; Bankar, Kiran G; Katiyar-Agarwal, Surekha; Goel, Shailendra; Jagannath, Arun; Kumar, Amar; Agarwal, Manu

    2015-01-21

    Brassica juncea var. Varuna is an economically important oilseed crop of family Brassicaceae which is vulnerable to abiotic stresses at specific stages in its life cycle. Till date no attempts have been made to elucidate genome-wide changes in its transcriptome against high temperature or drought stress. To gain global insights into genes, transcription factors and kinases regulated by these stresses and to explore information on coding transcripts that are associated with traits of agronomic importance, we utilized a combinatorial approach of next generation sequencing and de-novo assembly to discover B. juncea transcriptome associated with high temperature and drought stresses. We constructed and sequenced three transcriptome libraries namely Brassica control (BC), Brassica high temperature stress (BHS) and Brassica drought stress (BDS). More than 180 million purity filtered reads were generated which were processed through quality parameters and high quality reads were assembled de-novo using SOAPdenovo assembler. A total of 77750 unique transcripts were identified out of which 69,245 (89%) were annotated with high confidence. We established a subset of 19110 transcripts, which were differentially regulated by either high temperature and/or drought stress. Furthermore, 886 and 2834 transcripts that code for transcription factors and kinases, respectively, were also identified. Many of these were responsive to high temperature, drought or both stresses. Maximum number of up-regulated transcription factors in high temperature and drought stress belonged to heat shock factors (HSFs) and dehydration responsive element-binding (DREB) families, respectively. We also identified 239 metabolic pathways, which were perturbed during high temperature and drought treatments. Analysis of gene ontologies associated with differentially regulated genes forecasted their involvement in diverse biological processes. Our study provides first comprehensive discovery of B. juncea

  14. 26. 13-MHz absorption line toward Cassiopeia A

    Energy Technology Data Exchange (ETDEWEB)

    Konovalenko, A.A.; Sodin, L.G.

    1981-07-01

    New observations confirm that the 26.13-MHz absorption line previously detected in the direction toward the radio source Cas A represents a carbon recombination line, not a hyperfine-structure nitrogen line. Some relevant implications are discussed.

  15. A Nestin-cre transgenic mouse is insufficient for recombination in early embryonic neural progenitors

    Directory of Open Access Journals (Sweden)

    Huixuan Liang

    2012-09-01

    Nestin-cre transgenic mice have been widely used to direct recombination to neural stem cells (NSCs and intermediate neural progenitor cells (NPCs. Here we report that a readily utilized, and the only commercially available, Nestin-cre line is insufficient for directing recombination in early embryonic NSCs and NPCs. Analysis of recombination efficiency in multiple cre-dependent reporters and a genetic mosaic line revealed consistent temporal and spatial patterns of recombination in NSCs and NPCs. For comparison we utilized a knock-in Emx1cre line and found robust recombination in NSCs and NPCs in ventricular and subventricular zones of the cerebral cortices as early as embryonic day 12.5. In addition we found that the rate of Nestin-cre driven recombination only reaches sufficiently high levels in NSCs and NPCs during late embryonic and early postnatal periods. These findings are important when commercially available cre lines are considered for directing recombination to embryonic NSCs and NPCs.

  16. Monitoring homologous recombination in rice (Oryza sativa L.)

    Energy Technology Data Exchange (ETDEWEB)

    Yang Zhuanying; Tang Li [Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou 510631 (China); Li Meiru [South China Botanic Garden, Chinese Academy of Sciences, Guangzhou 510650 (China); Chen Lei; Xu Jie [Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou 510631 (China); Wu Goujiang [South China Botanic Garden, Chinese Academy of Sciences, Guangzhou 510650 (China); Li Hongqing, E-mail: hqli@scnu.edu.cn [Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou 510631 (China)

    2010-09-10

    Here we describe a system to assay homologous recombination during the complete life cycle of rice (Oryza sativa L.). Rice plants were transformed with two copies of non-functional GUS reporter overlap fragments as recombination substrate. Recombination was observed in all plant organs examined, from the seed stage until the flowering stage of somatic plant development. Embryogenic cells exhibited the highest recombination ability with an average of 3 x 10{sup -5} recombination events per genome, which is about 10-fold of that observed in root cells, and two orders of that observed in leaf cells. Histological analysis revealed that recombination events occurred in diverse cell types, but preferentially in cells with small size. Examples of this included embryogenic cells in callus, phloem cells in the leaf vein, and cells located in the root apical meristem. Steady state RNA analysis revealed that the expression levels of rice Rad51 homologs are positively correlated with increased recombination rates in embryogenic calli, roots and anthers. Finally, radiation treatment of plantlets from distinct recombination lines increased the recombination frequency to different extents. These results showed that homologous recombination frequency can be effectively measured in rice using a transgene reporter assay. This system will facilitate the study of DNA damage signaling and homologous recombination in rice, a model monocot.

  17. Organic matrix based slow release fertilizer enhances plant growth, nitrate assimilation and seed yield of Indian mustard (Brassica juncea L.).

    Science.gov (United States)

    Sharma, Vinod K; Singh, Rana P

    2011-09-01

    Field experiments were conducted to study the effect of organic matrix based slow release fertilizers (SRFs) on plant growth, nitrate assimilation and seed yield of Brassica juncea L. cv, pusa bold. The agro-waste materials like cow dung, clay soil, neem leaves and rice bran were mixed together in 2:2:1:1 ratio and used as organic matrix for the immobilization of chemical fertilizer nutrients with commercial grade saresh (Acacia gum, 15% solution) as binder. Different fertilizer treatments were organic matrix based slow release fertilizers, SRF-I (542.0 kg ha(-1)); SRF-II (736.5 kg ha(-1)) and chemical fertilizer combinations, boron (3 kg ha(-1))+sulphur (15 kg ha(-1))+nitrogen (80 kg ha(-1)) and boron (3 kg ha(-1)) + sulphur (15 kg ha(-1))+nitrogen (80 kg ha(-1))+phosphorus (15 kg ha(-1))+potassium (100 kg ha(-1)). Organic matrix based SRF-II released ammonium up to 50-d in wetsoil under laboratory conditions which showed maximum retention of the nutrients. Avery significant increase in plant growth, nitrate assimilation and seed yield was recorded in organic matrix based SRF-II applied plants. The maximum percent increase in biomass production was observed with organic matrix based SRF-II (increase of 65.8% in root fresh weight, 38.0% in root dry weight, 45.9% in leaf fresh weight plant(-1) and 27.5 % in leaf dry weight plant(-1) in 60-d old plants). It also increased the acquisition and assimilation of nitrate from the plant's rhizosphere which was evident by 45.6% increase in nitrate, 27.5% in nitrite and 11.7% in nitrate reductase activity (NRA) in leaves of 45-d old plants over control. The organic matrix based SRF-II significantly increased the seed yield by 28% in Indian mustard. Cost analysis revealed thatthis formulation is cost effective as it is based on agro waste materials.

  18. Alleviation of cadmium toxicity in Brassica juncea L. (Czern. & Coss. by calcium application involves various physiological and biochemical strategies.

    Directory of Open Access Journals (Sweden)

    Parvaiz Ahmad

    Full Text Available Calcium (Ca plays important role in plant development and response to various environmental stresses. However, its involvement in mitigation of heavy metal stress in plants remains elusive. In this study, we examined the effect of Ca (50 mM in controlling cadmium (Cd uptake in mustard (Brassica juncea L. plants exposed to toxic levels of Cd (200 mg L(-1 and 300 mg L(-1. The Cd treatment showed substantial decrease in plant height, root length, dry weight, pigments and protein content. Application of Ca improved the growth and biomass yield of the Cd-stressed mustard seedlings. More importantly, the oil content of mustard seeds of Cd-stressed plants was also enhanced with Ca treatment. Proline was significantly increased in mustard plants under Cd stress, and exogenously sprayed Ca was found to have a positive impact on proline content in Cd-stressed plants. Different concentrations of Cd increased lipid peroxidation but the application of Ca minimized it to appreciable level in Cd-treated plants. Excessive Cd treatment enhanced the activities of antioxidant enzymes superoxide dismutase, ascorbate peroxidase and glutathione reductase, which were further enhanced by the addition of Ca. Additionally, Cd stress caused reduced uptake of essential elements and increased Cd accumulation in roots and shoots. However, application of Ca enhanced the concentration of essential elements and decreased Cd accumulation in Cd-stressed plants. Our results indicated that application of Ca enables mustard plant to withstand the deleterious effect of Cd, resulting in improved growth and seed quality of mustard plants.

  19. Application of bokashi and sunn hemp (Crotalaria juncea L. to improve inorganic fertilizer efficiency on maize (Zea mays L.

    Directory of Open Access Journals (Sweden)

    A.I. Yuliana

    2015-10-01

    Full Text Available A field experiment was conducted lo learn about the effect of Bokashi and Sunn hemp (Crotalaria juncea L. on maize production and inorganic fertilizer use efficiency on maize. The experiment was conducted in Jatikerto, Malang; at the altitude of 303 m above sea level, in Alfisol soil type, the average daily temperature ranges 21-33oC, from June to October 2013. The experiment was conducted as factorial, designed in a randomized block design (RBD. The first factor was dose of inorganic fertilizer (100% ; 75% and 50% of recommendation dose. The second factor was the organic fertilizer (Without organic fertilizer20 t Bokashi/ ha, 20 t Sunn hemp/ha, 10 t Bokashi/ha + 10 t Sunn hemp/ha. The results showed that application of 20 t Bokashi/ha, 20 t Sunn hemp/ha, and combination of 10 t Bokashi/ha + 10 t Sunn hemp/ha, along with the application of inorganic fertilizer by dose of 100% increased the yields of maize for about 41.8%; 47.6% and 54.7% (10.73 t/ha; 11.17 t/ha, and 11.71 t/ha, respectively. The yield and nutrient use efficiency in the treatment dose of 100% inorganic fertilizer did not have any significant difference from the application of 20 t Bokashi /ha, 20 t Sunn hemp/ha, and 10 t Bokashi/ha + 10 t Sunn hemp/ha along with doses of inorganic fertilization 75% and 50%. Therefore, the organic fertilizer of 20 t Bokashi/ ha, 20 t Sunn hemp/ha, and combination of 10 t Bokashi/ha + 10 t Sunn hemp/ha could reduce the need of inorganic fertilizer for about 50%.

  20. Influence of indian mustard (Brassica juncea) on rhizosphere soil solution chemistry in long-term contaminated soils: a rhizobox study.

    Science.gov (United States)

    Kim, Kwon-Rae; Owens, Gary; Kwon, Soon-lk

    2010-01-01

    This study investigated the influence of Indian mustard (Brassica juncea) root exudation on soil solution properties (pH, dissolved organic carbon (DOC), metal solubility) in the rhizosphere using a rhizobox. Measurement was conducted following the cultivation of Indian mustard in the rhizobox filled four different types of heavy metal contaminated soils (two alkaline soils and two acidic soils). The growth of Indian mustard resulted in a significant increase (by 0.6 pH units) in rhizosphere soil solution pH of acidic soils and only a slight increase (alkaline soils. Furthermore, the DOC concentration increased by 17-156 mg/L in the rhizosphere regardless of soil type and the extent of contamination, demonstrating the exudation of DOC from root. Ion chromatographic determination showed a marked increase in the total dissolved organic acids (OAs) in rhizosphere. While root exudates were observed in all soils, the amount of DOC and OAs in soil solution varied considerably amongst different soils, resulting in significant changes to soil solution metals in the rhizosphere. For example, the soil solution Cd, Cu, Pb, and Zn concentrations increased in the rhizosphere of alkaline soils compared to bulk soil following plant cultivation. In contrast, the soluble concentrations of Cd, Pb, and Zn in acidic soils decreased in rhizosphere soil when compared to bulk soils. Besides the influence of pH and DOC on metal solubility, the increase of heavy metal concentration having high stability constant such as Cu and Pb resulted in a release of Cd and Zn from solid phase to liquid phase.

  1. Early osmotic, antioxidant, ionic, and redox responses to salinity in leaves and roots of Indian mustard (Brassica juncea L.).

    Science.gov (United States)

    Ranjit, Singh Laxmi; Manish, Pandey; Penna, Suprasanna

    2016-01-01

    Salt-stress-induced alterations in osmotic, ionic, and redox responses were studied in the early period of treatment (30 min to 5 days) in seedlings of Brassica juncea L. Roots and shoots under mild (50 mM) and severe (250 mM) NaCl stress were analyzed for growth, oxidative stress, osmolyte accumulation, antioxidant defense, and redox state. Growth reduction was less pronounced in the early time period of salt stress while oxidative damage increased linearly and in a sustained manner under severe stress up to 6 h. An early and transient reactive oxygen species (ROS) burst, as evidenced by superoxide and hydrogen peroxide level was observed, followed by activation of enzymatic antioxidant system (GPX, SOD, CAT, and GR) in both root and shoot. The enzymatic activity was not affected much under mild stress particularly at early phase; however, severe stress induced a significant increase in the activity of antioxidant enzymes. Root ascorbate was progressively accumulated, and its redox state maintained in the early time phase of treatment under mild stress while increase in root and shoot glutathione content was recorded under mild stress at 5 days when the active ascorbate pool decreased. While early period of salt stress showed significant Na(+) accumulation over control, plants subjected to mild stress measured less Na(+) accumulation up to 5 days compared to severely stressed plants. The results showed an early induction of differential responses to salt stress in roots and shoots of Brassica which include growth limitations, reduced relative water content, increased osmolytes, redox state, and antioxidant system, and a significant Na(+) increase. The results also indicate that roots and shoots may have distinct mechanisms of responses to salt stress.

  2. Novel Recombinant Sapovirus

    Science.gov (United States)

    Katayama, Kazuhiko; Miyoshi, Tatsuya; Uchino, Kiyoko; Oka, Tomoichiro; Tanaka, Tomoyuki; Takeda, Naokazu

    2004-01-01

    We determined the complete genome sequences of two sapovirus strains isolated in Thailand and Japan. One of these strains represented a novel, naturally occurring recombinant sapovirus. Evidence suggested the recombination site was at the polymerase-capsid junction within open reading frame one. PMID:15504283

  3. 利用重组自交系群体构建番茄AFLP遗传连锁图谱%Construction AFLP Genetic Linkage Map of Tomato Using Recombinant Inbred Lines (RILs) Population

    Institute of Scientific and Technical Information of China (English)

    陈丽静; 王利; 王玉坤; 陶承光; 李君明; 王晓武; 李天来

    2012-01-01

    以普通栽培番茄(Solanum lycopersicum)99165.30为母本,野生多毛番茄(Solanumhabrochaites)LAl777为父本进行杂交,通过单粒传得到了含有80个F5:6家系的重组自交系分离群体,利用荧光AFLP分子标记技术构建番茄分子遗传连锁图谱。AFLP标记采用MseI和EcoRI两种内切酶及荧光标记(IRD.700或IRD.800)的E+3和非荧光标记的M+3引物组合进行选择性扩增,扩增结果经95℃预变性后在6%变性聚丙烯酰胺凝胶上电泳2.5h,运用LICOR公司的NENGlobal Edition IR2 DNA Analyzer(Model 5200 LI-COR Biosciences,Lincoln,NE)荧光扫描检测DNA多态性。对RILs群体中产生分离的274个AFLP标记运用Join Map3.0软件分析,得到一张番茄分子遗传连锁图谱,图谱总长度为662cM,共包括18个主要连锁群,125个多态性分子标记。每条连锁群上的标记数在3~22个之间,连锁群的长度在14.0~58.0cM的范围内,平均图距在2-27~13-3cM。总平均距离5.3cM,本研究中构建的番茄永久遗传图谱,为番茄分子辅助育种及重要农艺性状的定位奠定了基础。%A genetic linkage map of tomato was constructed using a RILs (recombinant inbred lines) population of 80 individuals which was developed by crossing Solanum lycopersicum 99165-30 and Solanum habrochaites LA1777 through single-seed descent (SSD) . AFLPs were generated by the use of restriction enzymes EcoR I in combination with either Mse I. Pre-amplification was carded out using primers corresponding to EcoR I and Mse I adaptors with no selective base. Selective amplifications were performed using IRD700 or IRD800 labeled EcoR I primers and non-labeled Mse I primers. The resulting products were denatured in formamide at 95℃ and separated by electrophoresis 2.5 h on 6% polyacrylamide gel using IR2 DNA Analyzer (Model 5200 LI-COR Biosciences, Lincoln, NE) . The segregation of each marker and linkage analysis was done

  4. Uptake of heavy metals and As by Brassica juncea grown in a contaminated soil in Aznalcóllar (Spain): the effect of soil amendments.

    Science.gov (United States)

    Clemente, Rafael; Walker, David J; Bernal, M Pilar

    2005-11-01

    Two crops of Brassica juncea (L.) Czern. were grown in a field experiment, at the site affected by the toxic spillage of acidic, metal-rich waste in Aznalcóllar (Seville, Spain), to study its metal accumulation and the feasibility of its use for metal phytoextraction. The effects of organic soil amendments (cow manure and mature compost) and lime on biomass production and plant survival were also assessed; plots without organic amendment and without lime were used as controls. Plots, with or without organic amendment, having pH Soil acidification conditioned plant growth and metal accumulation. The addition of lime and the organic amendments achieved higher plant biomass production, although effects concerning metal bioavailability and accumulation were masked somewhat by pH variability with time and between and within plots. Tissue metal concentrations of B. juncea were elevated for Zn, Cu and Pb, especially in leaves of plants from plots with low pH values (maxima of 2029, 71 and 55 microg g(-1), respectively). The total uptake of heavy metals in the plants was relatively low, emphasising the problems faced when attempting to employ phytoextraction for clean-up of pluri-contaminated sites.

  5. Kajian Hubungan Populasi Tanaman dengan Neraca Unsur Hara Nitrogen dan Fosfor pada Sistem Vertikultur Sawi Hijau (Brassica juncea L dan Kangkung (Ipomea reptana

    Directory of Open Access Journals (Sweden)

    I MADE PURNA WIDANA

    2016-05-01

    Full Text Available Analysis of Relationship Between Crop Density and The Balancing of Both Soil Nitrogenand Phosporus in Verticulture System of Mustard Green (Brassica juncea L and Kale(Ipomea reptana. The relationship between crops density and the balancing of both soilnitrogen and phosphorus in verticulture system remained to be important issue. A glasshouseexperiment had carried out from October 2014 to March 2015 in order to determine (1 theoptimum population density of both mustard greens (Brassica juncea L and kale (Ipomeareptana, (2 soil total-N or available-P balance, and (3 the relatiohships between plantspopulations and soil nutrients balance. A split plot experiment under complete block designwas applied to examine the effect of the main plot (crops type i.e. mustard green and kale andsub plot (crops population i.e 10, 15, 20,25, and 30 crops per planting container 0,12 m2 insize. The results showed that no optimal crops population density had achieved. Themaximum crops population was 30 for both mustard greens and kale. The soil total-N balancewas negative while these was positive for soil available-P balance of P and N negative. Alogarithmic relationships was calculated between soil total-N balance with mustard green,while linier patterns were significant for soil-N balance with kale and available-P balancewith both mustard greens and kale.

  6. Pre-sowing Seed Treatment with 24-Epibrassinolide Ameliorates Pesticide Stress in Brassica juncea L. through the Modulation of Stress Markers

    Directory of Open Access Journals (Sweden)

    Parvaiz Ahmad

    2016-11-01

    Full Text Available The present experiment was designed to assess the effects of seed soaking with 24-epibrassinolide (EBR on the physiology of Brassica juncea L. seedlings grown under imidacloprid (IMI toxicity. Application of EBR increased the length of seedlings, dry weight, and pigment contents, polyphenols, total phenols and organic acids under IMI toxicity. The expression of genes coding key enzymes of pigment, phenols, polyphenols and organic acid biosynthetic pathways was also studied including CHLASE (chlorophyllase, PSY (phytoene synthase, CHS (chalcone synthase and PAL (phenylalanine ammonialyase, CS (citrate synthase, SUCLG1 (succinyl Co-A ligase,, SDH (succinate dehydrogenase, FH (fumarate hydratase, MS (malate synthase. Multiple linear regression analysis revealed that IMI application regressed negatively on seedling length, dry weight and total chlorophyll content. However, EBR seed treatment regressed positively on all of the parameters studied. Moreover, interaction between IMI and EBR showed positive regression for growth parameters, content of pigments, total polyphenol, total phenol and malate, and expression of PSY and PAL. Negative interactions were noticed for the contents of fumarate, succinate and citrate, and expression of CHS and all genes studied related to organic acid metabolism. In conclusion, EBR enhanced the growth and contents of all studied metabolites by regulating the gene expression of B. juncea seedlings under IMI stress.

  7. Effect of seed pre-soaking with 24-epibrassinolide on growth and photosynthetic parameters of Brassica juncea L. in imidacloprid soil.

    Science.gov (United States)

    Sharma, Anket; Kumar, Vinod; Singh, Ravinder; Thukral, Ashwani Kumar; Bhardwaj, Renu

    2016-11-01

    Pesticides are widely used to protect crop plants from various insect pests. However, application of pesticides causes phytotoxicity to plants which results in their impaired growth and development. Brassinosteroids are well known to protect plants under abiotic stress conditions. The purpose of the present study was to access the ameliorative role of 24-epibrassinolide (EBR) in Brassica juncea L. under imidacloprid (IMI) toxicity. B. juncea plants were raised from seeds soaked in 0.1, 1 and 100nM of EBR, and grown in soils amended with 250, 300 and 350mgkg(-1) IMI pesticide, and observed for growth, pigments and photosynthetic parameters after 30, 60 and 90 days of seed sowing. The plants grown in soil treated with IMI exhibited a significant reduction in shoot length, number of leaves, chlorophyll contents and photosynthetic parameters like photosynthetic rate, stomatal conductance, inter-cellular CO2 and transpiration rate, when compared with their respective controls. However, pigments which act as antioxidants such as carotenoids, anthocyanins and xanthophylls were increased with IMI stress. Pre-sowing seed treatment with EBR decreased the toxic effects of IMI and increased the growth, pigment biosynthesis and photosynthetic parameters of the plants grown in IMI amended soil. Maximum increase in all the growth and photosynthetic parameters was noticed in plants raised from seeds treated with 100nM EBR and grown in IMI amended soil.

  8. Insecticidal, food utilisation and biochemical effect of essential oils extracted from seeds of Brassica juncea (Czern. against Spodoptera litura (Lepidoptera: Noctuidae (Fabricius

    Directory of Open Access Journals (Sweden)

    Shallina Gupta

    2017-09-01

    Full Text Available The glucosinolate rich hot and cold hexane extracts of Brassica juncea had a negative effect on the development of Spodoptera litura larvae when they were fed on different concentrations of the extracts. Both larval growth index and pupal growth index declined with treatment. However the hot extract with 3-butenyl isothiocyanate as the predominant compound had a more deleterious effect as at 3125ppm the larvae were unable to complete their development. The nutritional indices too were more adversely affected with hot extract as compared to cold extract. The RGR, RCR, ECI and ECD declined significantly with increase in concentration of the hot hexane extract. The activity of antioxidant enzymes, SOD and catalase decreased while that of phosphatases, GSTs, phenol oxidases increased during the initial treatment duration but decreased on prolonged treatment of the larvae with LC50 concentration of hot extract. A similar trend was observed for glutathione and lipid peroxides but a decrease in ascorbate content was observed as compared to control. The findings reveal a toxic effect of 3-butenylisothiocyanate rich hot hexane extract of B. juncea on S. litura larvae.

  9. Recombinant methods and materials

    Energy Technology Data Exchange (ETDEWEB)

    Roizman, B.; Post, L.E.

    1988-09-06

    This patent describes a method for stably effecting the insertion or deletion of a selected DNA sequence at a specific site in a viral genome. The method consists of: (1) isolating from the genome a linear DNA fragment comprising both (a) the specific site determined for insertion or deletion of selected DNA sequence and (b) flanking DNA sequences normally preceding and following the site; (2) preparing first and second altered genome fragments from the fragment isolated in step (1). (a) the first altered fragment comprising the fragment comprising a thymidine kinase gene in a position intermediate the ends of the fragment, and (b) the second altered fragment comprising the fragment having the selected DNA sequence inserted therein or deleted therefrom; (3) contacting the genome with the first altered fragment under conditions permitting recombination at sites of DNA sequence homology, selecting for a recombinant genome comprising the thymidine kinase gene, and isolating the recombinant genome; and (4) contacting the recombinant genome isolated in step (3) with the second altered fragment under conditions permitting recombination at sites of DNA sequence homology, selecting for a recombinant genome lacking the thymidine kinase gene, and isolating the recombinant genome product.

  10. Interacción genotipo-ambiente de 135 líneas endogámicas recombinantes de sorgo para producción de biocombustibles G*E interaction of 135 recombinant inbred lines of sorghum for biofuel production

    Directory of Open Access Journals (Sweden)

    Armando Rodríguez García

    2012-02-01

    Full Text Available El sorgo (Sorghum bicolor (L Moench, es uno de los alimentos básicos para la población más pobre del mundo, que es también la que padece una situación de mayor inseguridad alimentaria. Este cultivo se adapta a un entorno agroecológico cálido y seco, donde es difícil cultivar otros cereales alimenticios. La formación de genotipos sobresalientes requiere evaluar los materiales genéticos en diferentes ambientes y determinar su interacción genotipo-ambiente. El objetivo del trabajo fue detectar y analizar la interacción genotipo-ambiente de un grupo de líneas endogámicas recombinantes y seleccionar las de mayor potencial, para ser utilizadas en la producción de biocombustibles. Se evaluaron 135 líneas endogámicas recombinantes de sorgo en diferentes localidades durante los años 2006, 2007 y 2008. Los datos de siete variables se analizaron como corresponde a un experimento en diseño de bloques al azar, en cada uno de varios ambientes. La interacción genotipo-ambiente, se analizó mediante el modelo AMMI. Fue altamente significativa, calificando las líneas endogámicas recombinantes por su magnitud y seleccionando las de menor magnitud, se determinó el mejor ambiente para el desarrollo adecuado de las líneas endogámicas recombinantes y se seleccionaron 13 líneas con mayor contenido de azúcar medido en °Bx.Sorghum (Sorghum bicolor (L Moench is one of the main basic foods forthe poorest people in the world, suffering from a situation of increased food insecurity. This crop is adapted to a warmer and drier agro-ecological environment, where it's difficult to grow other cereals. The formation of outstanding genotypes requires to evaluating the genetic materials in different environments and determines their genotype-environment interaction. The objective was to detect and analyze the genotype-environmentofagroup of recombinant inbred lines and select the highest potential to be used in the production of biofuels. 135 recombinant

  11. Study on A New Hybrid Breeding between Raphanus sativus and Brassica juncea by Distant Hybridization%萝卜与芥菜远缘杂交新品种的选育研究

    Institute of Scientific and Technical Information of China (English)

    张小康; 熊秋芳; 李世升; 张雪清; 张贵生

    2012-01-01

    In this study, the male sterile line of radish (Raphanus sativus L. ) variety named Wuqing was used as female parent, and three Brassica juncea varieties named "Xuelihong" , "Turnips" , and "Mustard" were used as male parents, respectively. The hybrid varieties were obtained through distant hybridization made by bees in greenhouse. The hybrids were planted in field and their characters were determined for identifying the true hybrid. One hybrid variety with obvious variation in leaf type character was identified here. The inbreed lines by selling segregated seven distinct types of individuals. "Luojie -4" was a stable variety among them and its characteristics including growth form, trend of root' s plunge, and root distribution were observed. Moreover, the varieties' similarity at early growth stage was analyzed. The nutrition components of "Luojie -4" and control varieties were also analyzed. Hybrid combinations were obtained successfully from the hybridization between "Wu A" and "Luojie -4". The new combinations, which had genetic stability of the leaf traits and fleshy root heterosis, were very suitable for being pickled.%以萝卜不育系为母本,雪里蕻、大头菜和多裂叶芥为父本,在温室通过蜜蜂进行远缘杂交,并获得杂交种.经过田间种植淘汰掉假杂种,获得一株叶形明显变异的品种,对其自交,其分离成7个类型品系.选取较稳定的“萝芥4号”品系观察了其生长形态、下扎趋势以及根系分布特性,分析了早期生长过程中品种间的相似性,同时针对“萝芥4号”及对照品种进行营养成分分析.采用“武A”雄性不育系与“萝芥4号”配制杂交组合,获得叶片性状遗传稳定、肉质根杂交优势明显的组合,该组合十分适合腌制加工.

  12. Advances in recombinant antibody manufacturing.

    Science.gov (United States)

    Kunert, Renate; Reinhart, David

    2016-04-01

    Since the first use of Chinese hamster ovary (CHO) cells for recombinant protein expression, production processes have steadily improved through numerous advances. In this review, we have highlighted several key milestones that have contributed to the success of CHO cells from the beginning of their use for monoclonal antibody (mAb) expression until today. The main factors influencing the yield of a production process are the time to accumulate a desired amount of biomass, the process duration, and the specific productivity. By comparing maximum cell densities and specific growth rates of various expression systems, we have emphasized the limiting parameters of different cellular systems and comprehensively described scientific approaches and techniques to improve host cell lines. Besides the quantitative evaluation of current systems, the quality-determining properties of a host cell line, namely post-translational modifications, were analyzed and compared to naturally occurring polyclonal immunoglobulin fractions from human plasma. In summary, numerous different expression systems for mAbs are available and also under scientific investigation. However, CHO cells are the most frequently investigated cell lines and remain the workhorse for mAb production until today.

  13. A recombinant CHSE-214 cell line expressing an Mx1 promoter-reporter system responds to both interferon type I and type II from salmonids and represents a versatile tool to study the IFN-system in teleost fish.

    Science.gov (United States)

    Jørgensen, Jorunn B; Johansen, Audny; Hegseth, Marit N; Zou, Jun; Robertsen, Børre; Collet, Bertrand; Secombes, Christopher J

    2007-12-01

    A transgenic cell line for the detection of salmon interferons (IFNs) has been established. It is based on a CHSE-214 cell line containing a reporter construct expressing firefly luciferase under the control of the rainbow trout promoter for the IFN-induced Mx1 gene. This cell line, named CHSE-Mx10, showed IFN-induced luciferase expression after more than 80 passages, confirming the stability of this cell line. Interestingly, the Mx promoter was shown to respond to both salmon IFN-alpha/beta and trout IFN-gamma in a dose-dependent manner, while there was no response to TNF-alpha and IL-1beta. IFN-alpha/beta activity could be measured at a range of 9-150 U/ml, and IFN-gamma showed activity between 10 and 100 ng/ml. The reproducibility of both responses was good. The CHSE-Mx10 reporter system constitutes a versatile tool to study the induction and regulation of IFN signaling in teleost fish. A preliminary study presented herein suggests that both infectious pancreas necrosis virus (IPNV) and salmon pancreas disease virus (SPDV) may block activation of the Mx promoter in CHSE-Mx10 stimulated with IFN-alpha/beta.

  14. Novel intragenotype recombination in sapovirus.

    Science.gov (United States)

    Phan, Tung Gia; Yan, Hainian; Khamrin, Pattara; Quang, Trinh Duy; Dey, Shuvra Kanti; Yagyu, Fumihiro; Okitsu, Shoko; Müller, Werner E G; Ushijima, Hiroshi

    2006-01-01

    Based on the genetic analysis, a novel, naturally occurring recombination between two distinct sapovirus subtypes (subtype a and subtype b) within genogroup I genotype 1 was identified. Breakpoint analysis of recombinant sapovirus showed that the recombination site was at the polymerase-capsid junction. This is the first report of the existence of acute gastroenteritis caused by intragenotype recombinant sapovirus. The results also provided evidence that the natural recombination occurs not only in sapovirus genogroup II but also in sapovirus genogroup I.

  15. Regulation of Meiotic Recombination

    Energy Technology Data Exchange (ETDEWEB)

    Gregory p. Copenhaver

    2011-11-09

    Meiotic recombination results in the heritable rearrangement of DNA, primarily through reciprocal exchange between homologous chromosome or gene conversion. In plants these events are critical for ensuring proper chromosome segregation, facilitating DNA repair and providing a basis for genetic diversity. Understanding this fundamental biological mechanism will directly facilitate trait mapping, conventional plant breeding, and development of genetic engineering techniques that will help support the responsible production and conversion of renewable resources for fuels, chemicals, and the conservation of energy (1-3). Substantial progress has been made in understanding the basal recombination machinery, much of which is conserved in organisms as diverse as yeast, plants and mammals (4, 5). Significantly less is known about the factors that regulate how often and where that basal machinery acts on higher eukaryotic chromosomes. One important mechanism for regulating the frequency and distribution of meiotic recombination is crossover interference - or the ability of one recombination event to influence nearby events. The MUS81 gene is thought to play an important role in regulating the influence of interference on crossing over. The immediate goals of this project are to use reverse genetics to identify mutants in two putative MUS81 homologs in the model plant Arabidopsis thaliana, characterize those mutants and initiate a novel forward genetic screen for additional regulators of meiotic recombination. The long-term goal of the project is to understand how meiotic recombination is regulated in higher eukaryotes with an emphasis on the molecular basis of crossover interference. The ability to monitor recombination in all four meiotic products (tetrad analysis) has been a powerful tool in the arsenal of yeast geneticists. Previously, the qrt mutant of Arabidopsis, which causes the four pollen products of male meiosis to remain attached, was developed as a facile system

  16. Simultaneous quantification of sinigrin, sinalbin, and anionic glucosinolate hydrolysis products in Brassica juncea and Sinapis alba seed extracts using ion chromatography.

    Science.gov (United States)

    Popova, Inna E; Morra, Matthew J

    2014-11-01

    Although mustards such as Sinapis alba and Brassica juncea contain glucosinolates (sinalbin and sinigrin, respectively) that hydrolyze to form biopesticidal products, routine quality control methods to measure active ingredients in seed and seed meals are lacking. We present a simple and fast ion chromatography method for the simultaneous quantification of sinigrin, sinalbin, and anionic hydrolysis products in mustard seed to assess biological potency. Optimum conditions include isocratic elution with 100 mM NaOH at a flow rate of 0.9 mL/min on a 4 × 210 mm hydroxide-selective anion-exchange column. All anion analytes including sinigrin, sinalbin, SO4(2-), and SCN(-) yielded recoveries ranging from 83 to 102% and limits of detection ≤0.04 mM, with samples displaying little interference from plant matrix components. Sample preparation is minimized and analysis times are shortened to <90 min as compared with previous methods that took days and multiple instruments.

  17. Study of phytochelatins and other related thiols as complexing biomolecules of As and Cd in wild type and genetically modified Brassica juncea plants.

    Science.gov (United States)

    Navaza, Ana Pereira; Montes-Bayón, Maria; LeDuc, Danika L; Terry, Norman; Sanz-Medel, Alfredo

    2006-03-01

    The accumulation of As and Cd in Brassica juncea plants and the formation of complexes of these elements with bioligands such as glutathione and/or phytochelatins (PCs) is studied. The genetic manipulation of these plants to induce higher As and Cd accumulation has been achieved by overexpressing the genes encoding for gamma-glutamyl cysteine synthetase (gamma-ECS) and glutathione synthetase (GS). These two enzymes are responsible for glutathione (GSH) formation in plants, which is the first step in the production of PCs. The biomass produced in both the wild type and the genetically modified plants, has been evaluated. Additionally, the total Cd and As concentration accumulated in the plant tissues was measured by inductively coupled plasma mass spectrometry (ICP-MS) after extraction. Speciation studies on the extracts were conducted using size exclusion liquid chromatography (SEC) coupled online with ICP-MS to monitor As, Cd and S. For further purification of the As fractions, reversed phase high performance liquid chromatography (RP-HPLC) was used. Structural elucidation of the PCs and other thiols, as well as their complexes with As and Cd, was performed by electrospray-quadrupole-time-of-flight (ESI-Q-TOF). In both the Cd and As exposed plants it was possible to observe the presence of oxidized PC2 ([M + H]+, m/z 538), GS-PC2(-Glu) ([M + H]+, m/z 716) as well as reduced GSH ([M + H]+, m/z 308) and oxidized glutathione (GSSG) ([M + H]+, m/z 613). However, only the GS plants exhibited the presence of As(GS)3 complex ([M + H]+, m/z 994) that was further confirmed by MS/MS. This species is reported for the first time in B. juncea plant tissues.

  18. Intergenogroup Recombination in Sapoviruses

    Science.gov (United States)

    Hansman, Grant S.; Takeda, Naokazu; Oka, Tomoichiro; Oseto, Mitsukai; Hedlund, Kjell-Olof

    2005-01-01

    Sapovirus, a member of the family Caliciviridae, is an etiologic agent of gastroenteritis in humans and pigs. Analyses of the complete genome sequences led us to identify the first sapovirus intergenogroup recombinant strain. Phylogenetic analysis of the nonstructural region (i.e., genome start to capsid start) grouped this strain into genogroup II, whereas the structural region (i.e., capsid start to genome end) grouped this strain into genogroup IV. We found that a recombination event occurred at the polymerase and capsid junction. This is the first report of intergenogroup recombination for any calicivirus and highlights a possible route of zoonoses because sapovirus strains that infect pig species belong to genogroup III. PMID:16485479

  19. Recombination experiments at CRYRING

    Energy Technology Data Exchange (ETDEWEB)

    Spies, W.; Glans, P.; Zong, W.; Gao, H.; Andler, G.; Justiniano, E.; Saito, M.; Schuch, R

    1998-11-15

    Recent advances in studies of electron-ion recombination processes at low relative energies with the electron cooler of the heavy-ion storage ring CRYRING are shown. Through the use of an adiabatically expanded electron beam, collisions down to 10{sup -4}eV relative energies were measured with highly charged ions stored in the ring at around 15 MeV/amu energies. Examples of recombination measurements for bare ions of D{sup +}, He{sup 2+}, N{sup 7+}, Ne{sup 10+} and Si{sup 14+} are presented. Further on, results of an experiment measuring laser-induced recombination (LIR) into n=3 states of deuterium with polarized laser light are shown.

  20. Recombinant Helicobacter pylori catalase

    Institute of Scientific and Technical Information of China (English)

    Yang Bai; Ya-Li Zhang; Jian-Feng Jin; Ji-De Wang; Zhao-Shan Zhang

    2003-01-01

    AIM: To construct a recombinant strain which highly expresses catalase of Helicobacter pylori(H.pylori) and assay the activity of H. pylori catalase.METHODS: The catalase DNA was amplified from H. pylori chromosomal DNA with PCR techniques and inserted into the prokaryotie expression vector pET-22b (+), and then was transformed into the BL21 (DE3) E. coli strain which expressed catalase recombinant protein. The activity of H.pylori catalase was assayed by the Beers & Sizers.RESULTS: DNA sequence analysis showed that the sequence of catalase DNA was the same as GenBank's research. The catalase recombinant protein amounted to 24.4 % of the total bacterial protein after induced with IPTG for 3 hours at 37 ℃ and the activity of H. pylori catalase was high in the BL21 (DE3) E. coli strain.CONCLUSION: A clone expressing high activity H. pylori catalase is obtained, laying a good foundation for further studies.

  1. Sex recombination, and reproductive fitness: an experimental study using Paramecium

    Energy Technology Data Exchange (ETDEWEB)

    Nyberg, D.

    1982-08-01

    The effect of sex and recombination on reproductive fitness are measured using five wild stocks of Paramecium primaurelia. Among the wild stocks there were highly significant differences in growth rates. No hybrid had as low a fitness as the least fit parental stock. Recombination produced genotypes of higher fitness than those of either parent only in the cross between the two stocks of lowest fitness. The increase in variance of fitness as a result of recombination was almost exclusively attributable to the generation lines with low fitness. The fitness consequences of sexuality and mate choice were stock specific; some individuals leaving the most descendants by inbreeding, others by outcrossing. For most crosses the short-term advantage of sex, if any, accrue from the fusion of different gametes (hybrid vigor) and not from recombination. Since the homozygous genotype with the highest fitnes left the most progeny by inbreeding (no recombination), the persistence of conjugation in P. primaurelia is paradoxical. (JMT)

  2. Insertion DNA Accelerates Meiotic Interchromosomal Recombination in Arabidopsis thaliana.

    Science.gov (United States)

    Sun, Xiao-Qin; Li, Ding-Hong; Xue, Jia-Yu; Yang, Si-Hai; Zhang, Yan-Mei; Li, Mi-Mi; Hang, Yue-Yu

    2016-08-01

    Nucleotide insertions/deletions are ubiquitous in eukaryotic genomes, and the resulting hemizygous (unpaired) DNA has significant, heritable effects on adjacent DNA. However, little is known about the genetic behavior of insertion DNA. Here, we describe a binary transgenic system to study the behavior of insertion DNA during meiosis. Transgenic Arabidopsis lines were generated to carry two different defective reporter genes on nonhomologous chromosomes, designated as "recipient" and "donor" lines. Double hemizygous plants (harboring unpaired DNA) were produced by crossing between the recipient and the donor, and double homozygous lines (harboring paired DNA) via self-pollination. The transfer of the donor's unmutated sequence to the recipient generated a functional β-glucuronidase gene, which could be visualized by histochemical staining and corroborated by polymerase chain reaction amplification and sequencing. More than 673 million seedlings were screened, and the results showed that meiotic ectopic recombination in the hemizygous lines occurred at a frequency  >6.49-fold higher than that in the homozygous lines. Gene conversion might have been exclusively or predominantly responsible for the gene correction events. The direct measurement of ectopic recombination events provided evidence that an insertion, in the absence of an allelic counterpart, could scan the entire genome for homologous counterparts with which to pair. Furthermore, the unpaired (hemizygous) architectures could accelerate ectopic recombination between itself and interchromosomal counterparts. We suggest that the ectopic recombination accelerated by hemizygous architectures may be a general mechanism for interchromosomal recombination through ubiquitously dispersed repeat sequences in plants, ultimately contributing to genetic renovation and eukaryotic evolution.

  3. Recombinant Protein Production and Insect Cell Culture and Process

    Science.gov (United States)

    Spaulding, Glenn F. (Inventor); Goodwin, Thomas J. (Inventor); OConnor, Kim C. (Inventor); Francis, Karen M. (Inventor); Andrews, Angela D. (Inventor); Prewett, Tracey L. (Inventor)

    1997-01-01

    A process has been developed for recombinant production of selected polypeptides using transformed insect cells cultured in a horizontally rotating culture vessel modulated to create low shear conditions. A metabolically transformed insect cell line is produced using the culture procedure regardless of genetic transformation. The recombinant polypeptide can be produced by an alternative process using virtually infected or stably transformed insect cells containing a gene encoding the described polypeptide. The insect cells can also be a host for viral production.

  4. Induced lead binding phytochelatins in Brassica juncea and Sesuvium portulacastrum investigated by orthogonal chromatography inductively coupled plasma-mass spectrometry and matrix assisted laser desorption ionization-time of flight-mass spectrometry.

    Science.gov (United States)

    Zaier, H; Mudarra, A; Kutscher, D; Fernández de la Campa, M R; Abdelly, C; Sanz-Medel, A

    2010-06-25

    The accumulation and transport of lead in Brassica juncea and Sesuvium portulacastrum plants and the possible formation of complexes of this element with bioligands such as phytochelatins was studied in roots and shoots of plants exposed to different amounts of Pb(NO(3))(2). Speciation studies on the plant extracts were conducted using size exclusion liquid chromatography and ion pair liquid chromatography coupled to UV and ICP-MS to monitor lead and sulphur. The identification of the species separated by chromatography was performed by MALDI-TOF-MS. In both types of exposed plants it was possible to identify the presence of the phytochelatin isoform PC(3). The results obtained suggest that both types of plants can be useful in studies of phytoremediation but the ability of S. portulacastrum to accumulate and redistribute Pb from root to shoot is more effective than B. juncea.

  5. Recombineering linear BACs.

    Science.gov (United States)

    Chen, Qingwen; Narayanan, Kumaran

    2015-01-01

    Recombineering is a powerful genetic engineering technique based on homologous recombination that can be used to accurately modify DNA independent of its sequence or size. One novel application of recombineering is the assembly of linear BACs in E. coli that can replicate autonomously as linear plasmids. A circular BAC is inserted with a short telomeric sequence from phage N15, which is subsequently cut and rejoined by the phage protelomerase enzyme to generate a linear BAC with terminal hairpin telomeres. Telomere-capped linear BACs are protected against exonuclease attack both in vitro and in vivo in E. coli cells and can replicate stably. Here we describe step-by-step protocols to linearize any BAC clone by recombineering, including inserting and screening for presence of the N15 telomeric sequence, linearizing BACs in vivo in E. coli, extracting linear BACs, and verifying the presence of hairpin telomere structures. Linear BACs may be useful for functional expression of genomic loci in cells, maintenance of linear viral genomes in their natural conformation, and for constructing innovative artificial chromosome structures for applications in mammalian and plant cells.

  6. Recombinant DNA for Teachers.

    Science.gov (United States)

    Duvall, James G., III

    1992-01-01

    A science teacher describes his experience at a workshop to learn to teach the Cold Spring Harbor DNA Science Laboratory Protocols. These protocols lead students through processes for taking E. coli cells and transforming them into a new antibiotic resistant strain. The workshop featured discussions of the role of DNA recombinant technology in…

  7. Recombinant renewable polyclonal antibodies.

    Science.gov (United States)

    Ferrara, Fortunato; D'Angelo, Sara; Gaiotto, Tiziano; Naranjo, Leslie; Tian, Hongzhao; Gräslund, Susanne; Dobrovetsky, Elena; Hraber, Peter; Lund-Johansen, Fridtjof; Saragozza, Silvia; Sblattero, Daniele; Kiss, Csaba; Bradbury, Andrew R M

    2015-01-01

    Only a small fraction of the antibodies in a traditional polyclonal antibody mixture recognize the target of interest, frequently resulting in undesirable polyreactivity. Here, we show that high-quality recombinant polyclonals, in which hundreds of different antibodies are all directed toward a target of interest, can be easily generated in vitro by combining phage and yeast display. We show that, unlike traditional polyclonals, which are limited resources, recombinant polyclonal antibodies can be amplified over one hundred million-fold without losing representation or functionality. Our protocol was tested on 9 different targets to demonstrate how the strategy allows the selective amplification of antibodies directed toward desirable target specific epitopes, such as those found in one protein but not a closely related one, and the elimination of antibodies recognizing common epitopes, without significant loss of diversity. These recombinant renewable polyclonal antibodies are usable in different assays, and can be generated in high throughput. This approach could potentially be used to develop highly specific recombinant renewable antibodies against all human gene products.

  8. A Recombinant Human Pluripotent Stem Cell Line Stably Expressing Halide-Sensitive YFP-I152L for GABAAR and GlyR-Targeted High-Throughput Drug Screening and Toxicity Testing

    Science.gov (United States)

    Kuenzel, Katharina; Friedrich, Oliver; Gilbert, Daniel F.

    2016-01-01

    GABAARs and GlyRs are considered attractive drug targets for therapeutic intervention and are also increasingly recognized in the context of in vitro neurotoxicity (NT) and developmental neurotoxicity (DNT) testing. However, systematic human-specific GABAAR and GlyR-targeted drug screening and toxicity testing is hampered due to lack of appropriate in vitro models that express native GABAARs and GlyRs. We have established a human pluripotent stem cell line (NT2) stably expressing YFP-I152L, a halide-sensitive variant of yellow fluorescent protein (YFP), allowing for fluorescence-based functional analysis of chloride channels. Upon stimulation with retinoic acid, NT2 cells undergo neuronal differentiation and allow pharmacological and toxicological evaluation of native GABAARs and GlyRs at different stages of brain maturation. We applied the cell line in concentration-response experiments with the neurotransmitters GABA and glycine as well as with the drugs strychnine, picrotoxin, fipronil, lindane, bicuculline, and zinc and demonstrate that the established in vitro model is applicable to GABAAR and GlyR-targeted pharmacological and toxicological profiling. We quantified the proportion of GABAAR and GlyR-sensitive cells, respectively, and identified percentages of approximately 20% each within the overall populations, rendering the cells a suitable model for systematic in vitro GABAAR and GlyR-targeted screening in the context of drug development and NT/DNT testing. PMID:27445687

  9. Effect of G gene-deleted recombinant viral hemorrhagic septicemia virus (rVHSV-ΔG) on the replication of wild type VHSV in a fish cell line and in olive flounder (Paralichthys olivaceus).

    Science.gov (United States)

    Kim, Min Sun; Choi, Seung Hyuk; Kim, Ki Hong

    2016-07-01

    In an earlier study, we generated a replicon viral hemorrhagic septicemia virus (VHSV) particle that was lacking the G gene in the genome (rVHSV-ΔG), and proved the potential of it as a protective vaccine through the immunization of olive flounder (Paralichthys olivaceus) fingerlings. Safety is the most important preconsideration for the development of recombinant live vaccines, and a major concern of propagation-incompetent viral particles would be the possible harmful effect to hosts through the interaction with wild-type viruses. Thus, in the present study, we analyzed the replication of rVHSV-ΔG in the presence of wild-type VHSV and the effect of rVHSV-ΔG on the replication of wild-type VHSV in Epithelioma papulosum cyprini (EPC) cells and in olive flounder fingerlings. The replication of wild-type VHSV in EPC cells was severely suppressed when the MOI of rVHSV-ΔG was 0.1 or 0.01, on the other hand, the titers of rVHSV-ΔG were not increased and stayed in a relatively constant according to time lapse. Furthermore, the replication of other novirhabdoviruses, IHNV and HIRRV, was also inhibited by co-infection with high titers of rVHSV-ΔG. There were no big differences in mortalities between groups infected with wild-type VHSV plus rVHSV-ΔG and groups infected with wild-type VHSV alone, when the challenged wild-type VHSV was more than 10(2) PFU/fish. However, a group of fish infected with 10 PFU/fish of wild-type VHSV plus rVHSV-ΔG showed significantly lower and slowly progressing cumulative mortality than a group of fish infected with 10 PFU/fish of wild-type VHSV alone. This result suggests that rVHSV-ΔG has an ability to attenuate the disease progression caused by wild-type VHSV when co-infected with relatively low titers of wild-type VHSV. These results indicate that the propagation-incompetent rVHSV-ΔG would not worsen but attenuate the progression of a disease caused by wild-type VHSV infection. Therefore, rVHSV-ΔG-based vaccines can provide a

  10. Algae-based oral recombinant vaccines.

    Science.gov (United States)

    Specht, Elizabeth A; Mayfield, Stephen P

    2014-01-01

    Recombinant subunit vaccines are some of the safest and most effective vaccines available, but their high cost and the requirement of advanced medical infrastructure for administration make them impractical for many developing world diseases. Plant-based vaccines have shifted that paradigm by paving the way for recombinant vaccine production at agricultural scale using an edible host. However, enthusiasm for "molecular pharming" in food crops has waned in the last decade due to difficulty in developing transgenic crop plants and concerns of contaminating the food supply. Microalgae could be poised to become the next candidate in recombinant subunit vaccine production, as they present several advantages over terrestrial crop plant-based platforms including scalable and contained growth, rapid transformation, easily obtained stable cell lines, and consistent transgene expression levels. Algae have been shown to accumulate and properly fold several vaccine antigens, and efforts are underway to create recombinant algal fusion proteins that can enhance antigenicity for effective orally delivered vaccines. These approaches have the potential to revolutionize the way subunit vaccines are made and delivered - from costly parenteral administration of purified protein, to an inexpensive oral algae tablet with effective mucosal and systemic immune reactivity.

  11. Algae-based oral recombinant vaccines

    Directory of Open Access Journals (Sweden)

    Elizabeth A Specht

    2014-02-01

    Full Text Available Recombinant subunit vaccines are some of the safest and most effective vaccines available, but their high cost and the requirement of advanced medical infrastructure for administration make them impractical for many developing world diseases. Plant-based vaccines have shifted that paradigm by paving the way for recombinant vaccine production at agricultural scale using an edible host. However, enthusiasm for molecular pharming in food crops has waned in the last decade due to difficulty in developing transgenic crop plants and concerns of contaminating the food supply. Microalgae are poised to become the next candidate in recombinant subunit vaccine production, and they present several advantages over terrestrial crop plant-based platforms including scalable and contained growth, rapid transformation, easily obtained stable cell lines, and consistent transgene expression levels. Algae have been shown to accumulate and properly fold several vaccine antigens, and efforts are underway to create recombinant algal fusion proteins that can enhance antigenicity for effective orally-delivered vaccines. These approaches have the potential to revolutionize the way subunit vaccines are made and delivered – from costly parenteral administration of purified protein, to an inexpensive oral algae tablet with effective mucosal and system immune reactivity.

  12. Variation in concentrations of high-molecular-weight glutenin subunits and macropolymers in wheat grains of a recombinant inbred lines population and in two contrasting eco-sites in China

    DEFF Research Database (Denmark)

    Li, Xiangnan; Cai, Jian; Liu, Fulai;

    2012-01-01

    BACKGROUND: Concentrations of high-molecular-weight glutenin subunits andmacropolymers in wheat grains are important indicators of grain quality, which are genetically determined and affected by environmental factors. The 6 VS·6AL translocation chromosome segment is reported to own high powdery...... in concentrations of HMW-GS and GMP were observed within lines containing different HMW-GS compositions and between the contrasting eco-sites. However, 6 VS·6AL chromosome translocation segment showed no significant effects on concentrations ofHMW-GSandGMP. In addition,HMW-GSconcentration was also found...... to be significantly correlated with the GMP concentration. CONCLUSION: Concentrations of HMW-GS andGMPare largely affected by the eco-sites and the composition ofHMW-GS, whilst not by the presence of 6 VS·6AL chromosome segment translocation. The 6 VS·6AL translocation is suggested as potential donor for breeding...

  13. 利用RNA干扰技术构建IER5-SiRNA-Hela 细胞系%Recombination IER5-siRNA-Hela cells line using RNAi technology

    Institute of Scientific and Technical Information of China (English)

    李莉; 赵焕英; 杨川杰; 鲁苇媛; 郝淳; 乔茶; 周平坤; 徐勤芝; 丁库克

    2011-01-01

    Objective To investigate the effect of RNA interference targeting IER5 gene on Hela cells in vitro and screen highly efficient small interfering RNA (siRNA ) .Methods RNAi technology was used to make the IER5 RNAi expression vector to transfect to Hela cells and established the stable transfected cell lines .Results Comparing the IER5-siRNA-Hela to the normal Hela cells ,the IER5 knock down Hela cells (IER5-siRNA-Hela) has high number as well as with a larger cell size using real-time PCR and Westem blot analysis .Conclusion We confinned the IER5 RNAi could effectively knock down the endogenous IER 5 from the Hela cells ,and established the stable transfected cell lines .%目的 观察小干扰RNA(siRNA)转染Hela细胞后,对IER5基因抑制效果,筛选出高效的siRNA转染质粒.方法 利用siRNA设计软件,将设计好的RNA片断插入到siRNA表达质粒载体PsilencerTM 3.1-H1 hygro中,测序正确后转染至Hela细胞中.利用Real-time PCR的方法,测定siRNA对Hela细胞中IRE5的抑制效果.结果 在mRNA水平及Western blot测试,IER5-siRNA-Hela细胞的IER5基因mRNA表达抑制效果明显.从细胞形态上讲,IER5-siRNA-Hela细胞与正常Hela细胞相比,IER5-siRNA-Hela细胞比Hela细胞体积大.结论 本实验成功的获得了特异性抑制IER5基因的IER5-siRNA-Hela细胞系.

  14. SUMO Wrestles with Recombination

    Directory of Open Access Journals (Sweden)

    Lumír Krejčí

    2012-07-01

    Full Text Available DNA double-strand breaks (DSBs comprise one of the most toxic DNA lesions, as the failure to repair a single DSB has detrimental consequences on the cell. Homologous recombination (HR constitutes an error-free repair pathway for the repair of DSBs. On the other hand, when uncontrolled, HR can lead to genome rearrangements and needs to be tightly regulated. In recent years, several proteins involved in different steps of HR have been shown to undergo modification by small ubiquitin-like modifier (SUMO peptide and it has been suggested that deficient sumoylation impairs the progression of HR. This review addresses specific effects of sumoylation on the properties of various HR proteins and describes its importance for the homeostasis of DNA repetitive sequences. The article further illustrates the role of sumoylation in meiotic recombination and the interplay between SUMO and other post-translational modifications.

  15. Recombinant Human Enterovirus 71

    OpenAIRE

    2004-01-01

    Two human enterovirus 71 (HEV71) isolates were identified from hand, foot and mouth disease patients with genome sequences that had high similarity to HEV71 (>93%) at 5´ UTR, P1, and P2 and coxsackievirus A16 (CV-A16, >85%) at P3 and 3´UTR. Intertypic recombination is likely to have occurred between HEV71 and CV-A16 or an as-yet to be described CV-A16-like virus.

  16. Recombinant human milk proteins.

    Science.gov (United States)

    Lönnerdal, Bo

    2006-01-01

    Human milk provides proteins that benefit newborn infants. They not only provide amino acids, but also facilitate the absorption of nutrients, stimulate growth and development of the intestine, modulate immune function, and aid in the digestion of other nutrients. Breastfed infants have a lower prevalence of infections than formula-fed infants. Since many women in industrialized countries choose not to breastfeed, and an increasing proportion of women in developing countries are advised not to breastfeed because of the risk of HIV transmission, incorporation of recombinant human milk proteins into infant foods is likely to be beneficial. We are expressing human milk proteins known to have anti-infective activity in rice. Since rice is a normal constituent of the diet of infants and children, limited purification of the proteins is required. Lactoferrin has antimicrobial and iron-binding activities. Lysozyme is an enzyme that is bactericidal and also acts synergistically with lactoferrin. These recombinant proteins have biological activities identical to their native counterparts. They are equally resistant to heat processing, which is necessary for food applications, and to acid and proteolytic enzymes which are needed to maintain their biological activity in the gastrointestinal tract of infants. These recombinant human milk proteins may be incorporated into infant formulas, baby foods and complementary foods, and used with the goal to reduce infectious diseases.

  17. Produção e aspectos econômicos de milho consorciado com Crotalária juncea L. em diferentes intervalos de semeadura, sob manejo orgânico

    Directory of Open Access Journals (Sweden)

    Emerson Dalla Chieza

    Full Text Available RESUMO A busca por sistemas produtivos mais sustentáveis é um desafio necessário. Avaliaram-se formas de manejo do consórcio entre milho e Crotalaria juncea L., que possibilitem otimizar a produção “in situ” de massa vegetal, sem comprometer o potencial produtivo do cereal. Realizaram-se experimentos em épocas distintas: primavera-verão (experimento 1 e outono-inverno (experimento 2, em Seropédica -RJ, no período de 11/2010 a 09/2011. Os tratamentos para o experimento 1 foram: C. juncea, semeada: sete dias antes, simultaneamente com, aos 14 e aos 28 dias após a cultura do milho, respectivamente, para T1, T2, T3 e T4. Além desses, T5-milho em monocultivo e T6-milho em monocultivo + N. Os tratamentos para o experimento 2 foram: C. juncea semeada: sete dias antes, simultaneamente com, aos sete e aos 14 dias após a cultura do milho, respectivamente para TA, TB, TC e TD. Além desses, TE-milho em monocultivo e TF-milho em monocultivo + N. Para T1, T2, TA e TB a C. juncea foi roçada aproximadamente 30 dias após a emergência e sua palhada utilizada como fonte de N para o milho; já para T3, T4, T6, TC, TD e TF foi utilizada a torta de mamona como fonte de N. Na primavera-verão, em T2, a produção de grãos do cereal não diferiu da produção do monocultivo com adubação nitrogenada, com reflexo positivo no balanço econômico. No outono-inverno, a presença do adubo verde semeado 14 dias após o milho resultou em benefícios ao sistema. A C. juncea, quando cultivada na primavera-verão, apresentou potencial para fornecer N ao milho como alternativa à adubação nitrogenada de cobertura.

  18. Comparative mapping of Brassica juncea and Arabidopsis thaliana using Intron Polymorphism (IP markers: homoeologous relationships, diversification and evolution of the A, B and C Brassica genomes

    Directory of Open Access Journals (Sweden)

    Gupta Vibha

    2008-03-01

    Full Text Available Abstract Background Extensive mapping efforts are currently underway for the establishment of comparative genomics between the model plant, Arabidopsis thaliana and various Brassica species. Most of these studies have deployed RFLP markers, the use of which is a laborious and time-consuming process. We therefore tested the efficacy of PCR-based Intron Polymorphism (IP markers to analyze genome-wide synteny between the oilseed crop, Brassica juncea (AABB genome and A. thaliana and analyzed the arrangement of 24 (previously described genomic block segments in the A, B and C Brassica genomes to study the evolutionary events contributing to karyotype variations in the three diploid Brassica genomes. Results IP markers were highly efficient and generated easily discernable polymorphisms on agarose gels. Comparative analysis of the segmental organization of the A and B genomes of B. juncea (present study with the A and B genomes of B. napus and B. nigra respectively (described earlier, revealed a high degree of colinearity suggesting minimal macro-level changes after polyploidization. The ancestral block arrangements that remained unaltered during evolution and the karyotype rearrangements that originated in the Oleracea lineage after its divergence from Rapa lineage were identified. Genomic rearrangements leading to the gain or loss of one chromosome each between the A-B and A-C lineages were deciphered. Complete homoeology in terms of block organization was found between three linkage groups (LG each for the A-B and A-C genomes. Based on the homoeology shared between the A, B and C genomes, a new nomenclature for the B genome LGs was assigned to establish uniformity in the international Brassica LG nomenclature code. Conclusion IP markers were highly effective in generating comparative relationships between Arabidopsis and various Brassica species. Comparative genomics between the three Brassica lineages established the major rearrangements

  19. A Glance at Recombination Hotspots in the Domestic Cat.

    Directory of Open Access Journals (Sweden)

    Hasan Alhaddad

    Full Text Available Recombination has essential roles in increasing genetic variability within a population and in ensuring successful meiotic events. The objective of this study is to (i infer the population-scaled recombination rate (ρ, and (ii identify and characterize regions of increased recombination rate for the domestic cat, Felis silvestris catus. SNPs (n = 701 were genotyped in twenty-two East Asian feral cats (random bred. The SNPs covered ten different chromosomal regions (A1, A2, B3, C2, D1, D2, D4, E2, F2, X with an average region size of 850 Kb and an average SNP density of 70 SNPs/region. The Bayesian method in the program inferRho was used to infer regional population recombination rates and hotspots localities. The regions exhibited variable population recombination rates and four decisive recombination hotspots were identified on cat chromosome A2, D1, and E2 regions. As a description of the identified hotspots, no correlation was detected between the GC content and the locality of recombination spots, and the hotspots enclosed L2 LINE elements and MIR and tRNA-Lys SINE elements.

  20. Bacterial Recombineering: Genome Engineering via Phage-Based Homologous Recombination.

    Science.gov (United States)

    Pines, Gur; Freed, Emily F; Winkler, James D; Gill, Ryan T

    2015-11-20

    The ability to specifically modify bacterial genomes in a precise and efficient manner is highly desired in various fields, ranging from molecular genetics to metabolic engineering and synthetic biology. Much has changed from the initial realization that phage-derived genes may be employed for such tasks to today, where recombineering enables complex genetic edits within a genome or a population. Here, we review the major developments leading to recombineering becoming the method of choice for in situ bacterial genome editing while highlighting the various applications of recombineering in pushing the boundaries of synthetic biology. We also present the current understanding of the mechanism of recombineering. Finally, we discuss in detail issues surrounding recombineering efficiency and future directions for recombineering-based genome editing.

  1. Sex influence on recombination frequency in Secale cereale L.

    Science.gov (United States)

    Benito, C; Romero, M P; Henriques-Gil, N; Llorente, F; Figueiras, A M

    1996-10-01

    The variation in recombination frequency (rf) is important to plant breeders since their major objective is to obtain favorable recombinants of linked genes. One source of variation in rf is sex. Sex differences for recombination frequencies were studied in four of the seven chromosomes of Secale cereale L. cv 'Ailés' using isozyme and storage protein loci and were determined on the basis of reciprocal crosses between heterozygous plants of cv. 'Ailés' and homozygous plants of the inbred line 'Riodeva'. The differences were found to be strongly segmentspecific. In some cases the level of crossing-over in male and female meiosis was about the same (between Pgm1 and Ndh1 loci on chromosome arm 4RS). However, for most of the chromosome segments in 1R, 3RL and 6RL the male rf was significantly higher than the female rf. Different hypotheses about the mechanisms of plant sex differences for recombination are discussed.

  2. Recombinant Collagenlike Proteins

    Science.gov (United States)

    Fertala, Andzej

    2007-01-01

    A group of collagenlike recombinant proteins containing high densities of biologically active sites has been invented. The method used to express these proteins is similar to a method of expressing recombinant procollagens and collagens described in U. S. Patent 5,593,859, "Synthesis of human procollagens and collagens in recombinant DNA systems." Customized collagenous proteins are needed for biomedical applications. In particular, fibrillar collagens are attractive for production of matrices needed for tissue engineering and drug delivery. Prior to this invention, there was no way of producing customized collagenous proteins for these and other applications. Heretofore, collagenous proteins have been produced by use of such biological systems as yeasts, bacteria, and transgenic animals and plants. These products are normal collagens that can also be extracted from such sources as tendons, bones, and hides. These products cannot be made to consist only of biologically active, specific amino acid sequences that may be needed for specific applications. Prior to this invention, it had been established that fibrillar collagens consist of domains that are responsible for such processes as interaction with cells, binding of growth factors, and interaction with a number of structural proteins present in the extracellular matrix. A normal collagen consists of a sequence of domains that can be represented by a corresponding sequence of labels, e.g., D1D2D3D4. A collagenlike protein of the present invention contains regions of collagen II that contain multiples of a single domain (e.g., D1D1D1D1 or D4D4D4D4) chosen for its specific biological activity. By virtue of the multiplicity of the chosen domain, the density of sites having that specific biological activity is greater than it is in a normal collagen. A collagenlike protein according to this invention can thus be made to have properties that are necessary for tissue engineering.

  3. Dielectronic recombination theory

    Energy Technology Data Exchange (ETDEWEB)

    LaGattuta, K.J.

    1991-12-31

    A theory now in wide use for the calculation of dielectronic recombination cross sections ({sigma}{sup DR}) and rate coefficients ({alpha}{sup DR}) was one introduced originally by Feshbach for nuclear physics applications, and then later adapted for atomic scattering problems by Hahn. In the following, we briefly review this theory in a very general form, which allows one to account for the effects of overlapping and interacting resonances, as well as continuum-continuum coupling. An extension of our notation will then also allow for the inclusion of the effects of direct radiative recombination, along with a treatment of the interference between radiative and dielectronic recombination. Other approaches to the calculation of {sigma}{sup DR} have been described by Fano and by Seaton. We will not consider those theories here. Calculations of {alpha}{sup DR} have progressed considerably over the last 25 years, since the early work of Burgess. Advances in the reliability of theoretical predictions have also been promoted recently b a variety of direct laboratory measurements of {sigma}{sup DR}. While the measurements of {sigma}{sup DR} for {delta}n {ne} 0 excitations have tended to agree very well with calculations, the case of {delta}n = 0 has been much problematic. However, by invoking a mechanism originally proposed by Jacobs, which takes into account the effect of stray electric fields on high Rydberg states (HRS) participating in the DR process, new calculations have improved the agreement between theory and experiment for these cases. Nevertheless, certain discrepancies still remain.

  4. Did the universe recombine

    Energy Technology Data Exchange (ETDEWEB)

    Bartlett, J.G.; Stebbins, A. (California, University, Berkeley (USA) Toronto, University (Canada))

    1991-04-01

    The Zel'dovich-Sunyaev model-independent arguments for the existence of a neutral hydrogen phase is reviewed in light of new limits on the Compton y parameter from COBE. It is concluded that with baryon densities compatible with standard cosmological nucleosynthesis, the universe could have remained fully ionized throughout its history without producing a detectable spectral distortion. It is argued that it is unlikely that spectral observations of the cosmic microwave background will ever require the universe to have recombined for flat cosmologies. 22 refs.

  5. 川麦42/川农16重组自交系群体中小麦迟α-淀粉酶(LMA)的遗传分析%Inheritance of late maturityα-amylase(LMA) in a recombined inbreed lines of Chuanmai 42 crossed with Chuannong 16

    Institute of Scientific and Technical Information of China (English)

    杨恩年; 张颙; 李俊; 汤永禄; 杨武云; 邹裕春

    2007-01-01

    利用川麦42/川农16的重组自交系(recombined inbreed lines,RILs)群体,采用双抗体夹心酶联免疫吸附(ELISA)试剂盒,对迟熟α-淀粉酶(Late maturityα-amylase,LMA)进行了测试;选用7B染色体上的3对与LMA相关的SSRs引物(Xgwm577,Xwmc276,Xwmc273),对群体中标记与LMA的相关性进行了验证;同时对LMA和降落值(Falling Number,FN)的相关性进行了分析.结果表明:①LMA是受一对基因控制;②3对SSRs引物(Xgwm577,Xwmc276,Xwmc273)在MILs中对LMA的检测没有作用.③在本试验的MILs中,LMA的存在对降落值又重要影响,同时降落值还受其它遗传控制.

  6. The Richness and Beauty of the Physics of Cosmological Recombination

    Science.gov (United States)

    Sunyaev, Rashid

    2009-01-01

    In our Universe the initial temperature of radiation was very high and hydrogen and helium were completely ionized. At redshifts z 1400 the temperature dropped to 3800 K and, according to the Saha equation, the recombination of hydrogen should occur. In reality this process is strongly delayed and some frozen amount of electrons should be present till the reionization of the Universe at z 10. Process of recombination defines the position and the width of the last scattering surface which is crucial for the formation of the observed angular fluctuations of cosmic microwave background radiation (CMB), acoustic peaks and barionic oscillations in the distribution of galaxies and clusters of galaxies in space. The recombination of hydrogen occurs under conditions of very low density and in the presence of black body radiation. As a result, usually insignificant atomic processes begin to play a role. They influence the shape of acoustic peaks at a level which will be detectable by the Planck Surveyor spacecraft and we should take them into account when estimating the key parameters of the Universe from CMB data. The recombination of hydrogen and helium leads to the appearance of recombinational lines in centimeter and decimeter spectral bands. Observations of these lines will make it possible to check the predictions of the big bang recombination theory and will open a possibility to measure directly the density of barions, the CMB monopole temperature and specific entropy of the Universe. Observations of helium recombination lines originated at redshifts 6000 and 2500 will open a way to measure the prestellar abundance of helium in the Universe.

  7. Novel recombinant sapovirus in Bangladesh.

    Science.gov (United States)

    Dey, Shuvra Kanti; Mizuguchi, Masashi; Okitsu, Shoko; Ushijima, Hiroshi

    2011-01-01

    Recombination of RNA viruses plays an important part in molecular epidemiological study, virus evolution, vaccine design, and viral control programs. Sapovirus, a member of the family Caliciviridae, is one of the major causative agents of viral gastroenteritis affecting all age groups. Sapovirus capsid and polymerase regions were amplified by PCR using specific primers. PCR products were sequenced directly and sequence analysis was performed using CLUSTAL X, SimPlot, and MEGA 4 software package. Based on the genetic analysis, a novel, naturally occurring recombinant sapovirus strain was identified in Bangladesh. Breakpoint analysis of the recombinant sapovirus showed that the recombination site was at the open reading frame ORF1/ORF2 overlap. We described the genetic characterization of a novel, naturally occurring recombinant sapovirus and provided the first evidence of recombination in sapovirus in Bangladesh.

  8. Cell biology of mitotic recombination

    DEFF Research Database (Denmark)

    Lisby, Michael; Rothstein, Rodney

    2015-01-01

    Homologous recombination provides high-fidelity DNA repair throughout all domains of life. Live cell fluorescence microscopy offers the opportunity to image individual recombination events in real time providing insight into the in vivo biochemistry of the involved proteins and DNA molecules...... of this review include the stoichiometry and dynamics of recombination complexes in vivo, the choreography of assembly and disassembly of recombination proteins at sites of DNA damage, the mobilization of damaged DNA during homology search, and the functional compartmentalization of the nucleus with respect...... as well as the cellular organization of the process of homologous recombination. Herein we review the cell biological aspects of mitotic homologous recombination with a focus on Saccharomyces cerevisiae and mammalian cells, but will also draw on findings from other experimental systems. Key topics...

  9. Initiation of V(D)J recombination in a cell-free system

    NARCIS (Netherlands)

    D.C. van Gent (Dik); J.F. McBlane; D.A. Ramsden; M.J. Sadokfsky; J.E. Hesse (Joanne); M. Gellert

    1995-01-01

    textabstractCells performing V(D)J recombination make specific cuts in DNA at recombination signal sequences. Here, we show that nuclear extracts of pre-B cell lines carry out this specific cleavage. The products of cleavage are the same as found previously in thymocytes: full-leng

  10. Utilizing Protein-lean Co-products from Corn Containing Recombinant Pharmaceutical Proteins for Ethanol Production

    Science.gov (United States)

    Protein-lean fractions of corn (maize) containing recombinant (r) pharmaceutical proteins were used to produce fuel ethanol and residual r-proteins in the co-product, distillers dry grains with solubles (DDGS), were determined. Transgenic corn lines containing recombinant green fluorescence protein ...

  11. Identification of fungus-responsive cis-acting element in the promoter of Brassica juncea chitinase gene, BjCHI1.

    Science.gov (United States)

    Gao, Ying; Zan, Xin-Li; Wu, Xue-Feng; Yao, Lei; Chen, Yu-Ling; Jia, Shuang-Wei; Zhao, Kai-Jun

    2014-02-01

    Chitinases are a group of pathogenesis-related proteins. The Brassica juncea chitinase gene BjCHI1 is highly inducible by pathogenic fungal infection, suggesting that the promoter of BjCHI1 might contain specific cis-acting element responsive to fungal attack. To identify the fungus-responsive element in BjCHI1 promoter (BjC-P), a series of binary plant transformation vectors were constructed by fusing the BjC-P or its deletion-derivatives to β-glucuronidase (GUS) reporter gene. Expression of the GUS reporter gene was systematically assayed by a transient gene expression system in Nicotiana benthamiana leaves treated with fungal elicitor Hexa-N-Acetyl-Chitohexaose, as well as in transgenic Arabidopsis plants inoculated with fungus Botrytis cinerea. The histochemical and quantitative GUS assays showed that the W-box-like element (GTAGTGACTCAT) in the region (-668 to -657) was necessary for the fungus-response, although there were another five W-box-like elements in BjC-P. In addition, gain-of-function analysis demonstrated that the fragment (-409 to -337) coupled to the W-box-like element was needed for full magnitude of the fungal induction. These results revealed the existence of a novel regulation mechanism of W-box-like element involved in plant pathogenic resistance, and will benefit the potential application of BjC-P in engineering crops.

  12. Study on the mitochondrial activity and membrane potential after exposing later stage oocytes of two gorgonian corals (Junceella juncea and Junceella fragilis) to cryoprotectants.

    Science.gov (United States)

    Tsai, S; Spikings, E; Huang, I-C; Lin, C

    2011-01-01

    Coral reefs provide a valuable habitat for many economically valuable fish and invertebrates. However, they are in serious jeopardy, threatened by increasing over-exploitation, pollution, habitat destruction, disease and global climate change. Here, we examined the effect of cryoprotectant exposure on mitochondrial activity and membrane potential in coral oocytes in order to find suitable cryoprotectants towards their successful cryopreservation. According to the No Observed Effect Concentrations (NOECs), methanol was found to be the least toxic cryoprotectant whilst DMSO was the most toxic cryoprotectant. The results also demonstrated that there were no significant differences (p > 0.05) in ATP concentrations between Junceella juncea and Junceella fragilis after exposure to all concentrations of all cryoprotectants for 30 min. Using confocal microscopy, JC-1 (5,50,6,60-tetrachloro-1,10,3,30-tetraethyl-imidacarbocyanine iodide) staining indicated that the mitochondrial membrane potential of Junceella fragilis oocytes reduced after 1 M and 2 M methanol treatment and a loss of the mitochondrial distribution pattern and poor green fluorescence after 3M methanol treatment. Therefore, even oocytes that show no adverse effect of cryoprotectants on survival might suffer some more subtle impacts. The results obtained from this study will provide a basis for development of protocols to cryopreserve the oocytes of gorgonian corals.

  13. Significance of Bacillus subtilis strain SJ-101 as a bioinoculant for concurrent plant growth promotion and nickel accumulation in Brassica juncea.

    Science.gov (United States)

    Zaidi, Sabina; Usmani, Saima; Singh, Braj Raj; Musarrat, Javed

    2006-08-01

    In this study, a nickel (Ni)-tolerant Bacillus subtilis strain SJ-101 was characterized based on the 16SrDNA homology and phylogenetic analysis. The role of this strain ascertained in facilitating Ni accumulation in the Indian mustard plant (Brassica juncea [L]. Czern and Coss) var. Pusa Bold (DIR-50), to elucidate the potential of Ni phytoremediation in combination with metal-tolerant rhizobacteria. The data revealed that the plants exposed to NiCl2 (1750 mg kg(-1)) in soil bioaugmented with strain SJ-101 have accumulated 0.147% Ni vis-à-vis 0.094% accumulation in dry biomass of the plants grown in uninoculated soil. The strain SJ-101 has also exhibited the capability of producing indole acetic acid (IAA) (55 microg ml(-1)), and solubilizing inorganic phosphate (90 microg ml(-1)) in specific culture media. The pot culture experiments clearly demonstrated the beneficial effects of bioinoculant strain SJ-101 with significant increase (pplant growth attributes in untreated control soil. Furthermore, the protective effect of the strain SJ-101 against Ni phytotoxicity was evident in plants grown in soil treated with NiCl2 in concentration range of 250-1750 mg kg(-1). Thus, it is suggested that the strain SJ-101 owing to its intrinsic abilities of plant growth promotion, and attenuation of soil Ni by biosorption and bioaccumulation, could be exploited for bacteria-assisted phytoaccumulation of this toxic heavy metal from contaminated sites.

  14. Dissociative recombination of HCl+

    Science.gov (United States)

    Larson, Åsa; Fonseca dos Santos, Samantha; E. Orel, Ann

    2017-08-01

    The dissociative recombination of HCl+, including both the direct and indirect mechanisms, is studied. For the direct process, the relevant electronic states are calculated ab initio by combining electron scattering calculations to obtain resonance positions and autoionization widths with multi-reference configuration interaction calculations of the ion and Rydberg states. The cross section for the direct dissociation along electronic resonant states is computed by solution of the time-dependent Schrödinger equation. For the indirect process, an upper bound value for the cross section is obtained using a vibrational frame transformation of the elements of the scattering matrix at energies just above the ionization threshold. Vibrational excitations of the ionic core from the ground vibrational state, v = 0 , to the first three excited vibrational states, v = 1 , v = 2 , and v = 3 , are considered. Autoionization is neglected and the effect of the spin-orbit splitting of the ionic potential energy upon the indirect dissociative recombination cross section is considered. The calculated cross sections are compared to measurements.

  15. 14-3-3σ干扰逆转录病毒载体的构建及其稳定转染HaCat细胞系的建立%Construction of RNAi Recombinant Retroviral Vector of 14-3-3P and Its Stably Transfected HaCat Cell Lines

    Institute of Scientific and Technical Information of China (English)

    周美娟; 丁振华

    2011-01-01

    目的:构建14-3-3σ干扰逆转录病毒载体,建立稳定转染的HaCat细胞系.方法:人工合成14-3-3σ基因干扰序列并定向插入到pSuper-retro-neo-EGFP质粒,并在STBL3菌内进行质粒扩增,刷选阳性克隆,酶切测序鉴定,转染293FT细胞进行病毒包装、扩增、纯化、获取逆转录病毒载体,将逆转录病毒栽体感染HaCat细胞后Western免疫印迹法、Real-time PCR法检测14-3-3σ的表达情况.结果:连接重组后经酶切和测序筛选出pSuper-retro-neo-EGFP-si14-3-3σ;干扰质粒稳定转染的HaCat细胞系在倒置荧光显微镜下呈绿色荧光,Western免疫印迹法和Real-time PCR法表明14-3-3σ表达明显抑制.结论:成功构建了14-3-3σ干扰的逆转录病毒载体,并构建了其稳定转染的HaCat细胞系.%Objective: To construct the RNAi retroviral vector of 14-3-3σ and establish the stable transfected HaCat cell lines.Methods: Hairpin siRNA of 14-3-3σ was synthesized and inserted into pSuper-retro-neo-EGFP plasmid.PSuper-retro-neo-EGFP-si14-3-3σ was transformed into competent STBL3 cells.Then the positive clones were confirmed by sequencing and transfected into the packaging 293FT cells to amplificate and depurate virus.HaCat cells were infected by the recombinant retroviral vector and the expression of 14-3-3σ was detected by Western blot and real time PCR.Results: The recombinant retroviral plasmid PSuper-retro-neo-EGFP-si 14-3-3σ was successfully constructed and green fluorescence of the stable transfected HaCat cell lines were observed under inverted fluorescence microscope.The expression of 14-3-3 was down-regulated by the RNAi-14-3-3σ.Conclusion: The RNAi retroviral vector targeting 14-3-3σ was successfully constructed and stably transfected HaCat cell lines were established.

  16. Recombination-cascade X-ray spectra of highly charged helium-like ions

    Science.gov (United States)

    Pradhan, A. K.

    1985-01-01

    It is shown that the relative intensity distribution among the X-ray spectral lines of helium-like ions from the n = 2 states produced through recombination processes such as radiative and charge transfer recombination may be given by considering in detail the radiative cascades following recombination. Model calculations are presented with predicted line ratios for Ar XVII and Fe XXV in recombination-dominated noncoronal plasmas. In particular, compared to coronal intensities, the singlet resonance line (w) should be much weaker relative to the triplet intercombination (x, y) and forbidden (z) lines, yielding large values for the ratio G = (x + y + z)/w. Accurate configuration interaction type wave functions are employed to calculate the eigenenergies, transition probabilities, and cascade coefficients. Certain relevant tokamak and astrophysical observations are discussed.

  17. Visualization of site-specific recombination catalyzed by a recombinase from Zygosaccharomyces rouxii in Arabidopsis thaliana.

    Science.gov (United States)

    Onouchi, H; Nishihama, R; Kudo, M; Machida, Y; Machida, C

    1995-06-25

    Excision of a DNA segment can occur in Arabidopsis thaliana by reciprocal recombination between two specific recombination sites (RSs) when the recombinase gene (R) from Zygosaccharomyces rouxii is expressed in the plant. To monitor recombination events, we generated several lines of transgenic Arabidopsis plants that carried a cryptic beta-glucuronidase (GUS) reporter gene which was designed in such a way that expression of the reporter gene could be induced by R gene-mediated recombination. We also made several transgenic lines with an R gene linked to the 35S promoter of cauliflower mosaic virus. Each transgenic line carrying the cryptic reporter gene was crossed with each line carrying the R gene. Activity of GUS in F1 and F2 progeny was examined histochemically and recombination between two RSs was analyzed by Southern blotting and the polymerase chain reaction. In seedlings and plantlets of F1 progeny and most of the F2 progeny, a variety of patterns of activity of GUS, including sectorial chimerism in leaves, was observed. A small percentage of F2 individuals exhibited GUS activity in the entire plant. This pattern of expression was ascribed to germinal recombination in the F1 generation on the basis of an analysis of DNA structure by Southern blotting. These results indicate that R gene-mediated recombination can be induced in both somatic and germ cells of A. thaliana by cross-pollination of parental transgenic lines.

  18. Desempenho agronômico de linhas endogâmicas ecombinadas de Capsicum annuum L. em sistema orgânico sob cultivo protegido Agronomic performance of recombinant inbred of Capsicum annuum L. lines cultivated under organic system and greenhouse conditions

    Directory of Open Access Journals (Sweden)

    Sarah Ola Moreira

    2010-08-01

    of this work was to study the agronomic performance of 12 recombinant inbred chili pepper (Capsicum annuum L. lines, from crosses between UENF 1421 and UENF 1381 accessions, under greenhouse conditions and using an organic system. The experiment was carried out at Seropédica Experimental Station of PESAGRO-RIO, in randomized block design, with four replications. The following variables were evaluated: total number of fruits (TNF; total fruit weight (TFW; average weight of fruits (MWF; fruit length (FL; fruit diameter (FD; length and diameter fruit ratio (LDFR, and capsaicin presence (CAPS. Genotypic, phenotypic and environmental variances, as well as genotypic determination coefficient and variation index were estimated. There was highly significant difference for all traits unless TFW. The lines produced a high number of fruits, with low weight and with variability in shape among lines. Absence of capsaicin was observed only in two lines and other three lines segregated for capsaicin presence. High values of genotypic variance and genotypic determination coefficient were estimated for all traits, indicating that the results were due to genetic causes. The data pointed to a preliminary indication of lines 5 and 8 for cultivation in an organic system under greenhouse conditions.

  19. RECOMBINANT HORSERADISH PEROXIDASE FOR ANALYTICAL APPLICATIONS

    OpenAIRE

    2013-01-01

    The article deals with prospects of using recombinant horseradish peroxidase in analytical biochemistry and biotechnology. Problems of recombinant horseradish peroxidase cloning in different expression systems, possible approaches to their solution, advantages of recombinant recombinant horseradish peroxidase and recombinant horseradish peroxidase-fusion proteins for immunoassays are considered. Possibility for development of mediatorless bienzyme biosensor for peroxide and metabolites, yield...

  20. STUDI PAPARAN MEDAN MAGNET SALURAN UDARA TEGANGAN EKSTRA TINGGI (SUTET PADA PERTUMBUHAN SAYURAN CAISIM (BRASSICA JUNCEA L

    Directory of Open Access Journals (Sweden)

    I Gede Ketut Sri Budarsa

    2012-11-01

    Full Text Available The existence of 500 kv extra high voltage transmission lines causes some area of villager’s farms to be under the transmission lines, so the research is urgently needed to see how the exposure of magnetic fields of 500 kv extra high voltage transmission lines influences the growth of plants. The aim of this research main function is to know the growth response of green mustard with the exposure to the extra high voltage magnetic field. Observation is focused on the area of leaf, amount of chlorophyll, weight of dry leaf, and net assimilation rate .The research applies completely randomized design repeated 3 times and 4 kinds of treatments with the different exposure of magnetic field. The data are quantitatively analyzed using analysis of variant method. The result of this research shows that the exposure of magnetic field influences the reduction of green mustard leaf, amount of leaf chlorophyll, net assimilation rate and the dry weight of green mustard leaf. Based on the result of the research it can be concluded that the exposure of magnetic field can reduce the growth of green mustard witch are under 500 kv extra high voltage transmission lines

  1. On-line methanol sensor system development for recombinant ...

    African Journals Online (AJOL)

    PANCHIGA

    2016-10-19

    Oct 19, 2016 ... 2Pilot Plant Development and Training Institute (PDTI), 49 Soi Tientalay 25, Bangkhuntien-Chaithalay Road, .... three-stage fermentation process (Çelik and Çalik, 2012; ..... The authors thank Dr. Witoon Tirasophon of Shrimp.

  2. Radio Recombination Lines at Decametre Wavelengths. Prospects for the Future

    Science.gov (United States)

    2010-09-15

    quantum number levels are a sensi- tive probe of the environments in which the atoms exist, provid- ing diagnostics such as temperature, density, and...measured simultaneously at different quantum numbers n in the same spectrum, can be used to constrain the physical prop- erties of the absorbing gas...25 Years of Investigation, 163, 189 Stepkin, S. V., Konovalenko, A. A., Kantharia, N. G., & Udaya Shankar , N. 2007, MNRAS, 374, 852 6 W. M. Peters et

  3. The o II recombination line abundance problem in planetary nebulae

    Directory of Open Access Journals (Sweden)

    D. R. Garnett

    2001-01-01

    Full Text Available Presentamos observaciones de l neas de recombinaci on de O II en diez nebu- losas planetarias brillantes, y mediciones de O II y [O III] con resoluci on espacial en NGC 6720. Esto es con el n de estudiar la discrepancia entre las abundancias calculadas con l neas de recombinaci on de O II y aquellas derivadas de l neas de excitaci on colisional de [O III]. Observamos una gran variaci on en la discrepan- cia, desde tener resultados consistentes hasta una diferencia de un factor de seis. La magnitud de esta discrepancia esta anti-correlacionada con el brillo super cial de la nebulosa (aquellas que son compactas y m as brillantes tienen las menores variaciones. Los niveles de O II poblados a trav es de recombinaci on di-electr onica dan abundancias mayores que otros niveles. Nuestra observaci on en rendija larga muestra que la emisi on en O II tiene su m aximo al interior de la c ascara brillante donde [O III] y H son m as brillantes. Con base en las correlaciones observadas, proponemos que la intensa emisi on en l neas de recombinaci on en nebulosas plan- etarias proviene de recombinaci on di-electr onica aumentada en el interior de la nebulosa, la cual posiblemente est e impulsada por un viento estelar caliente.

  4. Collision and recombination driven instabilities in variable charged dusty plasmas

    Indian Academy of Sciences (India)

    S Bal; M Bose

    2013-04-01

    The dust-acoustic instability driven by recombination of electrons and ions on the surface of charged and variably-charged dust grains as well as by collisions in dusty plasmas with significant pressure of background neutrals have been theoretically investigated. The recombination driven instability is shown to be dominant in the long wavelength regime even in the presence of dust-neutral and ion-neutral collisions, while in the shorter wavelength regime, the dust-neutral collision is found to play a major role. In an earlier research work, the dust-neutral collision was neglected in comparison to the effect due to the recombination for estimating the dust-acoustic instability; later the other report shows that the recombination effect is negligible in the presence of dust-neutral collisions. In line of this present situation our investigation revealed that the recombination is more important than dust-neutral collisions in laboratory plasma and fusion plasma, while the dust-neutral collision frequency is dominant in the interstellar plasmas. The effects of ion and dust densities and ion streaming on the recombination and collision driven mode in parameter regimes relevant for many experimental studies on dusty plasmas have also been calculated.

  5. Bimolecular recombination in organic photovoltaics.

    Science.gov (United States)

    Lakhwani, Girish; Rao, Akshay; Friend, Richard H

    2014-01-01

    The recombination of electrons and holes is a major loss mechanism in photovoltaic devices that controls their performance. We review scientific literature on bimolecular recombination (BR) in bulk heterojunction organic photovoltaic devices to bring forward existing ideas on the origin and nature of BR and highlight both experimental and theoretical work done to quantify its extent. For these systems, Langevin theory fails to explain BR, and recombination dynamics turns out to be dependent on mobility, temperature, electric field, charge carrier concentration, and trapped charges. Relationships among the photocurrent, open-circuit voltage, fill factor, and morphology are discussed. Finally, we highlight the recent emergence of a molecular-level picture of recombination, taking into account the spin and delocalization of charges. Together with the macroscopic picture of recombination, these new insights allow for a comprehensive understanding of BR and provide design principles for future materials and devices.

  6. In vitro biofumigation with Brassica juncea and Sinapis alba. Inhibition of germination and growth seedlings of weeds

    OpenAIRE

    Perniola, Omar Salvador; Chorzempa, Silvia Elena; Staltari, Sebastián; Molina, María del Carmen

    2016-01-01

    La biofumigación consiste en la incorporación en el suelo de residuos orgánicos, que generan durante su descomposición sustancias con actividad biocida. El objetivo de este trabajo fue determinar in vitro el efecto herbicida de la biofumigación con Brassica juncea L. Czerniak (mostaza parda) y Sinapis alba L. (mostaza blanca), en el estadio de fin de fructificación, sobre la germinación y el crecimiento de plántulas de tres especies arvenses: Anoda cristata (L.) Schltdl. (malva cimarrona), Pi...

  7. Measure Lines

    Science.gov (United States)

    Crissman, Sally

    2011-01-01

    One tool for enhancing students' work with data in the science classroom is the measure line. As a coteacher and curriculum developer for The Inquiry Project, the author has seen how measure lines--a number line in which the numbers refer to units of measure--help students not only represent data but also analyze it in ways that generate…

  8. Expression, purification, and functional characterization of recombinant PTD-SARA

    Institute of Scientific and Technical Information of China (English)

    Chen Huang; Rui Du; Peng Zhang; Hua Meng; Huiwei Jia; Yang Song; Man Li; Yingqi Zhang; Shiren Sun

    2011-01-01

    The Smad anchor for receptor activation (SARA) protein is a binding partner for Smad2/3 that plays an important role in the fibrotic promoting signaling pathway initiated by transforming growth factor-β1 (TGF-β1). The C-terminal 665-750 aa of SARA comprises the Smad-binding domain (SBD). By direct interaction through the SBD, SARA inhibits Smad2/3 phosphorylation and blocks the interaction between Smad2/3 and Smad4, thereby restrains the process of fibrosis.In this study, we constructed a SARA peptide aptamer based on the SBD sequence. The recombinant SARA aptamer,fused with a protein transduction domain (PTD-SARA), was cloned, purified from E. coli, and characterized for the first time. The full-length PTD-SARA coding sequence, created with E. coli favored codons, was cloned into a pQE-30 vector,and the recombinant plasmid was transformed into an M15 strain. After Isopropyl β-D-1-Thiogalactopyranoside (IPTG) induction and Ni2+ affinity purification, recombinant PTD-SARA was further identified by immunobiotting and protein N-terminal sequencing. Epifluorescence microscopy revealed that the recombinant PTD-SARA was transferred into the cytoplasm and nucleus more efficiently than SARA.Moreover, the recombinant PTD-SARA was found to up-regulate the level of E-cadherin and down-regulate the levels of α-SMA and phospho-Smad3 more efficiently than SARA (P< 0.05). Our work explored a method to obtain recombinant PTD-SARA protein. The recombinant PTDSARA fusion protein could enter HK2 cells (an immortalized proximal tubule epithelial cell line) more efficiently than the SARA protein and reverse the renal epithelial-to-mesenchymal transdifferentiation process that was induced by TGF-β1 more effectively than the SARA protein. Recombinant PDT-SARA is likely to be a potential candidate for clinical prevention and treatment of renal fibrosis.

  9. Homologous recombination efficiency enhanced by inhibition of MEK and GSK3β.

    Science.gov (United States)

    Lin, Zhaoyu; Zhang, Yanli; Gao, Tianyun; Wang, Liudi; Zhang, Qing; Zhou, Juan; Zhao, Jing

    2014-11-01

    Homologous recombination in embryonic stem cells (ESCs) is widely utilized in genome engineering, particularly in the generation of gene targeted mice. However, genome engineering is often plagued by the problem of low homologous recombination efficiency. In this study, we developed a novel method to increase the efficiency of homologous recombination in ESCs by changing its culture conditions. By comparing the efficiency of different ESCs in various culture conditions, we determined that chemicals that inhibit the MEK and GSK3β pathways (2i condition) enhance homologous recombination and eliminate differences in efficiencies among cell lines. Analysis of gene expression patterns in ESCs maintained in different culture conditions has identified several homologous recombination-related candidates, including the pluripotent markers Eras and Tbx3. The results of this study suggest that homologous recombination is associated with ESC pluripotency. © 2014 Wiley Periodicals, Inc.

  10. 土壤汞污染对油菜的氧化胁迫效应∗%Oxidative stress induced by mercury contaminated soil on Brassica juncea

    Institute of Scientific and Technical Information of China (English)

    陈礼洪; 赵康平; 蒋柱武; 刘学平

    2015-01-01

    In order to understand the phytotoxicity induced by mercury contaminated soil, the oxidative stress and mechanisms of Brassica juncea induced by mercury were investigated. It was shown that the low concentration mercury slightly promoted the growth of rapes, while rapes growth was significantly inhibited under the treatment of 1. 0 mg·kg-1 mercury. With increasing mercury concentration, malondialdehyde ( MDA) content, superoxide dismutase ( SOD) , peroxidase ( POD) and catalase ( CAT) activity increased at first and then decreased, indicating the oxidative stress. The accumulation of mercury increased with Hg concentration in soil, and the root is the main part of mercury accumulation. In the phytoremediation the root of Brassica juncea should be removed from the soil.%为了明确土壤汞污染的植物毒性效应,利用盆栽模拟试验研究了汞污染土壤对油菜的氧化损伤.结果表明,低浓度的汞污染轻微促进油菜的生长,汞浓度超过1.0 mg·kg-1就显著抑制植物生长,引起叶片面积变小、失绿变黄甚至枯萎等生理现象.随着汞浓度的增加,丙二醛( MDA )含量增加,当汞的浓度增加到8.0 mg·kg-1时,MDA的含量是对照组的4.17倍.随着汞浓度的增加,超氧化物歧化酶( SOD)活性升高,过氧化物酶(POD),过氧化氢酶(CAT)活性先升高后下降,其中SOD和POD在8.0 mg·kg-1汞处理下,活性达到最大,CAT在0.5 mg·kg-1汞处理下活性最大.土壤汞含量为超过1.0 mg·kg-1时,油菜可食部分汞浓度大于食品安全国家标准《食品中污染物限量》( GB 2762—2012)的规定.油菜吸收的汞主要累积在根部,因此在利用此类植物进行土壤修复时,需要清除植物的地下部,才能实现植物修复的目的.

  11. Thiourea, a ROS scavenger, regulates source-to-sink relationship to enhance crop yield and oil content in Brassica juncea (L..

    Directory of Open Access Journals (Sweden)

    Manish Pandey

    Full Text Available In the present agricultural scenario, the major thrust is to increase crop productivity so as to ensure sustainability. In an earlier study, foliar application of thiourea (TU; a non physiological thiol based ROS scavenger has been demonstrated to enhance the stress tolerance and yield of different crops under field condition. Towards this endeavor, present work deals with the effect of TU on photosynthetic efficiency and source-to-sink relationship of Indian mustard (Brassica juncea for understanding its mode of action. The application of TU increased the efficiency of both PSI and PSII photosystems and vegetative growth of plant. The comparative analysis of sucrose to starch ratio and expression level of sugar transporters confirmed the higher source and sink strength in response to TU treatment. The biochemical evidence in support of this was derived from higher activities of sucrose phosphate synthase and fructose-1,6-bis-phosphatase at source; and sucrose synthase and different classes of invertases at both source and sink. This indicated an overall increase in photoassimilate level at sink. An additional contribution through pod photosynthesis was confirmed through the analysis of phosphoenol pyruvate carboxylase enzyme activity and level of organic acids. The increased photoassimilate level was also co-ordinated with acetyl coA carboxylase mediated oil biosynthesis. All these changes were ultimately reflected in the form of 10 and 20% increase in total yield and oil content, respectively under TU treatment as compared to control. Additionally, no change was observed in oil composition of seeds derived from TU treated plants. The study thus signifies the co-ordinated regulation of key steps of photosynthesis and source-to-sink relationship through the external application of TU resulting in increased crop yield and oil content.

  12. Characterization of interploid hybrids from crosses between Brassica juncea and B. oleracea and the production of yellow-seeded B. napus.

    Science.gov (United States)

    Wen, Jing; Zhu, Lixia; Qi, Liping; Ke, Hongmei; Yi, Bin; Shen, Jinxiong; Tu, Jinxing; Ma, Chaozhi; Fu, Tingdong

    2012-06-01

    Yellow-seeded Brassica napus was for the first time developed from interspecific crosses using yellow-seeded B. juncea (AABB), yellow-seeded B. oleracea (CC), and black-seeded artificial B. napus (AACC). Three different mating approaches were undertaken to eliminate B-genome chromosomes after trigenomic hexaploids (AABBCC) were generated. Hybrids (AABCC, ABCC) from crosses AABBCC × AACC, AABBCC × CC and ABCC × AACC were advanced by continuous selfing in approach 1, 2 and 3, respectively. To provide more insight into Brassica genome evolution and the cytological basis for B. napus resynthesis in each approach, B-genome chromosome pairing and segregation were intensively analyzed in AABCC and ABCC plants using genomic in situ hybridization methods. The frequencies at which B-genome chromosomes underwent autosyndesis and allosyndesis were generally higher in ABCC than in AABCC plants. The difference was statistically significant for allosyndesis but not autosyndesis. Abnormal distributions of B-genome chromosomes were encountered at anaphase I, including chromosome lagging and precocious sister centromere separation of univalents. These abnormalities were observed at a significantly higher frequency in AABCC than in ABCC plants, which resulted in more rapid B-genome chromosome elimination in the AABCC derivatives. Yellow or yellow-brown seeds were obtained in all approaches, although true-breeding yellow-seeded B. napus was developed only in approaches 2 and 3. The efficiency of the B. napus construction approaches was in the order 1 > 3 > 2 whereas this order was 3 > 2 > 1 with respect to the construction of yellow-seeded B. napus. The results are discussed in relation to Brassica genome evolution and the development and utilization of the yellow-seeded B. napus obtained here.

  13. Analysis of interchromosomal mitotic recombination.

    Science.gov (United States)

    McGill, C B; Shafer, B K; Higgins, D R; Strathern, J N

    1990-07-01

    A novel synthetic locus is described that provides a simple assay system for characterizing mitotic recombinants. The locus consists of the TRP1 and HIS3 genes inserted into chromosome III of S. cerevisiae between the CRY1 and MAT loci. Defined trp1 and his3 alleles have been generated that allow the selection of interchromosomal recombinants in this interval. Trp+ or His+ recombinants can be divided into several classes based on coupling of the other alleles in the interval. The tight linkage of the CRY1 and MAT loci, combined with the drug resistance and cell type phenotypes that they respectively control, facilitates the classification of the recombinants without resorting to tetrad dissection. We present the distribution of spontaneous recombinants among the classes defined by this analysis. The data suggest that the recombination intermediate can have regions of symmetric strand exchange and that co-conversion tracts can extend over 1-3 kb. Continuous conversion tracts are favored over discontinuous tracts. The distribution among the classes defined by this analysis is altered in recombinants induced by UV irradiation.

  14. Testing for recombinant erythropoietin.

    Science.gov (United States)

    Delanghe, Joris R; Bollen, Mathieu; Beullens, Monique

    2008-03-01

    Erythropoietin (Epo) is a glycoprotein hormone that promotes the production of red blood cells. Recombinant human Epo (rhEpo) is illicitly used to improve performance in endurance sports. Doping in sports is discouraged by the screening of athletes for rhEpo. Both direct tests (indicating the presence of exogeneous Epo isoforms) and indirect tests (indicating hematological changes induced by exogenous Epo administration) can be used for Epo detection. At present, the test adopted by the World Anti Doping Agency is based on a combination of isoelectric focusing and double immunoblotting, and distinguishes between endogenous and rhEpo. However, the adopted monoclonal anti-Epo antibodies are not monospecific. Therefore, the test can occasionally lead to the false-positive detection of rhEpo (epoetin-beta) in post-exercise, protein-rich urine, or in case of contamination of the sample with microorganisms. An improved preanalytical care may counteract a lot of these problems. Adaptation of the criteria may be helpful to further refine direct Epo testing. Indirect tests have the disadvantage that they require blood instead of urine samples, but they can be applied to detect a broader range of performance improving techniques which are illicitly used in sports.

  15. Monitoring of recombinant protein production using bioluminescence in a semiautomated fermentation process.

    Science.gov (United States)

    Trezzani, I; Nadri, M; Dorel, C; Lejeune, P; Bellalou, J; Lieto, J; Hammouri, H; Longin, R; Dhurjati, P

    2003-01-01

    On-line optimization of fermentation processes can be greatly aided by the availability of information on the physiological state of the cell. The goal of our "BioLux" research project was to design a recombinant cell capable of intracellular monitoring of product synthesis and to use it as part of an automated fermentation system. A recombinant plasmid was constructed containing an inducible promoter that controls the gene coding for a model protein and the genes necessary for bioluminescence. The cells were cultured in microfermenters equipped with an on-line turbidity sensor and a specially designed on-line light sensor capable of continuous measurement of bioluminescence. Initial studies were done under simple culture conditions, and a linear correlation between luminescence and protein production was obtained. Such specially designed recombinant bioluminescent cells can potentially be applied for model-based inference of intracellular product formation, as well as for optimization and control of recombinant fermentation processes.

  16. Controlled release from recombinant polymers.

    Science.gov (United States)

    Price, Robert; Poursaid, Azadeh; Ghandehari, Hamidreza

    2014-09-28

    Recombinant polymers provide a high degree of molecular definition for correlating structure with function in controlled release. The wide array of amino acids available as building blocks for these materials lend many advantages including biorecognition, biodegradability, potential biocompatibility, and control over mechanical properties among other attributes. Genetic engineering and DNA manipulation techniques enable the optimization of structure for precise control over spatial and temporal release. Unlike the majority of chemical synthetic strategies used, recombinant DNA technology has allowed for the production of monodisperse polymers with specifically defined sequences. Several classes of recombinant polymers have been used for controlled drug delivery. These include, but are not limited to, elastin-like, silk-like, and silk-elastinlike proteins, as well as emerging cationic polymers for gene delivery. In this article, progress and prospects of recombinant polymers used in controlled release will be reviewed.

  17. Cell encoding recombinant human erythropoietin

    Energy Technology Data Exchange (ETDEWEB)

    Beck, A.K.; Withy, R.M.; Zabrecky, J.R.; Masiello, N.C.

    1990-09-04

    This patent describes a C127 cell transformed with a recombinant DNA vector. It comprises: a DNA sequence encoding human erythropoietin, the transformed cell being capable of producing N-linked and O-linked glycosylated human erythropoietin.

  18. Influenza Vaccine, Inactivated or Recombinant

    Science.gov (United States)

    ... die from flu, and many more are hospitalized.Flu vaccine can:keep you from getting flu, make flu ... inactivated or recombinant influenza vaccine?A dose of flu vaccine is recommended every flu season. Children 6 months ...

  19. Three Decades of Recombinant DNA.

    Science.gov (United States)

    Palmer, Jackie

    1985-01-01

    Discusses highlights in the development of genetic engineering, examining techniques with recombinant DNA, legal and ethical issues, GenBank (a national database of nucleic acid sequences), and other topics. (JN)

  20. Perovskite photovoltaics: Slow recombination unveiled

    Science.gov (United States)

    Moser, Jacques-E.

    2017-01-01

    One of the most salient features of hybrid lead halide perovskites is the extended lifetime of their photogenerated charge carriers. This property has now been shown experimentally to originate from a slow, thermally activated recombination process.

  1. Inhomogeneous recombinations during cosmic reionization

    OpenAIRE

    Sobacchi, Emanuele; Mesinger, Andrei

    2014-01-01

    By depleting the ionizing photon budget available to expand cosmic HII regions, recombining systems (or Lyman limit systems) can have a large impact during (and following) cosmic reionization. Unfortunately, directly resolving such structures in large-scale reionization simulations is computationally impractical. Instead, here we implement a sub-grid prescription for tracking inhomogeneous recombinations in the intergalactic medium. Building on previous work parameterizing photo-heating feedb...

  2. Plasmid recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.

    1982-01-01

    DNA recombination in exponential phase and competent Haemophilus influenzae was measured by an electron microscopic assay that relies on the conversion of plasmid RSF0885 monomers into multimeric forms. Dimer circles were present at a frequency of 2% in plasmid preparations from competent Rd (wild-type) cells; multimers were present at a frequency of 0.2% in preparations from exponential phase cells. Thus, plasmid recombination was stimulated in competent cells. Multimer formation occurred efficiently in cells of the transformation defective mutant rec2, implying that the rec2 gene product is not required for plasmid recombination. However, the absence of multimer plasmids in preparations from competent cells of the transformation defective mutant rec1 suggests that the rec1 gene product is required. Digestion of purified plasmids with restriction endonuclease PvuII, which makes a single cut in the monomer, revealed the presence of recombination intermediates composed of two linear plasmids joined to form two pairs of arms resembling the Greek letter chi. Length measurements of these arms taken from a population of recombination intermediates gave evidence that the plasmids were joined at sites of homology. The distributions of individual DNA strands, at the intersections of the four arms, could be resolved in some recombination intermediates and were of two types. The first type of junction appeared as a single-stranded arm appended to each corner. The second type of junction consisted of a single strand of DNA linking the two linear plasmids at a site of homology. The single-stranded linker was frequently situated at the edge of a short gap on one of the plasmids in the pair. The fine structures of the recombinational joints have been interpreted in terms of previously proposed models of recombination.

  3. Heterogeneity in recombinant protein production

    DEFF Research Database (Denmark)

    Schalén, Martin; Johanson, Ted; Lundin, Luisa;

    2012-01-01

    contribute to make a population in a fermenter heterogeneous, resulting in cell-to-cell variation in physiological parameters of the microbial culture. Our study aims at investigating how population heterogeneity and recombinant protein production is affected by environmental gradients in bioreactors...... are simulated in small bioreactors and the population heterogeneity can be visualised by analysing single cells with flow cytometry. This can give new insights to cell physiology and recombinant protein production at the industrial scale....

  4. Genetic Analysis of Trichome Density on the Main Stem and Leaves in a Recombinant Inbred Lines Population Derived from Wild Pepper (Capsicum annuum L.)%野生辣椒重组自交系群体主茎和叶片表面茸毛密度的遗传分析

    Institute of Scientific and Technical Information of China (English)

    张晓芬; 陈晓慧; 陈斌; 韩华丽; 耿三省

    2013-01-01

    Plant trichomes are attracting more and more attention for their effect on stress resistance. A F9 recombinant inbred lines population derived from the cross of wild hot pepper(Capsicum annuum L.) PM702 bearing trichomes with glabrous sweet pepper(C. annuum L.) FS871, was used for genetic analysis of trichome density on the main stem and leaves, using the mixed major gene plus polygene inheritance model. The results showed that trichome density on the main stem of pepper was controlled by two major genes and some minor polygenes(E-2-3 model). For the two genes, the additive effect was primary. The heritability of major genes for the trichome density on the main stem was 53.00%, while the heritability of polygenes was 25.30%, and the recombination rate was 0.6226. The trichome density on the leaves was controlled by two complementary major genes with additive effect and interaction plus some polygenes(E-1-7 model). The heritability of major genes of trichome density on the leaves was 50.65%, while the heritability of polygenes was 8.86%. This study provides an academic foundation for breeding highly resistant pepper varieties.%植株茎叶茸毛在抗逆性所起的作用正在引起人们越来越多的关注.本研究以多毛辣椒(Capsicum annuum L.)PM702和无毛甜椒(C.annuum L.)FS871为双亲构建的F9代重组自交系(RILs)为实验材料,利用主基因+多基因混合遗传模型,联合双亲对主茎和叶片表面茸毛密度进行了遗传分析.结果显示,辣椒主茎表面茸毛密度分布符合E-2-3模型,即受2对连锁的主基因控制,并有多基因的修饰.2对主基因以加性效应为主.主基因遗传率为53.00%,多基因遗传率为25.30%.两基因座间的重组率r=0.6226.叶片正面茸毛密度分布符合E-1-7遗传模型,即受2对具有互补作用的主基因控制,并有多基因同时起作用.主基因以加性效应为主,并有互作.主基因遗传率50.65%,多基因遗传率8.86%.结果为抗逆甜辣椒

  5. Recombinant protein expression in Nicotiana.

    Science.gov (United States)

    Matoba, Nobuyuki; Davis, Keith R; Palmer, Kenneth E

    2011-01-01

    Recombinant protein pharmaceuticals are now widely used in treatment of chronic diseases, and several recombinant protein subunit vaccines are approved for human and veterinary use. With growing demand for complex protein pharmaceuticals, such as monoclonal antibodies, manufacturing capacity is becoming limited. There is increasing need for safe, scalable, and economical alternatives to mammalian cell culture-based manufacturing systems, which require substantial capital investment for new manufacturing facilities. Since a seminal paper reporting immunoglobulin expression in transgenic plants was published in 1989, there have been many technological advances in plant expression systems to the present time where production of proteins in leaf tissues of nonfood crops such as Nicotiana species is considered a viable alternative. In particular, transient expression systems derived from recombinant plant viral vectors offer opportunities for rapid expression screening, construct optimization, and expression scale-up. Extraction of recombinant proteins from Nicotiana leaf tissues can be achieved by collection of secreted protein fractions, or from a total protein extract after grinding the leaves with buffer. After separation from solids, the major purification challenge is contamination with elements of the photosynthetic complex, which can be solved by application of a variety of facile and proven strategies. In conclusion, the technologies required for safe, efficient, scalable manufacture of recombinant proteins in Nicotiana leaf tissues have matured to the point where several products have already been tested in phase I clinical trials and will soon be followed by a rich pipeline of recombinant vaccines, microbicides, and therapeutic proteins.

  6. The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection.

    Directory of Open Access Journals (Sweden)

    Qingwen Jin

    Full Text Available Insertion of T4 lysozyme (T4L into the GPCR successfully enhanced GPCR protein stability and solubilization. However, the biological functions of the recombinant GPCR protein have not been analyzed.We engineered the CCR5-T4L mutant and expressed and purified the soluble recombinant protein using an E.coli expression system. The antiviral effects of this recombinant protein in THP-1 cell lines, primary human macrophages, and PBMCs from different donors were investigated. We also explored the possible mechanisms underlying the observed antiviral effects.Our data showed the biphasic inhibitory and promotion effects of different concentrations of soluble recombinant CCR5-T4L protein on R5 tropic human immunodeficiency virus-1 (HIV-1 infection in THP-1 cell lines, human macrophages, and PBMCs from clinical isolates. We demonstrated that soluble recombinant CCR5-T4L acts as a HIV-1 co-receptor, interacts with wild type CCR5, down-regulates the surface CCR5 expression in human macrophages, and interacts with CCL5 to inhibit macrophage migration. Using binding assays, we further determined that recombinant CCR5-T4L and [125I]-CCL5 compete for the same binding site on wild type CCR5.Our results suggest that recombinant CCR5-T4L protein marginally promotes HIV-1 infection at low concentrations and markedly inhibits infection at higher concentrations. This recombinant protein may be helpful in the future development of anti-HIV-1 therapeutic agents.

  7. Something new about recombination

    CERN Document Server

    Rijo, Juan Betancort

    2013-01-01

    In the seminal works of Zeldovich (1968) and Peebles (1968), a procedure was outlined to obtain the equation of evolution of the hydrogen fraction without an explicit use of the radiative transfer equation. This procedure is based, explicitly or implicitly, on Sobolev (1960) escape probability and has extensively been used since then in developing refined approximations. Here we present a somewhat different derivation of that procedure that is both simple and formally exact, and use it to assess the implications of a number of effects. The most remarkable one is the fact that a photon that has escaped from the line 2p -> 1s can be absorbed, conjointly with another photon with the appropriate frequency, by the two photon line 1s -> 2s. This effect, which apparently has not been treated before, implies a maximum increment in the electronic fraction of roughly a 2 % at z around 1100 which is large enough to have non-trivial consequences on the inference of cosmological parameters from CMB maps.

  8. Recombinational DNA repair and human disease

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, Larry H.; Schild, David

    2002-11-30

    We review the genes and proteins related to the homologous recombinational repair (HRR) pathway that are implicated in cancer through either genetic disorders that predispose to cancer through chromosome instability or the occurrence of somatic mutations that contribute to carcinogenesis. Ataxia telangiectasia (AT), Nijmegen breakage syndrome (NBS), and an ataxia-like disorder (ATLD), are chromosome instability disorders that are defective in the ataxia telangiectasia mutated (ATM), NBS, and Mre11 genes, respectively. These genes are critical in maintaining cellular resistance to ionizing radiation (IR), which kills largely by the production of double-strand breaks (DSBs). Bloom syndrome involves a defect in the BLM helicase, which seems to play a role in restarting DNA replication forks that are blocked at lesions, thereby promoting chromosome stability. The Werner syndrome gene (WRN) helicase, another member of the RecQ family like BLM, has very recently been found to help mediate homologous recombination. Fanconi anemia (FA) is a genetically complex chromosomal instability disorder involving seven or more genes, one of which is BRCA2. FA may be at least partially caused by the aberrant production of reactive oxidative species. The breast cancer-associated BRCA1 and BRCA2 proteins are strongly implicated in HRR; BRCA2 associates with Rad51 and appears to regulate its activity. We discuss in detail the phenotypes of the various mutant cell lines and the signaling pathways mediated by the ATM kinase. ATM's phosphorylation targets can be grouped into oxidative stress-mediated transcriptional changes, cell cycle checkpoints, and recombinational repair. We present the DNA damage response pathways by using the DSB as the prototype lesion, whose incorrect repair can initiate and augment karyotypic abnormalities.

  9. Laminin-121--recombinant expression and interactions with integrins.

    Science.gov (United States)

    Sasaki, Takako; Takagi, Junichi; Giudici, Camilla; Yamada, Yoshihiko; Arikawa-Hirasawa, Eri; Deutzmann, Rainer; Timpl, Rupert; Sonnenberg, Arnoud; Bächinger, Hans Peter; Tonge, David

    2010-07-01

    Laminin-121, previously referred as to laminin-3, was expressed recombinantly in human embryonic kidney (HEK) 293 cells by triple transfection of full-length cDNAs encoding mouse laminin α1, β2 and γ1 chains. The recombinant laminin-121 was purified using Heparin-Sepharose followed by molecular sieve chromatography and shown to be correctly folded by electron microscopy and circular dichroism (CD). The CD spectra of recombinant laminin-121 were very similar to those of laminin-111 isolated from Engelbreth-Holm-Swarm tumor (EHS-laminin) but its T(m) value was smaller than EHS-laminin and recombinant lamnin-111 suggesting that the replacement of the β chain reduced the stability of the coiled-coil structure of laminin-121. Its binding to integrins was compared with EHS-laminin, laminin-3A32 purified from murine epidermal cell line and recombinantly expressed laminins-111, -211 and -221. Laminin-121 showed the highest affinity to α6β1 and α7β1 integrins and furthermore, laminin-121 most effectively supported neurite outgrowth. Together, this suggests that the β2 laminins have higher affinity for integrins than the β1 laminins.

  10. RECOMBINANT HORSERADISH PEROXIDASE FOR ANALYTICAL APPLICATIONS

    Directory of Open Access Journals (Sweden)

    А.M. Egorov

    2012-08-01

    Full Text Available The article deals with prospects of using recombinant horseradish peroxidase in analytical biochemistry and biotechnology. Problems of recombinant horseradish peroxidase cloning in different expression systems, possible approaches to their solution, advantages of recombinant recombinant horseradish peroxidase and recombinant horseradish peroxidase-fusion proteins for immunoassays are considered. Possibility for development of mediatorless bienzyme biosensor for peroxide and metabolites, yielding hydrogen peroxide during their transformations, based on co-adsorption of recombinant horseradish peroxidase and the appropriate oxidase was demonstrated. The possibility to produce a fully active recombinant conjugate of recombinant horseradish peroxidase with human heart-type fatty acid binding protein, which may be used in competitive immunoassay for clinical diagnosis of acute myocardial infarction, and recombinant conjugates (N- and C-terminus of recombinant horseradish peroxidase with Fab-fragments of the antibody against atrazine, which may be applied for atrazine pesticides detection, are demonstra ted for the first time.

  11. Recent advances in recombinant protein production

    Science.gov (United States)

    Kunert, Renate; Casanova, Emilio

    2013-01-01

    Designing appropriate expression vectors is one of the critical steps in the generation of stable cell lines for recombinant protein production. Conventional expression vectors are severely affected by the chromatin environment surrounding their integration site into the host genome, resulting in low expression levels and transgene silencing. In the past, a new generation of expression vectors and different strategies was developed to overcome the chromatin effects. Bacterial artificial chromosomes (BACs) are cloning vectors capable of accommodating up to 350 Kb. Thus, BACs can carry a whole eukaryotic locus with all the elements controlling the expression of a gene; therefore, BACs harbor their own chromatin environment. Expression vectors based on BACs containing open/permissive chromatin loci are not affected by the chromatin surrounding their integration site in the host cell genome. Consequently, BAC-based expression vectors containing the appropriate loci confer predictable and high levels of expression over time. These properties make BAC-based expression vectors a very attractive tool applied to the recombinant protein production field. PMID:23680894

  12. Recombinant shark natural antibodies to thyroglobulin.

    Science.gov (United States)

    Schluter, Samuel F; Jensen, Ingvill; Ramsland, Paul A; Marchalonis, John J

    2005-01-01

    As cartilaginous fish are the vertebrates most distal from man to produce antibodies, fundamental information regarding conservation and variation of the antigen binding site should be gained by comparing the properties of antibodies directed against the same antigen from the two species. Since monoclonal cell lines cannot be generated using shark B cells, we isolated antigen binding recombinant single chain Fv antibodies (scFv) comprising of the complete variable regions from shark light and heavy chains. Thyroglobulin was used as the selecting antigen as both sharks and humans express natural antibodies to mammalian thyroglobulin in the absence of purposeful immunization. We report that recombinant sandbar shark (Carcharhinus plumbeus) scFvs that bind bovine thyroglobulin consist of heavy chain variable regions (VH) homologous to those of the human VHIII subset and light chain variable regions (VL) homologous to those of the human Vlambda6 subgroup. The homology within the frameworks is sufficient to enable the building of three-dimensional models of the shark VH/VL structure using established human structures as templates. In natural antibodies of both species, the major variability lies in the third complementarity determining region (CDR3) of both VH and VL.

  13. Recombination drives vertebrate genome contraction.

    Directory of Open Access Journals (Sweden)

    Kiwoong Nam

    Full Text Available Selective and/or neutral processes may govern variation in DNA content and, ultimately, genome size. The observation in several organisms of a negative correlation between recombination rate and intron size could be compatible with a neutral model in which recombination is mutagenic for length changes. We used whole-genome data on small insertions and deletions within transposable elements from chicken and zebra finch to demonstrate clear links between recombination rate and a number of attributes of reduced DNA content. Recombination rate was negatively correlated with the length of introns, transposable elements, and intergenic spacer and with the rate of short insertions. Importantly, it was positively correlated with gene density, the rate of short deletions, the deletion bias, and the net change in sequence length. All these observations point at a pattern of more condensed genome structure in regions of high recombination. Based on the observed rates of small insertions and deletions and assuming that these rates are representative for the whole genome, we estimate that the genome of the most recent common ancestor of birds and lizards has lost nearly 20% of its DNA content up until the present. Expansion of transposable elements can counteract the effect of deletions in an equilibrium mutation model; however, since the activity of transposable elements has been low in the avian lineage, the deletion bias is likely to have had a significant effect on genome size evolution in dinosaurs and birds, contributing to the maintenance of a small genome. We also demonstrate that most of the observed correlations between recombination rate and genome contraction parameters are seen in the human genome, including for segregating indel polymorphisms. Our data are compatible with a neutral model in which recombination drives vertebrate genome size evolution and gives no direct support for a role of natural selection in this process.

  14. Conservation of recombination hotspots in yeast

    OpenAIRE

    Tsai, Isheng J.; Burt, Austin; Koufopanou, Vassiliki

    2010-01-01

    Meiotic recombination does not occur randomly along a chromosome, but instead tends to be concentrated in small regions, known as “recombination hotspots.” Recombination hotspots are thought to be short-lived in evolutionary time due to their self-destructive nature, as gene conversion favors recombination-suppressing alleles over recombination-promoting alleles during double-strand repair. Consistent with this expectation, hotspots in humans are highly dynamic, with little correspondence in ...

  15. Recombination at the DNA level. Abstracts

    Energy Technology Data Exchange (ETDEWEB)

    1984-01-01

    Abstracts of papers in the following areas are presented: (1) chromosome mechanics; (2) yeast systems; (3) mammalian homologous recombination; (4) transposons; (5) Mu; (6) plant transposons/T4 recombination; (7) topoisomerase, resolvase, and gyrase; (8) Escherichia coli general recombination; (9) recA; (10) repair; (11) eucaryotic enzymes; (12) integration and excision of bacteriophage; (13) site-specific recombination; and (14) recombination in vitro. (ACR)

  16. Transgenic Expression of the Recombinant Phytase in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    LIU Qiao-quan; LI Qian-feng; JIANG Li; ZHANG Da-jiang; WANG Hong-mei; GU Ming-hong; YAO Quan-hong

    2006-01-01

    In most of the cereal crop, phytic acid is the main storage form of phosphorus, which can decrease the bioavailability of phosphate. Transgenic expression of phytase is regarded as an efficient way to release phosphate from phytate in transgenic plants.In this study, a plant expression vector, containing the recombinant phytase gene driven by the maize ubiquitin (Ubi) promoter was constructed and introduced into an elite rice variety via Agrobacterium-mediated transformation. During the experiment, a total of 15 independent transgenic rice lines were regenerated. The results of PCR and Southern blot indicated that the target gene was integrated into the genome of transgenic rice plants. Moreover, the RT-PCR analysis of total RNAs extracted from the immature seeds of several transgenic lines showed that the recombinant phytase gene could be normally expressed. The inorganic phosphorus content, both in the mature seeds and the leaf was significantly higher in the transgenic plants than in the untransformed wild type.

  17. Serendipitous identification of natural intergenotypic recombinants of hepatitis C in Ireland.

    LENUS (Irish Health Repository)

    Moreau, Isabelle

    2006-01-01

    BACKGROUND: Recombination between hepatitis C single stranded RNA viruses is a rare event. Natural viable intragenotypic and intergenotypic recombinants between 1b-1a, 1a-1c and 2k-1b, 2i-6p, respectively, have been reported. Diagnostically recombinants represent an intriguing challenge. Hepatitis C genotype is defined by interrogation of the sequence composition of the 5\\' untranslated region [5\\'UTR]. Occasionally, ambiguous specimens require further investigation of the genome, usually by interrogation of the NS5B region. The original purpose of this study was to confirm the existence of a suspected mixed genotype infection of genotypes 2 and 4 by clonal analysis at the NS5B region of the genome in two specimens from two separate individuals. This initial identification of genotype was based on analysis of the 5\\'UTR of the genome by reverse line probe hybridisation [RLPH]. RESULTS: The original diagnosis of a mixed genotype infection was not confirmed by clonal analysis of the NS5B region of the genome. The phylogenetic analysis indicated that both specimens were natural intergenotypic recombinant forms of HCV. The recombination was between genotypes 2k and 1b for both specimens. The recombination break point was identified as occurring within the NS2 region of the genome. CONCLUSION: The viral recombinants identified here resemble the recombinant form originally identified in Russia. The RLPH pattern observed in this study may be a signature indicative of this particular type of intergenotype recombinant of hepatitis C meriting clonal analysis of NS2.

  18. Recombinant allergens for pollen immunotherapy.

    Science.gov (United States)

    Wallner, Michael; Pichler, Ulrike; Ferreira, Fatima

    2013-12-01

    Specific immunotherapy (IT) represents the only potentially curative therapeutic intervention of allergic diseases capable of suppressing allergy-associated symptoms not only during treatment, but also after its cessation. Presently, IT is performed with allergen extracts, which represent a heterogeneous mixture of allergenic, as well as nonallergenic, compounds of a given allergen source. To overcome many of the problems associated with extract-based IT, strategies based on the use of recombinant allergens or derivatives thereof have been developed. This review focuses on recombinant technologies to produce allergy therapeuticals, especially for allergies caused by tree, grass and weed pollen, as they are among the most prevalent allergic disorders affecting the population of industrialized societies. The reduction of IgE-binding of recombinant allergen derivatives appears to be mandatory to increase the safety profile of vaccine candidates. Moreover, increased immunogenicity is expected to reduce the dosage regimes of the presently cumbersome treatment. In this regard, it has been convincingly demonstrated in animal models that hypoallergenic molecules can be engineered to harbor inherent antiallergenic immunologic properties. Thus, strategies to modulate the allergenic and immunogenic properties of recombinant allergens will be discussed in detail. In recent years, several successful clinical studies using recombinant wild-type or hypoallergens as active ingredients have been published and, currently, novel treatment forms with higher safety and efficacy profiles are under investigation in clinical trials. These recent developments are summarized and discussed.

  19. Transfection of recombinant retrovirus vector pLXSN-glial cell line-derived neurotrophic factor into umbilical cord-derived mesenchymal stem cells%pLXSN-胶质细胞源性神经营养因子重组载体转染脐带间充质干细胞的研究

    Institute of Scientific and Technical Information of China (English)

    吴学建; 韩克; 朱旭

    2012-01-01

    目的 构建大鼠胶质细胞源性神经营养因子(GDNF)基因修饰的脐带间充质干细胞(UCMSCs).方法 鉴定重组体中目的基因.脂质体包裹法将pLXSN-GDNF(携带大鼠胶质细胞源性生长因子的重组逆转录病毒载体)包装到PA317细胞中.NIH3T3细胞测定逆转录病毒滴度.病毒转染增殖旺盛的UCMSCs.免疫组织化学染色法和逆转录-聚合酶链反应(RT-PCR)法检测GDNF基因的表达.结果 目的基因正确.脂质体包裹法成功将pLXSN-GDNF载体转染入包装细胞PA317中.NIH3T3细胞测定最高病毒滴度为1×104 CFU/mt.免疫组织化学染色结果示:GDNF-UCMSCs抗GDNF蛋白染色阳性.RT-PCR结果示:转染后GDNF-UCMSCs表达GDNF mRNA的水平明显高于未转染的UCMSCs,差异有统计学意义(P<0.01).结论 成功构建GDNF基因修饰的UCMSCs.%Objective To construct human umbilical cord-derived mesenchymal stem cells which were modified by glial cell line-derived neurotrophic factor gene.Methods The target gene in was identified.PA317 cells were transfected with recombinant retroviral vector pLXSN-glial cell line-derived neurotrophic factor (GDNF) using liposomes.The retrovirus titers were determined.Then the umbilical cord-derived mesenchymal stem cells (UCMSCs) were infected by pLXSN-GDNF.Finally,the immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of GDNF.Results The target gene was correct.The pLXSN-GDNF vector was successfully transfected into the PA317 cells using liposomes.The highest virus titre of the clone was 1 x 104 CFU/ml.Cells in both groups were immunohistochemically positive for GDNF expression.Staining for GDNF was more prominent in the UCMSCs infected with pLXSN-GDNF.RT-PCR revealed the UCMSCs modified by GDNF gene expressed GDNF mRNA obviously higher than the UCMSCs which were not decorated by GDNF gene (P <0.01).Conclusion The UCMSCs modified by GDNF were successfully constructed.

  20. High Line

    DEFF Research Database (Denmark)

    Kiib, Hans

    2015-01-01

    At just over 10 meters above street level, the High Line extends three kilometers through three districts of Southwestern Manhattan in New York. It consists of simple steel construction, and previously served as an elevated rail line connection between Penn Station on 34th Street and the many....... The High Line project has been carried out as part of an open conversion strategy. The result is a remarkable urban architectural project, which works as a catalyst for the urban development of Western Manhattan. The greater project includes the restoration and reuse of many old industrial buildings...... in close proximity to the park bridge and new projects being added to fit the context. The outcome is a conglomeration of non-contemporary urban activities along the High Line, where mechanical workshops, small wholesale stores. etc. mix with new exclusive residential buildings, eminent cafés...

  1. High Line

    DEFF Research Database (Denmark)

    Kiib, Hans

    2015-01-01

    in close proximity to the park bridge and new projects being added to fit the context. The outcome is a conglomeration of non-contemporary urban activities along the High Line, where mechanical workshops, small wholesale stores. etc. mix with new exclusive residential buildings, eminent cafés......At just over 10 meters above street level, the High Line extends three kilometers through three districts of Southwestern Manhattan in New York. It consists of simple steel construction, and previously served as an elevated rail line connection between Penn Station on 34th Street and the many...... factories and warehouses on Gansevoort Street. Today the High Line is a beautiful park covered with new tiles, viewing platforms and smaller recreational areas. The park bridge has simple, uniform, urban fittings and features a variety of flowering plants, grasses, shrubs and trees from around the world...

  2. Helium recombination spectra as temperature diagnostics for planetary nebulae

    CERN Document Server

    Zhang, Y; Liu, Y; Rubin, R H

    2005-01-01

    Electron temperatures derived from the \\ion{He}{1} recombination line ratios, designated $T_{\\rm e}$(\\ion{He}{1}), are presented for 48 planetary nebulae (PNe). We study the effect that temperature fluctuations inside nebulae have on the $T_{\\rm e}$(\\ion{He}{1}) value. We show that a comparison between $T_{\\rm e}$(\\ion{He}{1}) and the electron temperature derived from the Balmer jump of the \\ion{H}{1} recombination spectrum, designated $T_{\\rm e}$(\\ion{H}{1}), provides an opportunity to discriminate between the paradigms of a chemically homogeneous plasma with temperature and density variations, and a two-abundance nebular model with hydrogen-deficient material embedded in diffuse gas of a ``normal'' chemical composition (i.e. $\\sim$ solar), as the possible causes of the dichotomy between the abundances that are deduced from collisionally excited lines to those deduced from recombination lines. We find that $T_{\\rm e}$(\\ion{He}{1}) values are significantly lower than $T_{\\rm e}$(\\ion{H}{1}) values, with an av...

  3. Evidence for natural recombination between mink enteritis virus and canine parvovirus

    Directory of Open Access Journals (Sweden)

    Wang Jianke

    2012-10-01

    Full Text Available Abstract A virus was isolated from mink showing clinical and pathological signs of enteritis in China. This virus, designated MEV/LN-10, was identified as mink enteritis virus (MEV based on its cytopathic effect in the feline F81 cell line, the hemagglutination (HA and hemagglutination inhibition (HI assay, electron microscopy (EM and animal infection experiments. The complete viral genome was cloned and sequenced. Phylogenetic and recombination analyses on the complete MEV/LN-10 genome showed evidence of recombination between MEV and canine parvovirus (CPV. The genome was composed of the NS1 gene originating from CPV while the VP1 gene was of MEV origin. This is the first demonstration of recombination between a CPV and MEV in nature. Our findings not only provide valuable evidence indicating that recombination is an important genetic mechanism contributing to the variation and evolution of MEV, but also that heterogeneous recombination can occur in the feline parvovirus subspecies.

  4. [Construction and characterization of a novel recombinant retroviral vector expressing mouse T-bet].

    Science.gov (United States)

    Zhang, Xuejie; Zhang, Jianhua; Zhang, Wei; Guo, Jie; Zhou, Xuyu

    2014-10-01

    In order to study T-bet function in mouse cells, a novel retroviral vector expressing mouse T-bet and reporter gene Thy1.1 was constructed. Retrovirus particles were then produced by transfection of the recombinant retroviral plasmid into a packaging cell line Platinum-E. The recombinant retrovirus played considerable infection ability. T-bet expression was then identified by FACS after infection of CD4+ primary T cells from T-bet knockout mouse with recombinant retrovirus. To determine if exogenous expressing T-bet has normal function, we checked the expression level of T-bet target gene, Ifng. IFN-y expression was upregulated in the T-bet knockout T cells infected with recombinant retrovirus. In conclusion, we successfully constructed an effective mouse T-bet recombinant retroviral vector.

  5. Creation of a completely helper cell-dependent recombinant morbillivirus

    OpenAIRE

    Baron, Jana; Baron, Michael

    2013-01-01

    We have created a completely helper cell-dependent morbillivirus by modifying the genome to remove the coding sequence of the phosphoprotein (P) and recovering the recombinant virus in a cell line constitutively expressing the P protein. The P protein-deleted virus (P−) grew very inefficiently unless both of the viral accessory proteins (V and C) were also expressed. Growth of the virus was restricted to the P-expressing cell line. The P− virus grew more slowly than the parental virus and exp...

  6. The effect of a single recombination event

    DEFF Research Database (Denmark)

    Schierup, Mikkel Heide; Jensen, Thomas Mailund; Wiuf, Carsten

    We investigate the variance in how visible a single recombination event is in a SNP data set as a function of the type of recombination event and its age. Data is simulated under the coalescent with recombination and inference is by the popular composite likelihood methods. The major determinant...... of the effect of a recombination event is the genealogical type of the event and whether SNP variation is present that can reveal the genealogical consequences of the recombination event. Recombination events that only change some branch lengths in the genealogy have a very small, but detectable, effect....... The more lineages left when the recombination event occurs, the larger effect it has, implying that it is mainly young recombination events that we detect when estimating the rate. If the population is growing, though, more lineages are present back in time and relatively more ancient recombination events...

  7. Polarization of the dielectronic recombination satellites including the generalized Breit interaction effects

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The polarizations of dielectronic recombination satellite lines for hydrogenlike F to U ions are calculated including the generalized Breit interaction(GBI). The calculated values of the asymmetry parameter are compared with other theoretical results and good agreements are obtained for all lines except for those from the level 2s2p3P1. The GBI effects on the polarization become much stronger as the atomic number increases. However, for different lines, the GBI effects on the polarization are different.

  8. [Recombination frequency in the locus Gli-D1 of common wheat T. aestivum L].

    Science.gov (United States)

    Kozub, N A; Sozinov, I A; Sozinov, A A

    2003-01-01

    The recombination frequency at the gliadin locus Gli-D1 of common wheat was determined by the maximum likelihood method. Recombination was observed between the gene encoding the fastest omega-component of the allele Gli-D1j, and the genes encoding the other omega-gliadins of this allele. The frequency of recombination was 0.65 +/- 0.18% for the cross between the near-isogenic lines of winter common wheat with respect to gliadin loci Gli-D1-4 and Gli-B1-3 and 0.78 +/- 0.45% for the cross between the varieties Yunnat and B-16.

  9. Regulation of Rad51-Mediated Homologous Recombination by BRCA2, DSS1 and RAD52

    DEFF Research Database (Denmark)

    Rants, Louise Olthaver Juhl

    Homologous recombination (HR) provides a mechanism to restore integrity and maintain stability of the genetic material. HR is a major pathway for repair of DNA double-strand breaks (DSB), recovery of broken replication forks and generation of meiotic crossovers. The defining step in HR...... in governing the activity of Rad51 and to learn how other recombination-associated proteins such as DSS1 and RAD52 contribute to its regulation. We use the yeast-like fungus Ustilago maydis and the avian DT40 cell line as experimental systems since both have a well-conserved BRCA2-based recombinational repair...

  10. A multi-omics analysis of recombinant protein production in Hek293 cells.

    Directory of Open Access Journals (Sweden)

    Stefanie Dietmair

    Full Text Available Hek293 cells are the predominant hosts for transient expression of recombinant proteins and are used for stable expression of proteins where post-translational modifications performed by CHO cells are inadequate. Nevertheless, there is little information available on the key cellular features underpinning recombinant protein production in Hek293 cells. To improve our understanding of recombinant protein production in Hek293 cells and identify targets for the engineering of an improved host cell line, we have compared a stable, recombinant protein producing Hek293 cell line and its parental cell line using a combination of transcriptomics, metabolomics and fluxomics. Producer cultures consumed less glucose than non-producer cultures while achieving the same growth rate, despite the additional burden of recombinant protein production. Surprisingly, there was no indication that producer cultures compensated for the reduction in glycolytic energy by increasing the efficiency of glucose utilization or increasing glutamine consumption. In contrast, glutamine consumption was lower and the majority of genes involved in oxidative phosphorylation were downregulated in producer cultures. We observed an overall downregulation of a large number of genes associated with broad cellular functions (e.g., cell growth and proliferation in producer cultures, and therefore speculate that a broad adaptation of the cellular network freed up resources for recombinant protein production while maintaining the same growth rate. Increased abundance of genes associated with endoplasmic reticulum stress indicated a possible bottleneck at the point of protein folding and assembly.

  11. A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells

    Science.gov (United States)

    Dietmair, Stefanie; Hodson, Mark P.; Quek, Lake-Ee; Timmins, Nicholas E.; Gray, Peter; Nielsen, Lars K.

    2012-01-01

    Hek293 cells are the predominant hosts for transient expression of recombinant proteins and are used for stable expression of proteins where post-translational modifications performed by CHO cells are inadequate. Nevertheless, there is little information available on the key cellular features underpinning recombinant protein production in Hek293 cells. To improve our understanding of recombinant protein production in Hek293 cells and identify targets for the engineering of an improved host cell line, we have compared a stable, recombinant protein producing Hek293 cell line and its parental cell line using a combination of transcriptomics, metabolomics and fluxomics. Producer cultures consumed less glucose than non-producer cultures while achieving the same growth rate, despite the additional burden of recombinant protein production. Surprisingly, there was no indication that producer cultures compensated for the reduction in glycolytic energy by increasing the efficiency of glucose utilization or increasing glutamine consumption. In contrast, glutamine consumption was lower and the majority of genes involved in oxidative phosphorylation were downregulated in producer cultures. We observed an overall downregulation of a large number of genes associated with broad cellular functions (e.g., cell growth and proliferation) in producer cultures, and therefore speculate that a broad adaptation of the cellular network freed up resources for recombinant protein production while maintaining the same growth rate. Increased abundance of genes associated with endoplasmic reticulum stress indicated a possible bottleneck at the point of protein folding and assembly. PMID:22937046

  12. Embryogenesis in Culture of Isolated Microspores of Brassica juncea var.capitata%结球芥菜游离小孢子培养的胚状体发生

    Institute of Scientific and Technical Information of China (English)

    王长龙; 杨培新; 郑奕雄

    2011-01-01

    2009~2011年对6种不同结球芥菜品种进行了小孢子培养试验,结果表明:在相同的培养条件下,各品种胚状体的发育是不同步的,胚状体的平均产量也存在极显著的差异,以大平铺的平均产胚量最高,为3.52个/蕾,其次为哥历蕾.%The isolated microspores of 6 Brassica juncea var. Capitata varieties were cultured from 2009 to 2011. The results showed that: under the same culture conditions, the embryogenesis of different varieties was of asynchrony, the average embryoid yield had extremely significant difference among different varieties, and the variety Dapingpu had the highest average embryoid yield (3.52 embryoids per bud) , followed by Gelilei.

  13. Punctuated distribution of recombination hotspots and demarcation of pericentromeric regions in Phaseolus vulgaris L.

    Science.gov (United States)

    Bhakta, Mehul S; Jones, Valerie A; Vallejos, C Eduardo

    2015-01-01

    High density genetic maps are a reliable tool for genetic dissection of complex plant traits. Mapping resolution is often hampered by the variable crossover and non-crossover events occurring across the genome, with pericentromeric regions (pCENR) showing highly suppressed recombination rates. The efficiency of linkage mapping can further be improved by characterizing and understanding the distribution of recombinational activity along individual chromosomes. In order to evaluate the genome wide recombination rate in common beans (Phaseolus vulgaris L.) we developed a SNP-based linkage map using the genotype-by-sequencing approach with a 188 recombinant inbred line family generated from an inter gene pool cross (Andean x Mesoamerican). We identified 1,112 SNPs that were subsequently used to construct a robust linkage map with 11 groups, comprising 513 recombinationally unique marker loci spanning 943 cM (LOD 3.0). Comparative analysis showed that the linkage map spanned >95% of the physical map, indicating that the map is almost saturated. Evaluation of genome-wide recombination rate indicated that at least 45% of the genome is highly recombinationally suppressed, and allowed us to estimate locations of pCENRs. We observed an average recombination rate of 0.25 cM/Mb in pCENRs as compared to the rest of genome that showed 3.72 cM/Mb. However, several hot spots of recombination were also detected with recombination rates reaching as high as 34 cM/Mb. Hotspots were mostly found towards the end of chromosomes, which also happened to be gene-rich regions. Analyzing relationships between linkage and physical map indicated a punctuated distribution of recombinational hot spots across the genome.

  14. Rad51 and Rad52 are involved in homologous recombination of replicating herpes simplex virus DNA.

    Directory of Open Access Journals (Sweden)

    Ka-Wei Tang

    Full Text Available Replication of herpes simplex virus 1 is coupled to recombination, but the molecular mechanisms underlying this process are poorly characterized. The role of Rad51 and Rad52 recombinases in viral recombination was examined in human fibroblast cells 1BR.3.N (wild type and in GM16097 with replication defects caused by mutations in DNA ligase I. Intermolecular recombination between viruses, tsS and tsK, harboring genetic markers gave rise to ∼17% recombinants in both cell lines. Knock-down of Rad51 and Rad52 by siRNA reduced production of recombinants to 11% and 5%, respectively, in wild type cells and to 3% and 5%, respectively, in GM16097 cells. The results indicate a specific role for Rad51 and Rad52 in recombination of replicating herpes simplex virus 1 DNA. Mixed infections using clinical isolates with restriction enzyme polymorphisms in the US4 and US7 genes revealed recombination frequencies of 0.7%/kbp in wild type cells and 4%/kbp in GM16097 cells. Finally, tandem repeats in the US7 gene remained stable upon serial passage, indicating a high fidelity of recombination in infected cells.

  15. 大豆重组自交家系群体动态株高及其相对生长速率与产量的关系%Relationship of Dynamic Plant Height and Its Relative Growth Rate with Yield Using Recombinant Inbred Lines of Soybean

    Institute of Scientific and Technical Information of China (English)

    黄中文; 王伟; 徐新娟; 文自翔; 李海朝; 李金英; 卢为国

    2011-01-01

    Selection for increasing yield potential is consistantly the main goal of soybean breeding. Plant height influences yield with uncertainty in soybean. A population of 212 recombinant inbred lines (RIL) was used to analyze the correlation of dynamic plant height and its relative growth rate with yield across two-year field experiments. The results were as followed: (1) The range of yield and plant height was 1 000-5 000 kg ha-1, 38-128 cm respectively, which indicated the sample in the study had a good representativeness. Significant differences between plant height and its relative growth rate at the same developmental stages were observed among the RILs. (2) Yield was significantly and positively correlated with plant height at 20 days after emergence, and the correlation increased with the growing stages and the highest correlation occurred when plant height culminated. Yield was positively correlated with relative growth rate in plant height at earlier growth stages and negatively correlated at later stages,indicating that faster growth in plant height at later stage would not be beneficial to higher yield. (3) A negative exponential regression mode was observed in the relationship between yield and plant height. Yield increased steeply when plant height was less than 80 cm and this trend would slow down when plant height was over 80 cm. We found yields reached 4 000 kg ha-1 when plant heights were 70-90 cm. The merits and demerits of phenotypic analysis using RIL and how to select plant height in soybean breeding were discussed.%提高产量潜力始终是大豆育种的重要目标,研究产量相关性状是解析产量重要途径之一.大豆株高影响产量,但存在不确定性.本研究利用一套包含212个家系的大豆重组自交家系(RIL),于2008-2009年连续2年测定各家系动态株高,并计算相对生长速率,研究与产量的关系,以期为大豆产量改良中对株高的选择提供参考信息.试验结果表明:(1)产

  16. 利用青海大黄油菜与黑芥人工合成芥菜型油菜%New Brassica juncea Sythesized from Interspecific Hybridization Between B. rapa (Qinghai dahuang) and B. nigra

    Institute of Scientific and Technical Information of China (English)

    赵志刚

    2014-01-01

    Using Brassica rapa and Brassica nigra as materials, new Brassic juncea can be synthesized through interspecific hybridization between B. rapa and B. nigra . The hybrid plants of B. rapa×B. nigra were produced through embryo culture and induced to shoot on culture MS medium. The hybrid plants had an intermediate morphology. Cytological analysis showed that the somatic of F1 plants had 18 chromosomes, which is the sum of gametic chromosomes of female parent B. campestris (2n=AA=20) and male parent B. nigra (2n=BB=16). SSR molecular markers analysis showed that these obtained plants were true interspecific hybrid between B. rapa (Qinghai dahuang) and B. nigra. The results will provide a new method for innovating germplasm resource of Brassiac juncea.%以芸薹属植物青海大黄油菜(Brassica rape)和黑芥(Brassica nigra)为研究对象,对其常规杂交后,通过离体胚培养获得的杂种种子经MS培养基诱导培养成苗,获得了青海大黄油菜与黑芥的杂种F1代植株,形态上表现为中间类型。通过细胞学的方法,鉴定了杂种F1代植株的细胞染色体数为18条,为母本青海大黄油菜(2=AA=20)和父本黑芥(2n=BB=16)的配子染色体数之和;SSR分子鉴定进一步表明,该杂种植株为真杂种。

  17. GARD: a genetic algorithm for recombination detection

    National Research Council Canada - National Science Library

    Kosakovsky Pond, Sergei L; Posada, David; Gravenor, Michael B; Woelk, Christopher H; Frost, Simon D W

    2006-01-01

    .... We developed a likelihood-based model selection procedure that uses a genetic algorithm to search multiple sequence alignments for evidence of recombination breakpoints and identify putative recombinant sequences...

  18. Initiation of meiotic recombination in Ustilago maydis

    National Research Council Canada - National Science Library

    Kojic, Milorad; Sutherland, Jeanette H; Pérez-Martín, José; Holloman, William K

    2013-01-01

    .... Ustilago maydis, a biotrophic fungus that parasitizes maize, has long been utilized as an experimental system for studying recombination, but it has not been clear when in the life cycle meiotic recombination initiates. U...

  19. Recombinant DNA: History of the Controversy.

    Science.gov (United States)

    Vigue, Charles L.; Stanziale, William G.

    1979-01-01

    The hazards associated with recombinant DNA research are presented along with some social implications and the development of recombinant DNA research guidelines by the National Institutes of Health. (SA)

  20. Homology requirements for recombination in Escherichia coli.

    OpenAIRE

    Watt, V M; Ingles, C J; Urdea, M S; Rutter, W J

    1985-01-01

    The DNA sequence homology required for recombination in Escherichia coli has been determined by measuring the recombination frequency between insulin DNA in a miniplasmid pi VX and a homologous sequence in a bacteriophage lambda vector. A minimum of approximately equal to 20 base pairs in a completely homologous segment is required for significant recombination. There is an exponential increase in the frequency of recombination when the length of homologous DNA is increased from 20 base pairs...

  1. Determination of Interactions Between the Two Determinant Transcription Factors of Flowering Signal Integrators in Vitro in Brassica juncea Coss.(Mustard)%芥菜开花信号整合子的两个核心转录因子FLC和SVP相互作用的体外检测

    Institute of Scientific and Technical Information of China (English)

    汤青林; 许俊强; 宋明; 王志敏

    2011-01-01

    为了深入研究调控芥菜开花信号整合子的两个核心转录因子Flowering Locus C(FLC)和SHOTR VEGETATIVE PHASE(SVP)的作用机理和进行人工调控,以芥菜‘QJ’为材料,采用RT-PCR技术分别扩增FLC和SVP的编码序列,构建原核表达质粒pET43.1a-FLC、pET43.1a-SVP,转化宿主菌大肠杆菌BL21,通过SDS-PAGE检测该蛋白的表达。利用免疫共沉淀原理及pET43.1a-FLC、pET43.1a-SVP融合蛋白序列中的6×His标签能与Ni+结合的特点,对芥菜FLC与SVP的相互作用进行SDS-PAGE检测。结果表明:FLC与SVP能在体外相互作用并形成复合体,为深入分析FLC与SVP间的作用机理以及探讨其与下游开花信号整合子元件的相互作用提供了理论和技术基础。%The fate of the flowering signal integrators is determined by Flowering Locus C(FLC) and SHOTR VEGETATIVE PHASE(SVP),which probably directly interact with each other to regulate flowering.For further study on mechanism and reagent control of the interactions between the two transcription factors,the protein interactions in vitro were testified in Brassica juncea Coss.(mustard) variety‘QJ’.In the present study,the coding sequences of FLC and SVP were amplified,and then inserted into the expression vector pET43.1a to construct the recombinant plasmids pET43.1a-FLC and pET43.1a-SVP.After transformation to E.coli(BL21),the expression of recombinant proteins were detected via SDS-PAGE.With co-immunoprecipitation and characteristic of 6 × His tag in fusion protein of pET43.1a-FLC and pET43.1a-SVP which could combine with Ni+,a new method was put forward to detecting the interactions between FLC and SVP proteins.Using this method,the interactions were analyzed via SDS-PAGE.The results showed that FLC and SVP could act with each other to combine and form a complex.This research provides theoretical and technical bases for analyzing the mechanism of interactions between FLC and SVP,for further

  2. Detection of the excited carbon lines at decameter waves in some galactic objects

    Energy Technology Data Exchange (ETDEWEB)

    Konovalenko, A.A.

    1984-12-01

    The absorption lines C638..cap alpha..-C641..cap alpha.. are detected at 25 MHz in the region with galactic coordinates l/sub 2/=75 deg; b/sub 2/=0 deg and near the NGC 2024 nebulae. The observed parameters of lines can be only explained in the frame of the low-temperature dielectronic recombination mechanism. The advantages of decametric observations of carbon recombination lines in the various components of the interstellar medium are pointed out.

  3. Single-crossover recombination and ancestral recombination trees.

    Science.gov (United States)

    Baake, Ellen; von Wangenheim, Ute

    2014-05-01

    We consider the Wright-Fisher model for a population of [Formula: see text] individuals, each identified with a sequence of a finite number of sites, and single-crossover recombination between them. We trace back the ancestry of single individuals from the present population. In the [Formula: see text] limit without rescaling of parameters or time, this ancestral process is described by a random tree, whose branching events correspond to the splitting of the sequence due to recombination. With the help of a decomposition of the trees into subtrees, we calculate the probabilities of the topologies of the ancestral trees. At the same time, these probabilities lead to a semi-explicit solution of the deterministic single-crossover equation. The latter is a discrete-time dynamical system that emerges from the Wright-Fisher model via a law of large numbers and has been waiting for a solution for many decades.

  4. Extended recombinant bacterial ghost system.

    Science.gov (United States)

    Lubitz, W; Witte, A; Eko, F O; Kamal, M; Jechlinger, W; Brand, E; Marchart, J; Haidinger, W; Huter, V; Felnerova, D; Stralis-Alves, N; Lechleitner, S; Melzer, H; Szostak, M P; Resch, S; Mader, H; Kuen, B; Mayr, B; Mayrhofer, P; Geretschläger, R; Haslberger, A; Hensel, A

    1999-08-20

    Controlled expression of cloned PhiX174 gene E in Gram-negative bacteria results in lysis of the bacteria by formation of an E-specific transmembrane tunnel structure built through the cell envelope complex. Bacterial ghosts from a variety of bacteria are used as non-living candidate vaccines. In the recombinant ghost system, foreign proteins are attached on the inside of the inner membrane as fusions with specific anchor sequences. Ghosts have a sealed periplasmic space and the export of proteins into this space vastly extends the capacity of ghosts or recombinant ghosts to function as carriers of foreign antigens. In addition, S-layer proteins forming shell-like self assembly structures can be expressed in candidate vaccine strains prior to E-mediated lysis. Such recombinant S-layer proteins carrying foreign epitopes further extend the possibilities of ghosts as carriers of foreign epitopes. As ghosts have inherent adjuvant properties, they can be used as adjuvants in combination with subunit vaccines. Subunits or other ligands can also be coupled to matrixes like dextran which are used to fill the internal lumen of ghosts. Oral, aerogenic or parent