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Sample records for picea glauca bac

  1. Targeted isolation, sequence assembly and characterization of two white spruce (Picea glauca BAC clones for terpenoid synthase and cytochrome P450 genes involved in conifer defence reveal insights into a conifer genome

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    Ritland Carol

    2009-08-01

    Full Text Available Abstract Background Conifers are a large group of gymnosperm trees which are separated from the angiosperms by more than 300 million years of independent evolution. Conifer genomes are extremely large and contain considerable amounts of repetitive DNA. Currently, conifer sequence resources exist predominantly as expressed sequence tags (ESTs and full-length (FLcDNAs. There is no genome sequence available for a conifer or any other gymnosperm. Conifer defence-related genes often group into large families with closely related members. The goals of this study are to assess the feasibility of targeted isolation and sequence assembly of conifer BAC clones containing specific genes from two large gene families, and to characterize large segments of genomic DNA sequence for the first time from a conifer. Results We used a PCR-based approach to identify BAC clones for two target genes, a terpene synthase (3-carene synthase; 3CAR and a cytochrome P450 (CYP720B4 from a non-arrayed genomic BAC library of white spruce (Picea glauca. Shotgun genomic fragments isolated from the BAC clones were sequenced to a depth of 15.6- and 16.0-fold coverage, respectively. Assembly and manual curation yielded sequence scaffolds of 172 kbp (3CAR and 94 kbp (CYP720B4 long. Inspection of the genomic sequences revealed the intron-exon structures, the putative promoter regions and putative cis-regulatory elements of these genes. Sequences related to transposable elements (TEs, high complexity repeats and simple repeats were prevalent and comprised approximately 40% of the sequenced genomic DNA. An in silico simulation of the effect of sequencing depth on the quality of the sequence assembly provides direction for future efforts of conifer genome sequencing. Conclusion We report the first targeted cloning, sequencing, assembly, and annotation of large segments of genomic DNA from a conifer. We demonstrate that genomic BAC clones for individual members of multi-member gene

  2. Targeted isolation, sequence assembly and characterization of two white spruce (Picea glauca) BAC clones for terpenoid synthase and cytochrome P450 genes involved in conifer defence reveal insights into a conifer genome.

    Science.gov (United States)

    Hamberger, Björn; Hall, Dawn; Yuen, Mack; Oddy, Claire; Hamberger, Britta; Keeling, Christopher I; Ritland, Carol; Ritland, Kermit; Bohlmann, Jörg

    2009-08-06

    Conifers are a large group of gymnosperm trees which are separated from the angiosperms by more than 300 million years of independent evolution. Conifer genomes are extremely large and contain considerable amounts of repetitive DNA. Currently, conifer sequence resources exist predominantly as expressed sequence tags (ESTs) and full-length (FL)cDNAs. There is no genome sequence available for a conifer or any other gymnosperm. Conifer defence-related genes often group into large families with closely related members. The goals of this study are to assess the feasibility of targeted isolation and sequence assembly of conifer BAC clones containing specific genes from two large gene families, and to characterize large segments of genomic DNA sequence for the first time from a conifer. We used a PCR-based approach to identify BAC clones for two target genes, a terpene synthase (3-carene synthase; 3CAR) and a cytochrome P450 (CYP720B4) from a non-arrayed genomic BAC library of white spruce (Picea glauca). Shotgun genomic fragments isolated from the BAC clones were sequenced to a depth of 15.6- and 16.0-fold coverage, respectively. Assembly and manual curation yielded sequence scaffolds of 172 kbp (3CAR) and 94 kbp (CYP720B4) long. Inspection of the genomic sequences revealed the intron-exon structures, the putative promoter regions and putative cis-regulatory elements of these genes. Sequences related to transposable elements (TEs), high complexity repeats and simple repeats were prevalent and comprised approximately 40% of the sequenced genomic DNA. An in silico simulation of the effect of sequencing depth on the quality of the sequence assembly provides direction for future efforts of conifer genome sequencing. We report the first targeted cloning, sequencing, assembly, and annotation of large segments of genomic DNA from a conifer. We demonstrate that genomic BAC clones for individual members of multi-member gene families can be isolated in a gene-specific fashion. The

  3. Modular organization of the white spruce (Picea glauca) transcriptome reveals functional organization and evolutionary signatures.

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    Raherison, Elie S M; Giguère, Isabelle; Caron, Sébastien; Lamara, Mebarek; MacKay, John J

    2015-07-01

    Transcript profiling has shown the molecular bases of several biological processes in plants but few studies have developed an understanding of overall transcriptome variation. We investigated transcriptome structure in white spruce (Picea glauca), aiming to delineate its modular organization and associated functional and evolutionary attributes. Microarray analyses were used to: identify and functionally characterize groups of co-expressed genes; investigate expressional and functional diversity of vascular tissue preferential genes which were conserved among Picea species, and identify expression networks underlying wood formation. We classified 22 857 genes as variable (79%; 22 coexpression groups) or invariant (21%) by profiling across several vegetative tissues. Modular organization and complex transcriptome restructuring among vascular tissue preferential genes was revealed by their assignment to coexpression groups with partially overlapping profiles and partially distinct functions. Integrated analyses of tissue-based and temporally variable profiles identified secondary xylem gene networks, showed their remodelling over a growing season and identified PgNAC-7 (no apical meristerm (NAM), Arabidopsis transcription activation factor (ATAF) and cup-shaped cotyledon (CUC) transcription factor 007 in Picea glauca) as a major hub gene specific to earlywood formation. Reference profiling identified comprehensive, statistically robust coexpressed groups, revealing that modular organization underpins the evolutionary conservation of the transcriptome structure. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  4. Effects of cloning and root-tip size on observations of fungal ITS sequences from Picea glauca roots

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    Daniel L. Lindner; Mark T. Banik

    2009-01-01

    To better understand the effects of cloning on observations of fungal ITS sequences from Picea glauca (white spruce) roots two techniques were compared: (i) direct sequencing of fungal ITS regions from individual root tips without cloning and (ii) cloning and sequencing of fungal ITS regions from individual root tips. Effect of root tip size was...

  5. Improvement of grafting procedures for the ornmental species: I. Picea pungens Engelm. var. glauca Regel

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    Ioan Blada

    2011-11-01

    Full Text Available In order to get ornamental trees for landscaping, Colorado blue spuce (Picea pungens Engelm. var. glauca Regel scions were grafted on Norway spruce (Picea abies (L. Karst. rootstocks.An original double-side-veneer grafting method was applied in five experimental or basic variants. The plastic tapes and the ecological Ceraltin® wax developed by the Research and Development of Bio-stimulators (CCDB BIOS Cluj were tested. In addition, two controls in which the classic raffia and the traditional hot wax and the classic side-veneer-grafting method were used. The obtained results, expressed in percents, were transformed in arcsin square root of percent values, and then a two-way analysis of variance (ANOVA was used. Highly significant (p < 0.001 statistical differences were found between all grafting variants, including controls. The Duncan test proved that original double-side-veneer grafting method and the new developed materials, i.e. plastic tapes and the ecological Ceraltin® wax have contributed in getting a better grafting success compared to the controls.Thus, the double-side-veneer grafting method and the two grafting materials are highly recommended to be used in getting grafted Colorado blue spuce ornamental trees.

  6. Improvement of grafting procedures for the ornamental species: I. Picea pungens Engelm. var. glauca Regel

    Directory of Open Access Journals (Sweden)

    Ioan Blada

    2013-12-01

    Full Text Available In order to get ornamental trees for landscaping, Colorado blue spuce (Picea pungens Engelm. var.glauca Regel scions were grafted on Norway spruce (Picea abies (L. Karst. rootstocks. An original double-side-veneer grafting method was applied in five experimental or basic variants. The plastic tapes and the ecological Ceraltin wax developed by the Research and Development of Bio-stimulators (CCDB BIOS Cluj were tested. In addition, two controls in which the classic raffia and the traditional hot wax and the classic side-veneer-grafting method were used. The obtained results, expressed in percents, were transformed in arcsin square root of percent values, and then a two-way analysis of variance (ANOVA was used. Highly significant (p < 0.001 statistical differences were found between all grafting variants, including controls. The Duncan test proved that original double-side-veneer grafting method and the new developed materials, i.e. plastic tapes and the ecological Ceraltin wax have contributed in getting a better grafting success compared to the controls.Thus, the double-side-veneer grafting method and the two grafting materials are highly recommended to be used in getting grafted Colorado blue spuce ornamental trees.

  7. Overexpression of Laccaria bicolor aquaporin JQ585595 alters root water transport properties in ectomycorrhizal white spruce (Picea glauca) seedlings.

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    Xu, Hao; Kemppainen, Minna; El Kayal, Walid; Lee, Seong Hee; Pardo, Alejandro G; Cooke, Janice E K; Zwiazek, Janusz J

    2015-01-01

    The contribution of hyphae to water transport in ectomycorrhizal (ECM) white spruce (Picea glauca) seedlings was examined by altering expression of a major water-transporting aquaporin in Laccaria bicolor. Picea glauca was inoculated with wild-type (WT), mock transgenic or L. bicolor aquaporin JQ585595-overexpressing (OE) strains and exposed to root temperatures ranging from 5 to 20°C to examine the root water transport properties, physiological responses and plasma membrane intrinsic protein (PIP) expression in colonized plants. Mycorrhization increased shoot water potential, transpiration, net photosynthetic rates, root hydraulic conductivity and root cortical cell hydraulic conductivity in seedlings. At 20°C, OE plants had higher root hydraulic conductivity compared with WT plants and the increases were accompanied by higher expression of P. glauca PIP GQ03401_M18.1 in roots. In contrast to WT L. bicolor, the effects of OE fungi on root and root cortical cell hydraulic conductivities were abolished at 10 and 5°C in the absence of major changes in the examined transcript levels of P. glauca root PIPs. The results provide evidence for the importance of fungal aquaporins in root water transport of mycorrhizal plants. They also demonstrate links between hyphal water transport, root aquaporin expression and root water transport in ECM plants. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  8. Single-Step BLUP with Varying Genotyping Effort in Open-Pollinated Picea glauca.

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    Ratcliffe, Blaise; El-Dien, Omnia Gamal; Cappa, Eduardo P; Porth, Ilga; Klápště, Jaroslav; Chen, Charles; El-Kassaby, Yousry A

    2017-03-10

    Maximization of genetic gain in forest tree breeding programs is contingent on the accuracy of the predicted breeding values and precision of the estimated genetic parameters. We investigated the effect of the combined use of contemporary pedigree information and genomic relatedness estimates on the accuracy of predicted breeding values and precision of estimated genetic parameters, as well as rankings of selection candidates, using single-step genomic evaluation (HBLUP). In this study, two traits with diverse heritabilities [tree height (HT) and wood density (WD)] were assessed at various levels of family genotyping efforts (0, 25, 50, 75, and 100%) from a population of white spruce ( Picea glauca ) consisting of 1694 trees from 214 open-pollinated families, representing 43 provenances in Québec, Canada. The results revealed that HBLUP bivariate analysis is effective in reducing the known bias in heritability estimates of open-pollinated populations, as it exposes hidden relatedness, potential pedigree errors, and inbreeding. The addition of genomic information in the analysis considerably improved the accuracy in breeding value estimates by accounting for both Mendelian sampling and historical coancestry that were not captured by the contemporary pedigree alone. Increasing family genotyping efforts were associated with continuous improvement in model fit, precision of genetic parameters, and breeding value accuracy. Yet, improvements were observed even at minimal genotyping effort, indicating that even modest genotyping effort is effective in improving genetic evaluation. The combined utilization of both pedigree and genomic information may be a cost-effective approach to increase the accuracy of breeding values in forest tree breeding programs where shallow pedigrees and large testing populations are the norm. Copyright © 2017 Ratcliffe et al.

  9. Single-Step BLUP with Varying Genotyping Effort in Open-Pollinated Picea glauca

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    Blaise Ratcliffe

    2017-03-01

    Full Text Available Maximization of genetic gain in forest tree breeding programs is contingent on the accuracy of the predicted breeding values and precision of the estimated genetic parameters. We investigated the effect of the combined use of contemporary pedigree information and genomic relatedness estimates on the accuracy of predicted breeding values and precision of estimated genetic parameters, as well as rankings of selection candidates, using single-step genomic evaluation (HBLUP. In this study, two traits with diverse heritabilities [tree height (HT and wood density (WD] were assessed at various levels of family genotyping efforts (0, 25, 50, 75, and 100% from a population of white spruce (Picea glauca consisting of 1694 trees from 214 open-pollinated families, representing 43 provenances in Québec, Canada. The results revealed that HBLUP bivariate analysis is effective in reducing the known bias in heritability estimates of open-pollinated populations, as it exposes hidden relatedness, potential pedigree errors, and inbreeding. The addition of genomic information in the analysis considerably improved the accuracy in breeding value estimates by accounting for both Mendelian sampling and historical coancestry that were not captured by the contemporary pedigree alone. Increasing family genotyping efforts were associated with continuous improvement in model fit, precision of genetic parameters, and breeding value accuracy. Yet, improvements were observed even at minimal genotyping effort, indicating that even modest genotyping effort is effective in improving genetic evaluation. The combined utilization of both pedigree and genomic information may be a cost-effective approach to increase the accuracy of breeding values in forest tree breeding programs where shallow pedigrees and large testing populations are the norm.

  10. Modeling the distribution of white spruce (Picea glauca) for Alaska with high accuracy: an open access role-model for predicting tree species in last remaining wilderness areas

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    Bettina Ohse; Falk Huettmann; Stefanie M. Ickert-Bond; Glenn P. Juday

    2009-01-01

    Most wilderness areas still lack accurate distribution information on tree species. We met this need with a predictive GIS modeling approach, using freely available digital data and computer programs to efficiently obtain high-quality species distribution maps. Here we present a digital map with the predicted distribution of white spruce (Picea glauca...

  11. Conservation and divergence of gene expression plasticity following c. 140 million years of evolution in lodgepole pine (Pinus contorta) and interior spruce (Picea glauca×Picea engelmannii).

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    Yeaman, Sam; Hodgins, Kathryn A; Suren, Haktan; Nurkowski, Kristin A; Rieseberg, Loren H; Holliday, Jason A; Aitken, Sally N

    2014-07-01

    Species respond to environmental stress through a combination of genetic adaptation and phenotypic plasticity, both of which may be important for survival in the face of climatic change. By characterizing the molecular basis of plastic responses and comparing patterns among species, it is possible to identify how such traits evolve. Here, we used de novo transcriptome assembly and RNAseq to explore how patterns of gene expression differ in response to temperature, moisture, and light regime treatments in lodgepole pine (Pinus contorta) and interior spruce (a natural hybrid population of Picea glauca and Picea engelmannii). We found wide evidence for an effect of treatment on expression within each species, with 6413 and 11,658 differentially expressed genes identified in spruce and pine, respectively. Comparing patterns of expression among these species, we found that 74% of all orthologs with differential expression had a pattern that was conserved in both species, despite 140 million yr of evolution. We also found that the specific treatments driving expression patterns differed between genes with conserved versus diverged patterns of expression. We conclude that natural selection has probably played a role in shaping plastic responses to environment in these species. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  12. A comparison of genomic selection models across time in interior spruce (Picea engelmannii × glauca) using unordered SNP imputation methods.

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    Ratcliffe, B; El-Dien, O G; Klápště, J; Porth, I; Chen, C; Jaquish, B; El-Kassaby, Y A

    2015-12-01

    Genomic selection (GS) potentially offers an unparalleled advantage over traditional pedigree-based selection (TS) methods by reducing the time commitment required to carry out a single cycle of tree improvement. This quality is particularly appealing to tree breeders, where lengthy improvement cycles are the norm. We explored the prospect of implementing GS for interior spruce (Picea engelmannii × glauca) utilizing a genotyped population of 769 trees belonging to 25 open-pollinated families. A series of repeated tree height measurements through ages 3-40 years permitted the testing of GS methods temporally. The genotyping-by-sequencing (GBS) platform was used for single nucleotide polymorphism (SNP) discovery in conjunction with three unordered imputation methods applied to a data set with 60% missing information. Further, three diverse GS models were evaluated based on predictive accuracy (PA), and their marker effects. Moderate levels of PA (0.31-0.55) were observed and were of sufficient capacity to deliver improved selection response over TS. Additionally, PA varied substantially through time accordingly with spatial competition among trees. As expected, temporal PA was well correlated with age-age genetic correlation (r=0.99), and decreased substantially with increasing difference in age between the training and validation populations (0.04-0.47). Moreover, our imputation comparisons indicate that k-nearest neighbor and singular value decomposition yielded a greater number of SNPs and gave higher predictive accuracies than imputing with the mean. Furthermore, the ridge regression (rrBLUP) and BayesCπ (BCπ) models both yielded equal, and better PA than the generalized ridge regression heteroscedastic effect model for the traits evaluated.

  13. Clonal variation in gas exchange and freezing tolerance development of interior spruce (Picea glauca (Moench) Voss x P. engelmannii Parry ex Engelm.) during autumn acclimation

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    Fan, Shihe; Grossnickle, S.C. [British Columbia Research Corp., Vancouver, BC (Canada). Forest Biotechnology Centre

    1999-08-01

    Variation in physiological response during autumn acclimation was investigated in somatic seedlings of 10 interior spruce (Picea glauca (Moench) Voss x P. engelmannii Parry ex Engelm.) clones from five full-sib families. Experiments were conducted on 2-yr-old seedlings through simulation in a growth chamber. Throughout the experimental period, gas-exchange parameters (net photosynthesis, stomatal conductance to water vapour and instantaneous water-use efficiency) were measured weekly and freezing tolerance was determined five times. Results showed that as seedlings acclimated to decreasing air temperature and photoperiod, stomatal conductance decreased linearly, photosynthesis was unchanged until air temperature and photoperiod were below 10 deg C and 11 h, respectively, water-use efficiency nearly doubled and freezing tolerance increased in a curvilinear fashion. There was significant between- and within-family clonal variation in all of these physiological parameters. 47 refs, 7 figs

  14. Habitat conditions and phenological tree traits overrule the influence of tree genotype in the needle mycobiome-Picea glauca system at an arctic treeline ecotone.

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    Eusemann, Pascal; Schnittler, Martin; Nilsson, R Henrik; Jumpponen, Ari; Dahl, Mathilde B; Würth, David G; Buras, Allan; Wilmking, Martin; Unterseher, Martin

    2016-09-01

    Plant-associated mycobiomes in extreme habitats are understudied and poorly understood. We analysed Illumina-generated ITS1 sequences from the needle mycobiome of white spruce (Picea glauca) at the northern treeline in Alaska (USA). Sequences were obtained from the same DNA that was used for tree genotyping. In the present study, fungal metabarcoding and tree microsatellite data were compared for the first time. In general, neighbouring trees shared more fungal taxa with each other than trees growing in further distance. Mycobiomes correlated strongly with phenological host traits and local habitat characteristics contrasting a dense forest stand with an open treeline site. Genetic similarity between trees did not influence fungal composition and no significant correlation existed between needle mycobiome and tree genotype. Our results suggest the pronounced influence of local habitat conditions and phenotypic tree traits on needle-inhabiting fungi. By contrast, the tree genetic identity cannot be benchmarked as a dominant driver for needle-inhabiting mycobiomes, at least not for white spruce in this extreme environment. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  15. Variation in Wood Quality in White Spruce (Picea Glauca (Moench Voss. Part I. Defining the Juvenile–Mature Wood Transition Based on Tracheid Length

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    Cyriac Serge Mvolo

    2015-01-01

    Full Text Available Estimations of transition age (TA and juvenile wood proportion (JWP are important for wood industries due to their impact on end-product quality. However, the relationships between analytical determination of TA based on tracheid length (TL and recognized thresholds for adequate end products have not yet been established. In this study, we used three different statistical models to estimate TA in white spruce (Picea glauca (Moench Voss based on TL radial variation. We compared the results with technological maturity. A two-millimeter threshold, previously suggested for good paper tear strength, was used. Tracheid length increased from pith to bark and from breast height to upper height. Juvenile wood (JW was conical with the three models. At breast height, TA ranged from 11 to 27 years and JWP ranged from 15.3% to 47.5% across the three models. The linear mixed model produced more conservative estimates than the maximum-quadratic-linear (M_Q_L model. Both the linear mixed model and the M_Q_L model produced more conservative TA estimates than the piecewise model. TA estimates by the MIXED model, and to a lesser extent by the M_Q_L model, were equivalent to those for real mature wood, whereas TA estimates by the piecewise model were considerably lower, falling into the transition wood area.

  16. Conifer R2R3-MYB transcription factors: sequence analyses and gene expression in wood-forming tissues of white spruce (Picea glauca

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    Grima-Pettenati Jacqueline

    2007-03-01

    Full Text Available Abstract Background Several members of the R2R3-MYB family of transcription factors act as regulators of lignin and phenylpropanoid metabolism during wood formation in angiosperm and gymnosperm plants. The angiosperm Arabidopsis has over one hundred R2R3-MYBs genes; however, only a few members of this family have been discovered in gymnosperms. Results We isolated and characterised full-length cDNAs encoding R2R3-MYB genes from the gymnosperms white spruce, Picea glauca (13 sequences, and loblolly pine, Pinus taeda L. (five sequences. Sequence similarities and phylogenetic analyses placed the spruce and pine sequences in diverse subgroups of the large R2R3-MYB family, although several of the sequences clustered closely together. We searched the highly variable C-terminal region of diverse plant MYBs for conserved amino acid sequences and identified 20 motifs in the spruce MYBs, nine of which have not previously been reported and three of which are specific to conifers. The number and length of the introns in spruce MYB genes varied significantly, but their positions were well conserved relative to angiosperm MYB genes. Quantitative RTPCR of MYB genes transcript abundance in root and stem tissues revealed diverse expression patterns; three MYB genes were preferentially expressed in secondary xylem, whereas others were preferentially expressed in phloem or were ubiquitous. The MYB genes expressed in xylem, and three others, were up-regulated in the compression wood of leaning trees within 76 hours of induction. Conclusion Our survey of 18 conifer R2R3-MYB genes clearly showed a gene family structure similar to that of Arabidopsis. Three of the sequences are likely to play a role in lignin metabolism and/or wood formation in gymnosperm trees, including a close homolog of the loblolly pine PtMYB4, shown to regulate lignin biosynthesis in transgenic tobacco.

  17. Interspecific Competition and Trade-offs in Resource Allocation are the Key to Successful Growth of Seedlings of White Spruce (Picea glauca (Moench) Voss) at Subarctic Treelines in Warming Alaska.

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    Okano, K.; Bret-Harte, M. S.

    2015-12-01

    Alpine treelines in Alaska have advanced for the past 50 years in response to the recent climate warming. However, further increases in temperatures may cause treeline species drought stress and increase susceptibility to insect outbreaks and fire. Complex factors such as soil conditions and plant species composition also impact the growth of seedlings, which are essential to sustain boreal forests. Our goals were to assess 1) the current optimal elevation for the treeline species Picea glauca (white spruce) seedlings and how it is altered by climate change, and 2) their growth/survival strategies at each environmental site. We studied the growth response of spruce seedlings along an altitudinal gradient at 6 sites, consisting of tundra, forest, or transitional ecotone in Denali National Park and one forest site in Fairbanks, AK. In May 2012, four-month old seedlings were planted with or without naturally occurring plants to compare the presence or absence of the interspecific interaction. Summer temperatures were increased by one small greenhouse per site. Over 2 growing seasons, growth was measured non-destructively, and then the seedlings were harvested. Relative growth rate (RGR) in height was increased significantly as the altitude was increased. Elevated temperature increased height only in seedlings at a high-altitude forest. Seedlings with neighboring plants had a higher RGR in height than seedlings that had neighbors removed, while significantly wider diameters were measured from the seedlings without neighbors. A weak trend of declining diameter width with increasing altitudes was seen. Seedlings that grew taller did not grow their stems wider, indicating trade-offs in resource allocation. None of the altitudinal sites had a clear advantage for the growth of the seedlings. Habitat microclimate and the interaction with other species could be more important than the altitude or temperatures and hence, key to the survival and growth of spruce seedlings in

  18. SOMATIC MITOSES IN CELLS OF PICEA GLUCA CULTIVATED IN VITRO.

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    RISSER, P G

    1964-02-07

    The cytology of one strain of tumor tissue from the spruce tree, Picea glauca, grown in vitro for more than a year was examined. The cells of this strain are characterized by a uniform chromosome number of 24, and the strain appears to be quite stable. The implications of the results of this study and previous studies on similar material are discussed.

  19. Demonstration of paternal inheritance of plastids in Picea (Pinaceae)

    International Nuclear Information System (INIS)

    Stine, M.

    1988-01-01

    Chloroplast DNA (cpDNA) was purified from Picea glauca, P. pungens, P. engelmannii, and P. omorika, and was digested with several restriction endonucleases. Interspecific restriction fragment length polymorphisms (RFLPs) of cpDNA were identified. The RFLPs were identified as cpDNA by the hybridization of cloned, 32 -P labeled, petunia cpDNA to the polymorphic bands, and by the lack of hybridization of a cloned and labeled mtDNA probe from maize. Chloroplast DNA RFLPs that showed no intraspecific variation when examined across the natural range for each species, were used as markers to follow the inheritance of plastids in interspecific hybrids. The inheritance of plastids was determined for F 1 -hybrids from reciprocal crosses of P. glauca and P. pungens, P. glauca and P. omorika, and F 1 -hybrids of P. engelmannii x pungens. All 31 F 1 -hybrids examined showed the cpDNA genotypes of the pollen parent, or the paternal species

  20. Nicotiana glauca poisoning in ostriches (Struthio camelus)

    CSIR Research Space (South Africa)

    Botha, CJ

    2011-01-01

    Full Text Available Putative Nicotiana glauca (wild tobacco) poisoning was diagnosed in a flock of ostriches near Oudtshoorn, South Africa. Post mortem examinations (n = 7) were performed on ostriches (Struthio camelus) that had died. Suspicious leaf remnants (weighing...

  1. Recombining overlapping BACs into a single larger BAC

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    Huxley Clare

    2004-01-01

    Full Text Available Abstract Background BAC clones containing entire mammalian genes including all the transcribed region and long range controlling elements are very useful for functional analysis. Sequenced BACs are available for most of the human and mouse genomes and in many cases these contain intact genes. However, large genes often span more than one BAC, and single BACs covering the entire region of interest are not available. Here we describe a system for linking two or more overlapping BACs into a single clone by homologous recombination. Results The method was used to link a 61-kb insert carrying the final 5 exons of the human CFTR gene onto a 160-kb BAC carrying the first 22 exons. Two rounds of homologous recombination were carried out in the EL350 strain of bacteria which can be induced for the Red genes. In the first round, the inserts of the two overlapping BACs were subcloned into modified BAC vectors using homologous recombination. In the second round, the BAC to be added was linearised with the very rare-cutting enzyme I-PpoI and electroporated into recombination efficient EL350 bacteria carrying the other BAC. Recombined BACs were identified by antibiotic selection and PCR screening and 10% of clones contained the correctly recombined 220-kb BAC. Conclusion The system can be used to link the inserts from any overlapping BAC or PAC clones. The original orientation of the inserts is not important and desired regions of the inserts can be selected. The size limit for the fragments recombined may be larger than the 61 kb used here and multiple BACs in a contig could be combined by alternating use of the two pBACLink vectors. This system should be of use to many investigators wishing to carry out functional analysis on large mammalian genes which are not available in single BAC clones.

  2. Bac on the Border

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    Umberger, Emily

    2007-01-01

    Although now on the Tohono O'odom reservation in the modern US Southwest, when the Franciscan church of San Xavier del Bac was built (1780-97), its location was the northern frontier of New Spain. From the outset the church stood out from other northern New Spanish missions in its elaborate decoration, and it still stands out because its contents remain intact, despite changes through time. This essay serves as an introduction to the church as a subject of art historical study. It highlights ...

  3. Fight Bac! | Partnership for Food Safety Education

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    Fight Bac! Fight Bac! Fight Bac! Partnership for Food Safety Education Supporting consumers to & Symptoms Food Safety Glossary Food Safety Education Food Safety Education Month 2017 Don't Wing Spanish Resources Food Safety Education Food Safety Education Month 2017 Don't Wing It The Story of Your

  4. Molecular Variation in Picea Rubens and Picea Mariana

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    Bobola, Michael S.

    Restriction fragment length polymorphism (RFLP) variation was examined among samples from the entire range of red spruce (Picea rubens Sarg.), the eastern complex of black spruce (Picea mariana (Mill.) B.S.P.), control -cross red-on-black hybrids, and natural populations of red and black spruce. Within-species and population variation was examined. In addition an accurate species index capable of identifying red spruce, black spruce and hybridization between the two species was developed. The nuclear rDNA repeat unit size in Picea ranged from a minimum of 32 kbp to greater than 40 kbp, two to three fold larger than the typical angiosperm rDNA unit. At a size greater than 32 kbp and a concentration averaging 1.2-1.3 times 10^4 copies per pg genomic DNA, the rDNA repeat constitutes approximately 4% of the spruce genome. The rDNA repeat units were found to be polymorphic within an individual genome with up to five distinct rDNA repeat unit types (alleles) evident. The RFLPs observed in the rDNA repeat were not species specific; however, noticeable trends in internal allelic frequencies were noticed which were useful for between-species differentiation. One marker (EMW 4.35) displayed a significant relationship with geographic origins and habitat suggesting that the observed between-species variation for this marker may be due to selection rather than the result of a true species difference. Variation in the nuclear rDNA repeat could not accurately differentiate hybrids from black spruce. Additional markers were required to identify hybrids. RFLPs were identified for the organelle genomes of red spruce, and black spruce. The organelle inheritance pattern was deduced using controlled-cross hybrids. Organelle markers were combined with allelic data from the nuclear rDNA repeat to derive a simple three character index capable of identifying red spruce, black spruce and hybridization between the two species. Significant gene flow was observed between red and black spruce

  5. [Effects of simulating acid rain on photosynthesis and chlorophyll fluorescence parameters of Quercus glauca Quercus glauca].

    Science.gov (United States)

    Wang, Sai; Yi, Li-Ta; Yu, Shu-Quan; Zhang, Chao; Shi, Jing-Jing

    2014-08-01

    At three levels of simulated acid rainfall intensities with pH values of 2.5 (severe), 40 (medium) and 5.6 (light) respectively, the responses of chlorophyll fluorescence and photosynthetic parameters of Quercus glauca seedlings were studied in three acid rainfall treatments, i. e. only the aboveground of seedlings exposed to acid rain (T1), both of the seedlings and soil exposed to acid rain (T2), only the soil exposed to acid rain (T3) compared with blank control (CK). Under the severe acid rainfall, T1 significantly inhibited chlorophyll synthesis, and thus reduced the primary photochemical efficiency of PS II ( F(v)/F(m)), potential activity of PS II (F(v)/F(o)) , apparent quantum (Y), net photosynthetic rate (P(n)), and transpiration rate (T(r)), but increased the light compensation point (LCP) and dark respiration rate (R(d)) of Q. glauca seedlings. T2 inhibited, but T3 played a little enhancement on the aforementioned parameters of Q. glauca seedlings. Under the conditions of medium and light acid rainfall intensities, the above parameters in the three treatments were higher than that of CK, except with lower R(d). The chlorophyll fluorescence and photosynthetic parameters showed a similar tendency in the three treatments, i. e. T2>T3 >T1. It indicated that T1 had the strongest inhibition on seedlings in condition of the severe acid rainfall, while T2 had the most dramatic facilitating effect on seedlings under the medium and light acid rainfall. Intensity of acid rainfall had significant influences on SPAD, F(v)/F(m), F(v)/F(o), Y, P(n), T(r), and maximum photosynthetic rate (A(max)), whereas treatments of acid rainfall affected SPAD, F(v)/F(m), Y, P(n), T(r), A(max) and light saturation point (LSP). The interaction of acid rainfall intensities and treatments played significant effects on SPAD, F(v)/F(m), Y, P(n) and A(max).

  6. Radioecology of Picea excelsa (L.) Lam

    International Nuclear Information System (INIS)

    Sarosiek, J.; Iwen, I.

    1976-01-01

    Within the area of the radiation anomaly near Kowary (Sudeten Mountains) the spruce (Picea excelsa) occurs in habitats with natural soil gamma radiation within 0.05-1.48 mR/h. By way of detailed ecological analysis 5 stentopic habitats of spruce development were selected differing in radiation intensity. In these habitats the intrapopulation variability, radioactivity and ecological properties of the Picea excelsa populations were investigated. It was demonstrated that radiation within the above mentioned range is an essential ecological limiting factor, conditioning ecotypic differences in Picea excelsa. The limiting influence of radiation is manifested in a high frequency of development anomalies in the spruce population. (author)

  7. Radioecology of Picea excelsa (L. Lam

    Directory of Open Access Journals (Sweden)

    Jan Sarosiek

    2015-01-01

    Full Text Available Within the area of the radiation anomaly near Kowary (Sudeten Mountains the spruce (Picea excelsa occurs in habitats with natural soil gamma radiation within 0.05-1.48 mR/h. By way of detailed ecological analysis 5 stenotopic habitats of spruce development were selected differing in radiation intensity. In these habitats the intrapopulation variability, radioactivity and ecological properties of the Picea excelsa populations were investigated. It was demonstrated that radiation within the above mentioned range is an essential ecological limiting factor, conditioning ecotypic differences in Picea excelsa. The limiting influence of radiation is manifested in a high frequency of development anomalies in the spruce population.

  8. Effects of irrigation, fertilization and drought on the occurrence of Lophodermium piceae in Picea abies needles

    Energy Technology Data Exchange (ETDEWEB)

    Lehtijaervi, Asko; Barklund, Pia [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Forest Mycology and Pathology

    1999-08-01

    Effects of irrigation, drought and fertilization on the frequency of the fungal endophyte Lophodermium piceae in green needles was assessed in a 30-year-old experimental stand of Picea abies in southern Sweden. Frequencies of needles with L. piceae were lower in irrigation and ammonium sulphate fertilization treatments than in the control. Drought treatment frequencies were similar to the control. Needles were susceptible to colonization for at least 3 years; colonization increased with needle age. The results indicate that the increased availability of water to the root system as well as ammonium sulphate fertilization indirectly delays colonization of needles by L. piceae 21 refs, 3 figs, 2 tabs

  9. Inheritance of Budbreak and Correlation with Early Height Growth in White Spruce (Picea glauca) from New England

    Science.gov (United States)

    Ronald C. Wilkinson

    1977-01-01

    Variation in budbreak date among 37 half-sib families of white spruce in a replicated one-parent progeny test plantation in southern Maine was only 5 days. Differences in the mean date of budbreak between years were greater than those between families, but the genetic correlation between date of budbreak in different years was .661. Heritability estimates ranged from ....

  10. Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.

    Directory of Open Access Journals (Sweden)

    Dullat Harpreet K

    2011-03-01

    Full Text Available Abstract Background In conifers, terpene synthases (TPSs of the gymnosperm-specific TPS-d subfamily form a diverse array of mono-, sesqui-, and diterpenoid compounds, which are components of the oleoresin secretions and volatile emissions. These compounds contribute to defence against herbivores and pathogens and perhaps also protect against abiotic stress. Results The availability of extensive transcriptome resources in the form of expressed sequence tags (ESTs and full-length cDNAs in several spruce (Picea species allowed us to estimate that a conifer genome contains at least 69 unique and transcriptionally active TPS genes. This number is comparable to the number of TPSs found in any of the sequenced and well-annotated angiosperm genomes. We functionally characterized a total of 21 spruce TPSs: 12 from Sitka spruce (P. sitchensis, 5 from white spruce (P. glauca, and 4 from hybrid white spruce (P. glauca × P. engelmannii, which included 15 monoterpene synthases, 4 sesquiterpene synthases, and 2 diterpene synthases. Conclusions The functional diversity of these characterized TPSs parallels the diversity of terpenoids found in the oleoresin and volatile emissions of Sitka spruce and provides a context for understanding this chemical diversity at the molecular and mechanistic levels. The comparative characterization of Sitka spruce and Norway spruce diterpene synthases revealed the natural occurrence of TPS sequence variants between closely related spruce species, confirming a previous prediction from site-directed mutagenesis and modelling.

  11. Origin and speciation of Picea schrenkiana and Picea smithiana in the Center Asian Highlands and Himalayas

    OpenAIRE

    Li, Lili; Sun, Yongshuai; Zou, Jiabin; Yue, Wei; Wang, Xi; Liu, Jianquan

    2014-01-01

    Elucidating the evolutionary history of current species diversity, especially trees with large effective population sizes and long generation times, is a complicated exercise confounded by gene flow and incomplete lineage sorting. In the present study, we aim to determine the origin and speciation of Picea schrenkiana and Picea smithiana using population genetic data from chloroplast (cp), mitochondrial (mt), and nuclear (nr) genomes. These two species occur in the Central Asian Highlands and...

  12. Transcriptome profiling in conifers and the PiceaGenExpress database show patterns of diversification within gene families and interspecific conservation in vascular gene expression

    Directory of Open Access Journals (Sweden)

    Raherison Elie

    2012-08-01

    Full Text Available Abstract Background Conifers have very large genomes (13 to 30 Gigabases that are mostly uncharacterized although extensive cDNA resources have recently become available. This report presents a global overview of transcriptome variation in a conifer tree and documents conservation and diversity of gene expression patterns among major vegetative tissues. Results An oligonucleotide microarray was developed from Picea glauca and P. sitchensis cDNA datasets. It represents 23,853 unique genes and was shown to be suitable for transcriptome profiling in several species. A comparison of secondary xylem and phelloderm tissues showed that preferential expression in these vascular tissues was highly conserved among Picea spp. RNA-Sequencing strongly confirmed tissue preferential expression and provided a robust validation of the microarray design. A small database of transcription profiles called PiceaGenExpress was developed from over 150 hybridizations spanning eight major tissue types. In total, transcripts were detected for 92% of the genes on the microarray, in at least one tissue. Non-annotated genes were predominantly expressed at low levels in fewer tissues than genes of known or predicted function. Diversity of expression within gene families may be rapidly assessed from PiceaGenExpress. In conifer trees, dehydrins and late embryogenesis abundant (LEA osmotic regulation proteins occur in large gene families compared to angiosperms. Strong contrasts and low diversity was observed in the dehydrin family, while diverse patterns suggested a greater degree of diversification among LEAs. Conclusion Together, the oligonucleotide microarray and the PiceaGenExpress database represent the first resource of this kind for gymnosperm plants. The spruce transcriptome analysis reported here is expected to accelerate genetic studies in the large and important group comprised of conifer trees.

  13. Development of highly reliable in silico SNP resource and genotyping assay from exome capture and sequencing: an example from black spruce (Picea mariana).

    Science.gov (United States)

    Pavy, Nathalie; Gagnon, France; Deschênes, Astrid; Boyle, Brian; Beaulieu, Jean; Bousquet, Jean

    2016-03-01

    Picea mariana is a widely distributed boreal conifer across Canada and the subject of advanced breeding programmes for which population genomics and genomic selection approaches are being developed. Targeted sequencing was achieved after capturing P. mariana exome with probes designed from the sequenced transcriptome of Picea glauca, a distant relative. A high capture efficiency of 75.9% was reached although spruce has a complex and large genome including gene sequences interspersed by some long introns. The results confirmed the relevance of using probes from congeneric species to perform successfully interspecific exome capture in the genus Picea. A bioinformatics pipeline was developed including stringent criteria that helped detect a set of 97,075 highly reliable in silico SNPs. These SNPs were distributed across 14,909 genes. Part of an Infinium iSelect array was used to estimate the rate of true positives by validating 4267 of the predicted in silico SNPs by genotyping trees from P. mariana populations. The true positive rate was 96.2% for in silico SNPs, compared to a genotyping success rate of 96.7% for a set 1115 P. mariana control SNPs recycled from previous genotyping arrays. These results indicate the high success rate of the genotyping array and the relevance of the selection criteria used to delineate the new P. mariana in silico SNP resource. Furthermore, in silico SNPs were generally of medium to high frequency in natural populations, thus providing high informative value for future population genomics applications. © 2015 John Wiley & Sons Ltd.

  14. Symbiotic and nonsymbiotic hemoglobin genes of Casuarina glauca

    DEFF Research Database (Denmark)

    Jacobsen-Lyon, K; Jensen, Erik Østergaard; Jørgensen, Jan-Elo

    1995-01-01

    Casuarina glauca has a gene encoding hemoglobin (cashb-nonsym). This gene is expressed in a number of plant tissues. Casuarina also has a second family of hemoglobin genes (cashb-sym) expressed at a high level in the nodules that Casuarina forms in a nitrogen-fixing symbiosis with the actinomycete...... of the Casuarina gene. The finding that the nonsymbiotic Casuarina gene is also correctly expressed in L. corniculatus suggests to us that a comparable non-symbiotic hemoglobin gene will be found in legume species. Udgivelsesdato: 1995-Feb...

  15. Composition and Biological Activity of Picea pungens and Picea orientalis Seed and Cone Essential Oils.

    Science.gov (United States)

    Wajs-Bonikowska, Anna; Szoka, Łukasz; Karna, Ewa; Wiktorowska-Owczarek, Anna; Sienkiewicz, Monika

    2017-03-01

    The increasing consumption of natural products lead us to discover and study new plant materials, such as conifer seeds and cones, which could be easily available from the forest industry as a waste material, for their potential uses. The chemical composition of the essential oils of Picea pungens and Picea orientalis was fully characterized by GC and GC/MS methods. Seed and cone oils of both tree species were composed mainly of monoterpene hydrocarbons, among which limonene, α- and β-pinene were the major, but in different proportions in the examined conifer essential oils. The levorotary form of chiral monoterpene molecules was predominant over the dextrorotary form. The composition of oils from P. pungens seeds and cones was similar, while the hydrodistilled oils of P. orientalis seeds and cones differed from each other, mainly by a higher amount of oxygenated derivatives of monoterpenes and by other higher molar mass terpenes in seed oil. The essential oils showed mild antimicrobial action, however P. orientalis cone oil exhibited stronger antimicrobial properties against tested bacterial species than those of P. pungens. Effects of the tested cone essential oils on human skin fibroblasts and microvascular endothelial cells (HMEC-1) were similar: in a concentration of 0 - 0.075 μl/ml the oils were rather safe for human skin fibroblasts and 0 - 0.005 μl/ml for HMEC-1 cells. IC 50 value of Picea pungens oils was 0.115 μl/ml, while that of Picea orientalis was 0.105 μl/ml. The value of IC 50 of both oils were 0.035 μl/ml for HMEC-1 cells. The strongest effect on cell viability had the oil from Picea orientalis cones, while on DNA synthesis the oil from Picea pungens cones. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  16. End Sequencing and Finger Printing of Human & Mouse BAC Libraries

    Energy Technology Data Exchange (ETDEWEB)

    Fraser, C

    2005-09-27

    This project provided for continued end sequencing of existing and new BAC libraries constructed to support human sequencing as well as to initiate BAC end sequencing from the mouse BAC libraries constructed to support mouse sequencing. The clones, the sequences, and the fingerprints are now an available resource for the community at large. Research and development of new metaodologies for BAC end sequencing have reduced costs and increase throughput.

  17. A set of BAC clones spanning the human genome.

    NARCIS (Netherlands)

    Krzywinski, M.; Bosdet, I.; Smailus, D.; Chiu, R.; Mathewson, C.; Wye, N.; Barber, S.; Brown-John, M.; Chan, S.; Chand, S.; Cloutier, A.; Girn, N.; Lee, D.; Masson, A.; Mayo, M.; Olson, T.; Pandoh, P.; Prabhu, A.L.; Schoenmakers, E.F.P.M.; Tsai, M.Y.; Albertson, D.; Lam, W.W.; Choy, C.O.; Osoegawa, K.; Zhao, S.; Jong, P.J. de; Schein, J.; Jones, S.; Marra, M.A.

    2004-01-01

    Using the human bacterial artificial chromosome (BAC) fingerprint-based physical map, genome sequence assembly and BAC end sequences, we have generated a fingerprint-validated set of 32 855 BAC clones spanning the human genome. The clone set provides coverage for at least 98% of the human

  18. Transposon-mediated BAC transgenesis in zebrafish and mice

    Directory of Open Access Journals (Sweden)

    Sumiyama Kenta

    2009-10-01

    Full Text Available Abstract Background Bacterial artificial chromosomes (BACs are among the most widely used tools for studies of gene regulation and function in model vertebrates, yet methods for predictable delivery of BAC transgenes to the genome are currently limited. This is because BAC transgenes are usually microinjected as naked DNA into fertilized eggs and are known to integrate as multi-copy concatamers in the genome. Although conventional methods for BAC transgenesis have been very fruitful, complementary methods for generating single copy BAC integrations would be desirable for many applications. Results We took advantage of the precise cut-and-paste behavior of a natural transposon, Tol2, to develop a new method for BAC transgenesis. In this new method, the minimal sequences of the Tol2 transposon were used to deliver precisely single copies of a ~70 kb BAC transgene to the zebrafish and mouse genomes. We mapped the BAC insertion sites in the genome by standard PCR methods and confirmed transposase-mediated integrations. Conclusion The Tol2 transposon has a surprisingly large cargo capacity that can be harnessed for BAC transgenesis. The precise delivery of single-copy BAC transgenes by Tol2 represents a useful complement to conventional BAC transgenesis, and could aid greatly in the production of transgenic fish and mice for genomics projects, especially those in which single-copy integrations are desired.

  19. Phytochemical Analysis and Free Radical Scavenging Activity of Medicinal Plants Gnidia glauca and Dioscorea bulbifera

    Science.gov (United States)

    Ghosh, Sougata; Derle, Abhishek; Ahire, Mehul; More, Piyush; Jagtap, Soham; Phadatare, Suvarna D.; Patil, Ajay B.; Jabgunde, Amit M.; Sharma, Geeta K.; Shinde, Vaishali S.; Pardesi, Karishma; Dhavale, Dilip D.; Chopade, Balu A.

    2013-01-01

    Gnidia glauca and Dioscorea bulbifera are traditional medicinal plants that can be considered as sources of natural antioxidants. Herein we report the phytochemical analysis and free radical scavenging activity of their sequential extracts. Phenolic and flavonoid content were determined. Scavenging activity was checked against pulse radiolysis generated ABTS•+ and OH radical, in addition to DPPH, superoxide and hydroxyl radicals by biochemical methods followed by principal component analysis. G. glauca leaf extracts were rich in phenolic and flavonoid content. Ethyl acetate extract of D. bulbifera bulbs and methanol extract of G. glauca stem exhibited excellent scavenging of pulse radiolysis generated ABTS•+ radical with a second order rate constant of 2.33×106 and 1.72×106, respectively. Similarly, methanol extract of G. glauca flower and ethyl acetate extract of D. bulbifera bulb with second order rate constants of 4.48×106 and 4.46×106 were found to be potent scavengers of pulse radiolysis generated OH radical. G. glauca leaf and stem showed excellent reducing activity and free radical scavenging activity. HPTLC fingerprinting, carried out in mobile phase, chloroform: toluene: ethanol (4: 4: 1, v/v) showed presence of florescent compound at 366 nm as well as UV active compound at 254 nm. GC-TOF-MS analysis revealed the predominance of diphenyl sulfone as major compound in G. glauca. Significant levels of n-hexadecanoic acid and octadecanoic acid were also present. Diosgenin (C27H42O3) and diosgenin (3á,25R) acetate were present as major phytoconstituents in the extracts of D. bulbifera. G. glauca and D. bulbifera contain significant amounts of phytochemicals with antioxidative properties that can be exploited as a potential source for herbal remedy for oxidative stress induced diseases. These results rationalize further investigation in the potential discovery of new natural bioactive principles from these two important medicinal plants. PMID:24367520

  20. [Phenolic acid derivatives from Bauhinia glauca subsp. pernervosa].

    Science.gov (United States)

    Zhao, Qiao-Li; Wu, Zeng-Bao; Zheng, Zhi-Hui; Lu, Xin-Hua; Liang, Hong; Cheng, Wei; Zhang, Qing-Ying; Zhao, Yu-Ying

    2011-08-01

    To study the chemical constituents of Bauhinia glauca subsp. pernervosa, eleven phenolic acids were isolated from a 95% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, MCI, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS, these compounds were identified as isopropyl O-beta-(6'-O-galloyl)-glucopyranoside (1), ethyl O-beta-(6'-O-galloyl)-glucopyranoside (2), 3, 4, 5-trimethoxyphenyl-(6'-O-galloyl)-O-beta-D-glucopyranoside (3), 3, 4, 5-trimethoxyphenyl-beta-D-glucopyranoside (4), gallic acid (5), methyl gallate (6), ethyl gallate (7), protocatechuic acid (8), 3, 5-dimethoxy-4-hydroxybenzoic acid (9), erigeside C (10) and glucosyringic acid (11). Among them, compound 1 is a new polyhydroxyl compound; compounds 2, 10, and 11 were isolated from the genus Bauhinia for the first time, and the other compounds were isolated from the plant for the first time. Compounds 6 and 8 showed significant protein tyrosine phosphatase1B (PTP1B) inhibitory activity in vitro with the IC50 values of 72.3 and 54.1 micromol x L(-1), respectively.

  1. MORFOGÉNESIS IN VITRO DE Pseudotsuga menziesii VAR. glauca

    Directory of Open Access Journals (Sweden)

    María Guadalupe Carrillo-Benítez

    2011-01-01

    Full Text Available Se evaluó la respuesta morfogénica a partir de embriones cigóticos cultivados in vitro de semilla almacenada (un año de Pseudotsuga menziesii var. glauca recolectada en Tlaxcala. Las semillas fueron desinfestadas con detergente y H2O2 (3 % v/v durante 48 h en agitación a 50 rpm, cultivadas en el medio de Murashige y Skoog (1962 sin reguladores. La germinación ocurrió después de siete días y posteriormente subcultivados a un medio MS con 2,4-D (3 mg·L-1 y BA (1 mg·L-1. Con los callos obtenidos en un medio HS se evaluaron tres concentraciones de ABA para promover formación de estructuras embrionarias, presentándose el mejor tratamiento con concentración de 10.0 mg·L-1 (P<.0001. El mejor desarrollo de plántulas se presentó empleando un medio Murashige y Skoog (1962 con sacarosa al 6 %. Se usaron micorrizas para mejor adaptación de plántulas a suelo. No hubo formación de raíces.

  2. Generation of BAC transgenic epithelial organoids.

    Directory of Open Access Journals (Sweden)

    Gerald Schwank

    Full Text Available Under previously developed culture conditions, mouse and human intestinal epithelia can be cultured and expanded over long periods. These so-called organoids recapitulate the three-dimensional architecture of the gut epithelium, and consist of all major intestinal cell types. One key advantage of these ex vivo cultures is their accessibility to live imaging. So far the establishment of transgenic fluorescent reporter organoids has required the generation of transgenic mice, a laborious and time-consuming process, which cannot be extended to human cultures. Here we present a transfection protocol that enables the generation of recombinant mouse and human reporter organoids using BAC (bacterial artificial chromosome technology.

  3. Movements of Blue Sharks (Prionace glauca) across Their Life History

    Science.gov (United States)

    Vandeperre, Frederic; Aires-da-Silva, Alexandre; Fontes, Jorge; Santos, Marco; Serrão Santos, Ricardo; Afonso, Pedro

    2014-01-01

    Spatial structuring and segregation by sex and size is considered to be an intrinsic attribute of shark populations. These spatial patterns remain poorly understood, particularly for oceanic species such as blue shark (Prionace glauca), despite its importance for the management and conservation of this highly migratory species. This study presents the results of a long-term electronic tagging experiment to investigate the migratory patterns of blue shark, to elucidate how these patterns change across its life history and to assess the existence of a nursery area in the central North Atlantic. Blue sharks belonging to different life stages (n = 34) were tracked for periods up to 952 days during which they moved extensively (up to an estimated 28.139 km), occupying large parts of the oceanic basin. Notwithstanding a large individual variability, there were pronounced differences in movements and space use across the species' life history. The study provides strong evidence for the existence of a discrete central North Atlantic nursery, where juveniles can reside for up to at least 2 years. In contrast with previously described nurseries of coastal and semi-pelagic sharks, this oceanic nursery is comparatively vast and open suggesting that shelter from predators is not its main function. Subsequently, male and female blue sharks spatially segregate. Females engage in seasonal latitudinal migrations until approaching maturity, when they undergo an ontogenic habitat shift towards tropical latitudes. In contrast, juvenile males generally expanded their range southward and apparently displayed a higher degree of behavioural polymorphism. These results provide important insights into the spatial ecology of pelagic sharks, with implications for the sustainable management of this heavily exploited shark, especially in the central North Atlantic where the presence of a nursery and the seasonal overlap and alternation of different life stages coincides with a high fishing

  4. Genetic stability of pestivirus genomes cloned into BACs

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Reimann, Ilona; Uttenthal, Åse

    pestivirus genomes to demonstrate the suitability of the BAC vector for harbouring pestivirus genomes. Two BAC clones, comprising the complete genomes of BDV Gifhorn (pBeloGif3) and CSFV Paderborn (pBeloPader10) were passaged 15 times in E.coli representing at least 360 bacteria generations. From 15th...

  5. Typha angustifolia L. X T. latifolia L. (T.) (glauca Godr.) in Nederland

    NARCIS (Netherlands)

    Ooststroom, van S.J.; Reichgelt, Th.J.

    1962-01-01

    Typha x glauca Godr., the hybrid of T. angustifolia L. and T. latifolia L. was found in the Netherlands for the first time in 1847 and again in 1946, as appears from the collections in the Rijksherbarium, Leiden. Fig. 1 shows the differences found in the female flowers, the pollen, and the

  6. Detecting Sulfuric and Nitric Acid Rain Stresses on Quercus glauca through Hyperspectral Responses.

    Science.gov (United States)

    Wang, Shanqian; Zhang, Xiuying; Ma, Yuandan; Li, Xinhui; Cheng, Min; Zhang, Xiaomin; Liu, Lei

    2018-03-09

    Acid rain, which has become one of the most severe global environmental issues, is detrimental to plant growth. However, effective methods for monitoring plant responses to acid rain stress are currently lacking. The hyperspectral technique provides a cost-effective and nondestructive way to diagnose acid rain stresses. Taking a widely distributed species ( Quercus glauca ) in Southern China as an example, this study aims to monitor the hyperspectral responses of Q. glauca to simulated sulfuric acid rain (SAR) and nitric acid rain (NAR). A total of 15 periods of leaf hyperspectral data under four pH levels of SAR and NAR were obtained during the experiment. The results showed that hyperspectral information could be used to distinguish plant responses under acid rain stress. An index (green peak area index, GPAI) was proposed to indicate acid rain stresses, based on the significantly variations in the region of 500-660 nm. Light acid rain (pH 4.5 SAR and NAR) promoted Q. glauca growth relative to the control groups (pH 5.6 SAR and NAR); moderate acid rain (pH 3.0 SAR) firstly promoted and then inhibited plant growth, while pH 3.0 NAR showed mild inhibitory effects during the experiment; and heavy acid rain (pH 2.0) significantly inhibited plant growth. Compared with NAR, SAR induced more serious damages to Q. glauca . These results could help monitor acid rain stress on plants on a regional scale using remote sensing techniques.

  7. Pollinator networks, alien species and the conservation of rare plants: Trinia glauca as a case study

    NARCIS (Netherlands)

    Carvalheiro, L.G.; Barbosa, E.R.; Memmott, J.

    2008-01-01

    1. Despite the essential role of pollination in the maintenance of many rare plant species, conservation management plans rarely consider the service of pollination. 2. This study identifies the main pollinators of a rare English plant species, Trinia glauca (Apiaceae), and provides recommendations

  8. Construction of BAC Libraries from Flow-Sorted Chromosomes.

    Science.gov (United States)

    Šafář, Jan; Šimková, Hana; Doležel, Jaroslav

    2016-01-01

    Cloned DNA libraries in bacterial artificial chromosome (BAC) are the most widely used form of large-insert DNA libraries. BAC libraries are typically represented by ordered clones derived from genomic DNA of a particular organism. In the case of large eukaryotic genomes, whole-genome libraries consist of a hundred thousand to a million clones, which make their handling and screening a daunting task. The labor and cost of working with whole-genome libraries can be greatly reduced by constructing a library derived from a smaller part of the genome. Here we describe construction of BAC libraries from mitotic chromosomes purified by flow cytometric sorting. Chromosome-specific BAC libraries facilitate positional gene cloning, physical mapping, and sequencing in complex plant genomes.

  9. The need for blood alcohol concentration (BAC) legislation in Nigeria

    African Journals Online (AJOL)

    Dr Patrick O Erah

    Tropical Journal of Pharmaceutical Research, June 2004; 3 (1): 319-327. © Pharmacotherapy Group, ... 1Department of Restorative Dentistry, Clinical therapeutic Unit, University of Benin, Benin City, .... An international survey of BAC is quite.

  10. Allozyme variation in Picea mariana from Newfoundland: Discussion

    Science.gov (United States)

    William B. Critchfield

    1987-01-01

    In their recent paper describing the distribution of genetic variation in black spruce (Picea mariana (Mill.) B.S.P.) on the island of Newfoundland, Yeh et el. (1986) concluded that a center of variability in west-central Newfoundland derived from ancestral populations that persisted on the island during the last (Wisconsin) glaciation. They...

  11. How wind affects growth in treeline Picea abies

    Czech Academy of Sciences Publication Activity Database

    Kašpar, J.; Hošek, Jiří; Treml, V.

    2017-01-01

    Roč. 127, č. 2 (2017), s. 1-12 ISSN 1664-2201 Institutional support: RVO:68378289 Keywords : height increment * picea abies * radial growth * thigmomorphogenesis * tree rings Subject RIV: GK - Forestry OBOR OECD: Forestry Impact factor: 2.281, year: 2016 https://link.springer.com/content/pdf/10.1007%2Fs00035-017-0186-x.pdf

  12. Ethanol exposure can inhibit red spruce ( Picea rubens ) seed germination

    Science.gov (United States)

    John R. Butnor; Brittany M. Verrico; Victor Vankus; Stephen R. Keller

    2018-01-01

    Flotation of seeds in solvents is a common means of separating unfilled and filled seeds. While a few protocols for processing red spruce (Picea rubens) seeds recommend ethanol flotation, delayed and reduced germination have been reported. We conducted an ethanol bioassay on seeds previously stored at -20°C to quantify the concentration required to separate red spruce...

  13. Variation in phenology and monoterpene patterns of defoliated and nondefoliated Douglas-fir (Pseudotsuga menziesii var. glauca).

    Science.gov (United States)

    Rose-Marie Muzika; Judith Engle; Catherine Parks; Boyd. Wickman

    1993-01-01

    Foliage was collected from paired Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) trees characterized as either "resistant" or "susceptible" western spruce budworm (Choristoneura occidentalis Freeman (Lepidoptera: Tortricidae) attack. Resistant trees produced more...

  14. Transposon-mediated BAC transgenesis in human ES cells

    Science.gov (United States)

    Rostovskaya, Maria; Fu, Jun; Obst, Mandy; Baer, Isabell; Weidlich, Stefanie; Wang, Hailong; Smith, Andrew J. H.; Anastassiadis, Konstantinos; Stewart, A. Francis

    2012-01-01

    Transgenesis is a cornerstone of molecular biology. The ability to integrate a specifically engineered piece of DNA into the genome of a living system is fundamental to our efforts to understand life and exploit its implications for medicine, nanotechnology and bioprospecting. However, transgenesis has been hampered by position effects and multi-copy integration problems, which are mainly due to the use of small, plasmid-based transgenes. Large transgenes based on native genomic regions cloned into bacterial artificial chromosomes (BACs) circumvent these problems but are prone to fragmentation. Herein, we report that contrary to widely held notions, large BAC-sized constructs do not prohibit transposition. We also report the first reliable method for BAC transgenesis in human embryonic stem cells (hESCs). The PiggyBac or Sleeping Beauty transposon inverted repeats were integrated into BAC vectors by recombineering, followed by co-lipofection with the corresponding transposase in hESCs to generate robust fluorescent protein reporter lines for OCT4, NANOG, GATA4 and PAX6. BAC transposition delivers several advantages, including increased frequencies of single-copy, full-length integration, which will be useful in all transgenic systems but especially in difficult venues like hESCs. PMID:22753106

  15. Efficient Expression of Acetylcholine-Binding Protein from Aplysia californica in Bac-to-Bac System

    Directory of Open Access Journals (Sweden)

    Bo Lin

    2014-01-01

    Full Text Available The Bac-to-Bac baculovirus expression system can efficiently produce recombinant proteins, but the system may have to be optimized to achieve high-level expression for different candidate proteins. We reported here the efficient expression of acetylcholine-binding proteins from sea hares Aplysia californica (Ac-AChBP and a convenient method to monitor protein expression level in this expression system. Three key factors affecting expression of Ac-AChBP were optimized for maximizing the yield, which included the cell density, volume of the infecting baculovirus inoculums, and the culturing time of postinfection. We have found it to reach a high yield of ∼5 mg/L, which needs 55 h incubation after infection at the cell density of 2 × 106 cells/mL with an inoculum volume ratio of 1 : 100. The optimized expression system in this study was also applied for expressing another protein Ls-AChBP from Lymnaea stagnalis successfully. Therefore, this established method is helpful to produce high yields of AChBP proteins for X-ray crystallographic structural and functional studies.

  16. An efficient approach to BAC based assembly of complex genomes.

    Science.gov (United States)

    Visendi, Paul; Berkman, Paul J; Hayashi, Satomi; Golicz, Agnieszka A; Bayer, Philipp E; Ruperao, Pradeep; Hurgobin, Bhavna; Montenegro, Juan; Chan, Chon-Kit Kenneth; Staňková, Helena; Batley, Jacqueline; Šimková, Hana; Doležel, Jaroslav; Edwards, David

    2016-01-01

    There has been an exponential growth in the number of genome sequencing projects since the introduction of next generation DNA sequencing technologies. Genome projects have increasingly involved assembly of whole genome data which produces inferior assemblies compared to traditional Sanger sequencing of genomic fragments cloned into bacterial artificial chromosomes (BACs). While whole genome shotgun sequencing using next generation sequencing (NGS) is relatively fast and inexpensive, this method is extremely challenging for highly complex genomes, where polyploidy or high repeat content confounds accurate assembly, or where a highly accurate 'gold' reference is required. Several attempts have been made to improve genome sequencing approaches by incorporating NGS methods, to variable success. We present the application of a novel BAC sequencing approach which combines indexed pools of BACs, Illumina paired read sequencing, a sequence assembler specifically designed for complex BAC assembly, and a custom bioinformatics pipeline. We demonstrate this method by sequencing and assembling BAC cloned fragments from bread wheat and sugarcane genomes. We demonstrate that our assembly approach is accurate, robust, cost effective and scalable, with applications for complete genome sequencing in large and complex genomes.

  17. Adaptive Evolution and Demographic History of Norway Spruce (Picea Abies)

    OpenAIRE

    Källman, Thomas

    2009-01-01

    One of the major challenges in evolutionary biology is to determine the genetic basis of adaptive variation. In Norway spruce (Picea abies) the timing of bud set shows a very strong latitudinal cline despite a very low genetic differentiation between populations. The timing of bud set in Norway spruce is under strong genetic control and triggered by changes in photoperiod, but no genes controlling this response have so far been described. In this thesis we used a combination of functional stu...

  18. Sequencing of the needle transcriptome from Norway spruce (Picea abies Karst L. reveals lower substitution rates, but similar selective constraints in gymnosperms and angiosperms

    Directory of Open Access Journals (Sweden)

    Chen Jun

    2012-11-01

    Full Text Available Abstract Background A detailed knowledge about spatial and temporal gene expression is important for understanding both the function of genes and their evolution. For the vast majority of species, transcriptomes are still largely uncharacterized and even in those where substantial information is available it is often in the form of partially sequenced transcriptomes. With the development of next generation sequencing, a single experiment can now simultaneously identify the transcribed part of a species genome and estimate levels of gene expression. Results mRNA from actively growing needles of Norway spruce (Picea abies was sequenced using next generation sequencing technology. In total, close to 70 million fragments with a length of 76 bp were sequenced resulting in 5 Gbp of raw data. A de novo assembly of these reads, together with publicly available expressed sequence tag (EST data from Norway spruce, was used to create a reference transcriptome. Of the 38,419 PUTs (putative unique transcripts longer than 150 bp in this reference assembly, 83.5% show similarity to ESTs from other spruce species and of the remaining PUTs, 3,704 show similarity to protein sequences from other plant species, leaving 4,167 PUTs with limited similarity to currently available plant proteins. By predicting coding frames and comparing not only the Norway spruce PUTs, but also PUTs from the close relatives Picea glauca and Picea sitchensis to both Pinus taeda and Taxus mairei, we obtained estimates of synonymous and non-synonymous divergence among conifer species. In addition, we detected close to 15,000 SNPs of high quality and estimated gene expression differences between samples collected under dark and light conditions. Conclusions Our study yielded a large number of single nucleotide polymorphisms as well as estimates of gene expression on transcriptome scale. In agreement with a recent study we find that the synonymous substitution rate per year (0.6 × 10

  19. [Effect of ectomycorrhizae on the growth of Picea koraiensis seedlings].

    Science.gov (United States)

    Song, Rui-Qing; Wu, Ke

    2005-12-01

    Basidioscarps of Agaricales in different Picea koraiensis forest plantations were collected during August-October, 2000. 36 isolaters of species of Agaricales were obtained by isolating and culturing to the basidioscarps. Through indoor inoculation test on seedlings of Picea koraiensis, 6 ectomycorrhizae fungi cultures were obtained from 36 isolaters. The inoculation results show that the period for ectomycorrhizae inoculation to 1-year seedlings of Picea koraiensis should be about 30 days after seedlings emerging, the suitable temperature for ectomycorrhizae forming is about 20 degrees C. 6 ectomycorrhizae strains all have growth-promoting effect to the seedlings of Picea koraiensis. The contents of chlorophyll a of the seedlings inoculated strains of Agaricus silvaticus, 031 and L15 were significantly higher than other strains and control. The contents of chlorophyll b in the seedlings inoculated strains 009, 004, Agaricus silvaticus and L15 were significantly higher than other strains and control. The weights of seedlings which inoculated strains 009, 025, 031, Agaricus silvaticus and L15 were significantly different to control, the weight of seedlings inoculated strains of Agaricus silvaticus and L15 are 19.23% and 23.08% more than control; The heights of the seedlings inoculated 6 strains all have significant difference to control, the weight of seedlings inoculated strains of Agaricus silvaticus and L15 are 17.83% and 16.37% more than control. The results of outdoor inoculation show that the seedlings inoculated Agaricus silvaticus grow best on height, 9.25% more than control after inoculated 70 days; the seedlings inoculated strain L15 grow best on collar diameter, 9.92% more than control after inoculated 70 days; the lateral root numbers of seedlings inoculated strain 009 is largest, 51.91% more than control after inoculated 70 days; the main roots of seedlings inoculated strain 009 are longest, 3.36% more than control after inoculated 70 days; the

  20. The need for blood alcohol concentration (BAC) legislation in Nigeria

    African Journals Online (AJOL)

    The pharmacology, clinical and sports implications of indulgence in alcohol and the debate on its legal status are highlighted in this article. The information presented could offer both clinical and safety benefits to psychomotor tasks executors and road safety professionals. Keywords: Blood alcohol concentration (BAC), ...

  1. An efficient approach to BAC based assembly of complex genomes

    Czech Academy of Sciences Publication Activity Database

    Visendi, P.; Berkman, P.J.; Hayashi, S.; Golicz, A.A.; Bayer, P.E.; Ruperao, P.; Hurgobin, B.; Montenegro, J.; Chan, C.K.K.; Staňková, Helena; Batley, J.; Šimková, Hana; Doležel, Jaroslav; Edwards, D.

    2016-01-01

    Roč. 12, JAN 20 (2016), s. 2 ISSN 1746-4811 R&D Projects: GA MŠk(CZ) LO1204; GA ČR(CZ) GAP501/12/2554 Institutional support: RVO:61389030 Keywords : Next-generation sequencing * SASSY * BAC Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.510, year: 2016

  2. Phytochemical composition and antinociceptive activity of Bauhinia glauca subsp. hupehana in rats.

    Science.gov (United States)

    Xu, Jinlong; Zhao, Qizhi; Wei, Lei; Yang, Yu; Xu, Rui; Yu, Nengjiang; Zhao, Yimin

    2015-01-01

    In traditional medicine, Bauhinia glauca subsp. hupehana has long been used as an analgesic agent in China. The aim of this study was to evaluate the antinociceptive activity of the ethanol extract of the aerial parts of B. glauca subsp. hupehana (BHE) in rats and its chemical fingerprint. The antinociceptive activity of BHE was assessed in mice using chemically and heat-induced pain models, such as the acetic acid-induced writhing, hot plate, tail-flick and glutamate tests. Naltrexone hydrochloride, a non-selective opioid receptor antagonist, was utilized to determine the involvement of the opioid system. In addition to this, the involvements of the cGMP and ATP-sensitive K+ channel pathways were also detected using methylene blue and glibenclamide. The oral administration of BHE (at doses of 50, 100 and 200 mg/kg) produced significant and dose-related inhibitions in both the chemically and heat-induced pain models. Interestingly, in the abdominal constriction test, when the dose of BHE was increased to 800 mg/kg (p.o., n = 10), the inhibition rate was 100%. The antinociceptive mechanism may involve the cGMP pathway and ATP sensitive K+ channel pathway. The central antinociceptive effect was not antagonized by naltrexone. One phenolic acid, one lignin and five flavonoids were isolated from BHE. The antinociceptive activity of BHE was most likely due to the presence of the flavonoids. The acute toxicity results showed that BHE was safe at a high dose (2 g/kg, p.o.). The current investigation demonstrates that B. glauca subsp. hupehana is a potential candidate for the development of novel, non-opioid, analgesic phytomedicines.

  3. Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants.

    Science.gov (United States)

    Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

    2012-07-01

    The objectives of the present work were in vitro propagation of Araucaria excelsa R. Br. var. glauca Carrière (Norfolk Island pine) with focus on the evaluation of the mean number of shoots per explant (MNS/E) and mean length of shoots per explants (MLS/E) produced by different parts of the orthotropic stem of A. excelsa R. Br. var. glauca in response to plant growth regulators. Norfolk Island pine axillary meristems responded very well to the 2-iso-pentenyl adenine (2iP) and thidiazuron (TDZ) levels. Explants taken from stem upper segments in the media containing 2iP had a higher MNS/E (3.47) and MLS/E (6.27 mm) in comparison to those taken from stem lower segments, which were 0.71 and 0.51 mm, respectively. Using 0.045 μM TDZ in the MS medium not only resulted in 4.60 MNS/E with 7.08 mm MLS/E but proliferated shoots showed a good performance as well. Investigating the best position of stem explant on mother plant as well as the best concentrations of growth regulators were performed which were useful for efficient micropropagation of this plant. Thirty three percent of explants were rooted in the MS medium containing 3 % sucrose, supplemented with 7.5 μM of both NAA and IBA for 2 weeks before transferring to a half strength MS medium without any growth regulator. Plantlets obtained were acclimatized and transferred to the greenhouse with less than 20 % mortality. This procedure considered the first successful report for regeneration and acclimatization of A. excelsa R. Br. var. glauca plantlet through main stem explants.

  4. [Effects of simulated acid rain on Quercus glauca seedlings photosynthesis and chlorophyll fluorescence].

    Science.gov (United States)

    Li, Jia; Jiang, Hong; Yu, Shu-quan; Jiang, Fu-wei; Yin, Xiu-min; Lu, Mei-juan

    2009-09-01

    Taking the seedlings of Quercus glauca, a dominant evergreen broadleaf tree species in subtropical area, as test materials, this paper studied their photosynthesis, chlorophyll fluorescence, and chlorophyll content under effects of simulated acid rain with pH 2.5, 4.0, and 5.6 (CK). After 2-year acid rain stress, the net photosynthetic rate of Q. glauca increased significantly with decreasing pH of acid rain. The acid rain with pH 2.5 and 4.0 increased the stomatal conductance and transpiration rate, and the effect was more significant under pH 2.5. The intercellular CO2 concentration decreased in the order of pH 2.5 > pH 5.6 > pH 4.0. The maximum photosynthetic rate, light compensation point, light saturation point, and dark respiration rate were significantly higher under pH 2.5 and 4.0 than under pH 5.6, while the apparent quantum yield was not sensitive to acid rain stress. The maximal photochemical efficiency of PS II and the potential activity of PS II under pH 2.5 and 4.0 were significantly higher than those under pH 5.6. The relative chlorophyll content was in the order of pH 2.5 > pH 5.6 > pH 4.0, and there was a significant difference between pH 2.5 and 4.0. All the results suggested that the photosynthesis and chlorophyll fluorescence of Q. glauca increased under the effects of acid rain with pH 2.5 and 4.0, and the acid rain with pH 2.5 had more obvious effects.

  5. Creation of BAC genomic resources for cocoa ( Theobroma cacao L.) for physical mapping of RGA containing BAC clones.

    Science.gov (United States)

    Clément, D; Lanaud, C; Sabau, X; Fouet, O; Le Cunff, L; Ruiz, E; Risterucci, A M; Glaszmann, J C; Piffanelli, P

    2004-05-01

    We have constructed and validated the first cocoa ( Theobroma cacao L.) BAC library, with the aim of developing molecular resources to study the structure and evolution of the genome of this perennial crop. This library contains 36,864 clones with an average insert size of 120 kb, representing approximately ten haploid genome equivalents. It was constructed from the genotype Scavina-6 (Sca-6), a Forastero clone highly resistant to cocoa pathogens and a parent of existing mapping populations. Validation of the BAC library was carried out with a set of 13 genetically-anchored single copy and one duplicated markers. An average of nine BAC clones per probe was identified, giving an initial experimental estimation of the genome coverage represented in the library. Screening of the library with a set of resistance gene analogues (RGAs), previously mapped in cocoa and co-localizing with QTL for resistance to Phytophthora traits, confirmed at the physical level the tight clustering of RGAs in the cocoa genome and provided the first insights into the relationships between genetic and physical distances in the cocoa genome. This library represents an available BAC resource for structural genomic studies or map-based cloning of genes corresponding to important QTLs for agronomic traits such as resistance genes to major cocoa pathogens like Phytophthora spp ( palmivora and megakarya), Crinipellis perniciosa and Moniliophthora roreri.

  6. Rapid construction of a Bacterial Artificial Chromosomal (BAC) expression vector using designer DNA fragments.

    Science.gov (United States)

    Chen, Chao; Zhao, Xinqing; Jin, Yingyu; Zhao, Zongbao Kent; Suh, Joo-Won

    2014-11-01

    Bacterial artificial chromosomal (BAC) vectors are increasingly being used in cloning large DNA fragments containing complex biosynthetic pathways to facilitate heterologous production of microbial metabolites for drug development. To express inserted genes using Streptomyces species as the production hosts, an integration expression cassette is required to be inserted into the BAC vector, which includes genetic elements encoding a phage-specific attachment site, an integrase, an origin of transfer, a selection marker and a promoter. Due to the large sizes of DNA inserted into the BAC vectors, it is normally inefficient and time-consuming to assemble these fragments by routine PCR amplifications and restriction-ligations. Here we present a rapid method to insert fragments to construct BAC-based expression vectors. A DNA fragment of about 130 bp was designed, which contains upstream and downstream homologous sequences of both BAC vector and pIB139 plasmid carrying the whole integration expression cassette. In-Fusion cloning was performed using the designer DNA fragment to modify pIB139, followed by λ-RED-mediated recombination to obtain the BAC-based expression vector. We demonstrated the effectiveness of this method by rapid construction of a BAC-based expression vector with an insert of about 120 kb that contains the entire gene cluster for biosynthesis of immunosuppressant FK506. The empty BAC-based expression vector constructed in this study can be conveniently used for construction of BAC libraries using either microbial pure culture or environmental DNA, and the selected BAC clones can be directly used for heterologous expression. Alternatively, if a BAC library has already been constructed using a commercial BAC vector, the selected BAC vectors can be manipulated using the method described here to get the BAC-based expression vectors with desired gene clusters for heterologous expression. The rapid construction of a BAC-based expression vector facilitates

  7. Genetic diversity and conservation of Picea chihuahuana Martínez ...

    African Journals Online (AJOL)

    SAM

    2014-07-09

    Jul 9, 2014 ... individuals within a population and among populations within a species. Hence ... risks related to loss of species, populations and genetic resources; c) ... this spruce and two other congeners, Picea mexicana. Martínez and Picea .... high levels of self-fertilization and mating between closely related ...

  8. The effect of intermittent dosing of Nicotiana glauca on teratogenesis in goats.

    Science.gov (United States)

    Welch, K D; Panter, K E; Lee, S T; Gardner, D R

    2015-01-01

    Sustained inhibition of fetal movement in livestock species, induced by several poisonous plants, can result in numerous skeletal-contracture malformations. Lupines are responsible for a condition in cattle referred to as "crooked calf syndrome" that occurs when pregnant cattle graze teratogenic lupines. Similar malformations are also seen in animals poisoned by Conium maculatum (coniine) and Nicotiana glauca (anabasine). A proposed management strategy to limit these types of birth defects includes utilizing an intermittent grazing schedule to allow short durations of grazing lupine-infested areas interrupted by movement to a lupine-free pasture. The objective of this study was to use a goat model to determine if an intermittent schedule of five continuous days on treatment followed by two days off treatment would be sufficient to decrease, or prevent, the incidence of anabasine-induced malformations. The data from this study suggest that, for N. glauca in goats, the intermittent grazing program of five days exposure with two days of non-exposure is insufficient to prevent significant skeletal malformations from occurring. However, this study did demonstrate an inverse relationship between the amount of serum anabasine in the dam and the extent of fetal movement. Published by Elsevier Ltd.

  9. Interspecific Plastome Recombination Reflects Ancient Reticulate Evolution in Picea (Pinaceae).

    Science.gov (United States)

    Sullivan, Alexis R; Schiffthaler, Bastian; Thompson, Stacey Lee; Street, Nathaniel R; Wang, Xiao-Ru

    2017-07-01

    Plastid sequences are a cornerstone in plant systematic studies and key aspects of their evolution, such as uniparental inheritance and absent recombination, are often treated as axioms. While exceptions to these assumptions can profoundly influence evolutionary inference, detecting them can require extensive sampling, abundant sequence data, and detailed testing. Using advancements in high-throughput sequencing, we analyzed the whole plastomes of 65 accessions of Picea, a genus of ∼35 coniferous forest tree species, to test for deviations from canonical plastome evolution. Using complementary hypothesis and data-driven tests, we found evidence for chimeric plastomes generated by interspecific hybridization and recombination in the clade comprising Norway spruce (P. abies) and 10 other species. Support for interspecific recombination remained after controlling for sequence saturation, positive selection, and potential alignment artifacts. These results reconcile previous conflicting plastid-based phylogenies and strengthen the mounting evidence of reticulate evolution in Picea. Given the relatively high frequency of hybridization and biparental plastid inheritance in plants, we suggest interspecific plastome recombination may be more widespread than currently appreciated and could underlie reported cases of discordant plastid phylogenies. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  10. Ecological adaptations in Douglas-fir (Pseudotsuga menziesii var. glauca). IV. Montana and Idaho near the Continental Divide

    Science.gov (United States)

    Gerald Rehfeldt

    1988-01-01

    Seventy-seven seedling populations of Douglas-fir (Pseudotsuga menziesii var. glauca) sampled from near the Continental Divide in Idaho and Montana exhibited pronounced genetic differences when compared in three common environments. Differentiation involved several traits that are components of an annual developmental cycle that must be completed within a growing...

  11. Elevated UV-B radiation increased the decomposition of Cinnamomum camphora and Cyclobalanopsis glauca leaf litter in subtropical China

    Energy Technology Data Exchange (ETDEWEB)

    Song, Xinzhang Z.; Zhang, Huiling L.; Jiang, Hong; Yu, Shuquan Q. [Zhejiang Agriculture and Forestry Univ., Lin' an (China). The Nurturing Station for the State Key Lab. of Subtropical Silviculture; Zhejiang Agriculture and Forestry Univ., Lin' an (China). Zhejiang Provincial Key Lab. of Carbon Cycling and Carbon Sequestration in Forest Ecosystems; Chang, Scott X. [Alberta Univ., Edmonton (Canada). Dept. of Renewable Resources; Peng, Changhui H. [Quebec Univ., Montreal (Canada). Inst. of Environment Sciences

    2012-03-15

    Ultraviolet-B (UV-B) radiation reaching the earth's surface has been increasing due to ozone depletion and can profoundly influence litter decomposition and nutrient cycling in terrestrial ecosystems. The role of UV-B radiation in litter decomposition in humid environments is poorly understood; we thus investigated the effect of UV-B radiation on litter decomposition and nitrogen (N) release in a humid subtropical ecosystem in China. We conducted a field-based experiment using the litterbag method to study litter decomposition and N release under ambient and elevated (31% above ambient) UV-B radiation, using the leaf litter of two common tree species, Cinnamomum camphora and Cyclobalanopsis glauca, native to subtropical China. Elevated UV-B radiation significantly increased the decomposition rate of C. camphora and C. glauca leaf litter by 16.7% and 27.8%, respectively, and increased the N release from the decomposing litter of C. glauca but not C. camphora. Elevated UV-B radiation significantly accelerated the decomposition of litter of two native tree species and the N release from the decomposition litter of C. glauca in humid subtropical China, which has implications for soil carbon flux and forest productivity. (orig.)

  12. Size and distribution of Pandarus satyrus (Copepoda: Pandaridae) on the blue shark Prionace glauca (Carcharhiniformes: Carcharhinidae) in Costa Rica.

    Science.gov (United States)

    Rojas, J R; Rodríguez Solano, O; Morales-Ramírez, A

    2001-03-01

    A total of 80 specimens of Pandarus satyrus, a cosmopolitan ectoparasitic copepod, were taken from fishery catches of blue sharks (Prionace glauca) in the Eastern Pacific coast of Costa Rica. All specimens were found in the dorsal surface of pectoral fins (8-30 per shark). Longer specimens were most abundant.

  13. Construction of the BAC Library of Small Abalone (Haliotis diversicolor) for Gene Screening and Genome Characterization.

    Science.gov (United States)

    Jiang, Likun; You, Weiwei; Zhang, Xiaojun; Xu, Jian; Jiang, Yanliang; Wang, Kai; Zhao, Zixia; Chen, Baohua; Zhao, Yunfeng; Mahboob, Shahid; Al-Ghanim, Khalid A; Ke, Caihuan; Xu, Peng

    2016-02-01

    The small abalone (Haliotis diversicolor) is one of the most important aquaculture species in East Asia. To facilitate gene cloning and characterization, genome analysis, and genetic breeding of it, we constructed a large-insert bacterial artificial chromosome (BAC) library, which is an important genetic tool for advanced genetics and genomics research. The small abalone BAC library includes 92,610 clones with an average insert size of 120 Kb, equivalent to approximately 7.6× of the small abalone genome. We set up three-dimensional pools and super pools of 18,432 BAC clones for target gene screening using PCR method. To assess the approach, we screened 12 target genes in these 18,432 BAC clones and identified 16 positive BAC clones. Eight positive BAC clones were then sequenced and assembled with the next generation sequencing platform. The assembled contigs representing these 8 BAC clones spanned 928 Kb of the small abalone genome, providing the first batch of genome sequences for genome evaluation and characterization. The average GC content of small abalone genome was estimated as 40.33%. A total of 21 protein-coding genes, including 7 target genes, were annotated into the 8 BACs, which proved the feasibility of PCR screening approach with three-dimensional pools in small abalone BAC library. One hundred fifty microsatellite loci were also identified from the sequences for marker development in the future. The BAC library and clone pools provided valuable resources and tools for genetic breeding and conservation of H. diversicolor.

  14. Time to Detection with BacT/Alert FA Plus Compared to BacT/Alert FA Blood Culture Media.

    Science.gov (United States)

    Nutman, A; Fisher Even-Tsur, S; Shapiro, G; Braun, T; Schwartz, D; Carmeli, Y

    2016-09-01

    Rapid identification of the causative pathogen in patients with bacteremia allows adjustment of antibiotic therapy and improves patient outcomes. We compared in vitro and real-life time to detection (TTD) of two blood culture media, BacT/Alert FA (FA) and BacT/Alert FA Plus (FA Plus), for the nine most common species of bacterial pathogens recovered from blood samples. Experimental data from simulated cultures was compared with microbiology records of TTD for both culture media with growth of the species of interest in clinical blood cultures. In the experimental conditions, median TTD was 3.8 hours (23.9 %) shorter using FA Plus media. The magnitude of reduction differed between species. Similarly, in real life data, FA Plus had shorter TTD than FA media; however, the difference between culture media was smaller, and median TTD was only 1 hour (8.5 %) less. We found shorter TTD with BacT/Alert FA Plus culture media, both experimentally and in real-life conditions and unrelated to antibiotic neutralization, highlighting the importance of appropriate blood culture media selection.

  15. Partial Diversity Generates Effector Immunity Specificity of the Bac41-Like Bacteriocins of Enterococcus faecalis Clinical Strains.

    Science.gov (United States)

    Kurushima, Jun; Ike, Yasuyoshi; Tomita, Haruyoshi

    2016-09-01

    Bacteriocin 41 (Bac41) is the plasmid-encoded bacteriocin produced by the opportunistic pathogen Enterococcus faecalis Its genetic determinant consists of bacL1 (effector), bacL2 (regulator), bacA (effector), and bacI (immunity). The secreted effectors BacL1 and BacA coordinate to induce the lytic cell death of E. faecalis Meanwhile, the immunity factor BacI provides self-resistance to the Bac41 producer, E. faecalis, against the action of BacL1 and BacA. In this study, we demonstrated that more than half of the 327 clinical strains of E. faecalis screened had functional Bac41 genes. Analysis of the genetic structure of the Bac41 genes in the DNA sequences of the E. faecalis strains revealed that the Bac41-like genes consist of a relatively conserved region and a variable region located downstream from bacA Based on similarities in the variable region, the Bac41-like genes could be classified into type I, type IIa, and type IIb. Interestingly, the distinct Bac41 types had specific immunity factors for self-resistance, BacI1 or BacI2, and did not show cross-immunity to the other type of effector. We also demonstrated experimentally that the specificity of the immunity was determined by the combination of the C-terminal region of BacA and the presence of the unique BacI1 or BacI2 factor. These observations suggested that Bac41-like bacteriocin genes are extensively disseminated among E. faecalis strains in the clinical environment and can be grouped into at least three types. It was also indicated that the partial diversity results in specificity of self-resistance which may offer these strains a competitive advantage. Bacteriocins are antibacterial effectors produced by bacteria. In general, a bacteriocin-coding gene is accompanied by a cognate immunity gene that confers self-resistance on the bacteriocin-producing bacterium itself. We demonstrated that one of the bacteriocins, Bac41, is disseminated among E. faecalis clinical strains and the Bac41 subtypes with

  16. Complete mitochondrial genome of the blue shark Prionace glauca (Elasmobranchii: Carcharhiniformes).

    Science.gov (United States)

    Chen, Xiao; Xiang, Dan; Ai, Weiming; Shi, Xiaofang

    2015-04-01

    In this study, we first presented the complete mitochondrial genome of the blue shark Prionace Glauca, a pelagic and oceanic species. It is 16,705 bp in length and contains 2 rRNA genes, 22 tRNA genes, 13 protein-coding genes and 1 putative control region. The overall base composition is 31.6% A, 24.4% C, 13.1% G and 30.9% T. Overlaps and short inter-genic spaces are located in the genome. The tRNA-Ser2 loses the dihydrouridine arm and cannot be folded into the typical clover-leaf secondary structure. Two start codons (GTG and ATG) with two stop codons (TAG and TAA) or with one incomplete stop codon (T) are found in the 13 protein-coding genes. The control region contains high A + T (69.9%) and low G (12.0%).

  17. GenMapDB: a database of mapped human BAC clones

    OpenAIRE

    Morley, Michael; Arcaro, Melissa; Burdick, Joshua; Yonescu, Raluca; Reid, Thomas; Kirsch, Ilan R.; Cheung, Vivian G.

    2001-01-01

    GenMapDB (http://genomics.med.upenn.edu/genmapdb) is a repository of human bacterial artificial chromosome (BAC) clones mapped by our laboratory to sequence-tagged site markers. Currently, GenMapDB contains over 3000 mapped clones that span 19 chromosomes, chromosomes 2, 4, 5, 9–22, X and Y. This database provides positional information about human BAC clones from the RPCI-11 human male BAC library. It also contains restriction fragment analysis data and end sequen...

  18. Insulated piggyBac vectors for insect transgenesis

    Directory of Open Access Journals (Sweden)

    Horn Carsten

    2006-06-01

    Full Text Available Abstract Background Germ-line transformation of insects is now a widely used method for analyzing gene function and for the development of genetically modified strains suitable for pest control programs. The most widely used transposable element for the germ-line transformation of insects is piggyBac. The site of integration of the transgene can influence gene expression due to the effects of nearby transcription enhancers or silent heterochromatic regions. Position effects can be minimized by flanking a transgene with insulator elements. The scs/scs' and gypsy insulators from Drosophila melanogaster as well as the chicken β-globin HS4 insulator function in both Drosophila and mammalian cells. Results To minimize position effects we have created a set of piggyBac transformation vectors that contain either the scs/scs', gypsy or chicken β-globin HS4 insulators. The vectors contain either fluorescent protein or eye color marker genes and have been successfully used for germ-line transformation of Drosophila melanogaster. A set of the scs/scs' vectors contains the coral reef fluorescent protein marker genes AmCyan, ZsGreen and DsRed that have not been optimized for translation in human cells. These marker genes are controlled by a combined GMR-3xP3 enhancer/promoter that gives particularly strong expression in the eyes. This is also the first report of the use of the ZsGreen and AmCyan reef fluorescent proteins as transformation markers in insects. Conclusion The insulated piggyBac vectors should protect transgenes against position effects and thus facilitate fine control of gene expression in a wide spectrum of insect species. These vectors may also be used for transgenesis in other invertebrate species.

  19. SWFSC/MMTD/PI: Pacific Islands Cetacean Ecosystem Assessment Survey (PICEAS) 2005

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — PICEAS (Pacific Islands Cetacean Ecosystem Assessment Survey) 2005 was an ecosystem survey in the U.S. Exclusive Economic Zone (EEZ) waters of Palmyra and Johnston...

  20. A Blumeria graminis f.sp. hordei BAC library - contig building and microsynteny studies

    DEFF Research Database (Denmark)

    Pedersen, C.; Wu, B.; Giese, H.

    2002-01-01

    A bacterial artificial chromosome (BAC) library of Blumeria graminis f.sp. hordei, containing 12,000 clones with an average insert size of 41 kb, was constructed. The library represents about three genome equivalents and BAC-end sequencing showed a high content of repetitive sequences, making...... contigs, at or close to avirulence loci, were constructed. Single nucleotide polymorphism (SNP) markers were developed from BAC-end sequences to link the contigs to the genetic maps. Two other BAC contigs were used to study microsynteny between B. graminis and two other ascomycetes, Neurospora crassa...

  1. Lignin distribution in waterlogged archaeological Picea abies (L.) Karst degraded by erosion bacteria

    DEFF Research Database (Denmark)

    Pedersen, Nanna Bjerregaard; Schmitt, Uwe Schmitt; Koch, Gerald

    2014-01-01

    The lignin distribution in poles of waterlogged archaeological Picea abies (L.) Karst, which was decayed by erosion bacteria (EB) under anoxic conditions for approximately 400 years, was topochemically identified by transmission electron microscopy (TEM) and high resolution UV-microspectrophotome......The lignin distribution in poles of waterlogged archaeological Picea abies (L.) Karst, which was decayed by erosion bacteria (EB) under anoxic conditions for approximately 400 years, was topochemically identified by transmission electron microscopy (TEM) and high resolution UV...

  2. Size and distribution of Pandarus satyrus (Copepoda: Pandaridae on the blue shark Prionace glauca (Carcharhiniformes: Carcharhinidae in Costa Rica

    Directory of Open Access Journals (Sweden)

    José Rodrigo Rojas M.

    2001-03-01

    Full Text Available A total of 80 specimens of Pandarus satyrus, a cosmopolitan ectoparasitic copepod, were taken from fishery catches of blue sharks (Prionace glauca in the Eastern Pacific coast of Costa Rica. All specimens were found in the dorsal surface of pectoral fins (8-30 per shark. Longer specimens were most abundant.Un total de 80 especímenes de Pandarus satyrus, un copépodo ectoparásito cosmopolita, fueron tomados de tiburones azules (Prionace glauca capturados en la costa del Pacífico Oriental de Costa Rica. Todos los especímenes fueron encontrados en la superficie dorsal de las aletas pectorales (entre 8 y 30 por tiburón. Los parásitos más grandes son los más abundantes.

  3. THE DYNAMICS OF TOTAL AMYLASE\\'S ACTIVITY IN PANICUM MILIACEUM AND SETARIA GLAUCA DURING THE GERMINATION PERIOD

    Directory of Open Access Journals (Sweden)

    Elena Ciornea

    2006-08-01

    Full Text Available : It was studied the dynamics of total amylase\\'s activity in millet (Panicum miliaceum and bristle grass (Setaria glauca during the germination period. The enzymatic activity was determined by the Noelting - Brenfeld method, the results obtained being expressed in M maltose / g. In both millet and bristle grass, it was evidenced that both parameters taken into study (the species and the germination time do influence the enzymatic activity, although to a different extent.

  4. Pollination ecology of the invasive tree tobacco Nicotiana glauca: comparisons across native and non-native ranges

    Directory of Open Access Journals (Sweden)

    Jeff Ollerton

    2012-10-01

    Full Text Available Interactions with pollinators are thought to play a significant role in determining whether plant species become invasive, and ecologically generalised species are predicted to be more likely to invade than more specialised species. Using published and unpublished data we assessed the floral biology and pollination ecology of the South American native Nicotiana glauca (Solanaceae which has become a significant invasive of semi-arid parts of the world. In regions where specialised bird pollinators are available, for example hummingbirds in California and sunbirds in South Africa and Israel, N. glauca interacts with these local pollinators and sets seed by both out-crossing and selfing. In areas where there are no such birds, such as the Canary Islands and Greece, abundant viable seed is set by selfing, facilitated by the shorter stigma-anther distance compared to plants in native populations. Surprisingly, in these areas without pollinating birds, the considerable nectar resources are only rarely exploited by other flower visitors such as bees or butterflies, either legitimately or by nectar robbing. We conclude that Nicotiana glauca is a successful invasive species outside of its native range, despite its functionally specialised hummingbird pollination system, because it has evolved to become more frequently self pollinating in areas where it is introduced. Its invasion success is not predictable from what is known of its interactions with pollinators in its home range.

  5. [Compatible biomass models of natural spruce (Picea asperata)].

    Science.gov (United States)

    Wang, Jin Chi; Deng, Hua Feng; Huang, Guo Sheng; Wang, Xue Jun; Zhang, Lu

    2017-10-01

    By using nonlinear measurement error method, the compatible tree volume and above ground biomass equations were established based on the volume and biomass data of 150 sampling trees of natural spruce (Picea asperata). Two approaches, controlling directly under total aboveground biomass and controlling jointly from level to level, were used to design the compatible system for the total aboveground biomass and the biomass of four components (stem, bark, branch and foliage), and the total ground biomass could be estimated independently or estimated simultaneously in the system. The results showed that the R 2 of the one variable and bivariate compatible tree volume and aboveground biomass equations were all above 0.85, and the maximum value reached 0.99. The prediction effect of the volume equations could be improved significantly when tree height was included as predictor, while it was not significant in biomass estimation. For the compatible biomass systems, the one variable model based on controlling jointly from level to level was better than the model using controlling directly under total above ground biomass, but the bivariate models of the two methods were similar. Comparing the imitative effects of the one variable and bivariate compatible biomass models, the results showed that the increase of explainable variables could significantly improve the fitness of branch and foliage biomass, but had little effect on other components. Besides, there was almost no difference between the two methods of estimation based on the comparison.

  6. Recombineering strategies for developing next generation BAC transgenic tools for optogenetics and beyond.

    Science.gov (United States)

    Ting, Jonathan T; Feng, Guoping

    2014-01-01

    The development and application of diverse BAC transgenic rodent lines has enabled rapid progress for precise molecular targeting of genetically-defined cell types in the mammalian central nervous system. These transgenic tools have played a central role in the optogenetic revolution in neuroscience. Indeed, an overwhelming proportion of studies in this field have made use of BAC transgenic Cre driver lines to achieve targeted expression of optogenetic probes in the brain. In addition, several BAC transgenic mouse lines have been established for direct cell-type specific expression of Channelrhodopsin-2 (ChR2). While the benefits of these new tools largely outweigh any accompanying challenges, many available BAC transgenic lines may suffer from confounds due in part to increased gene dosage of one or more "extra" genes contained within the large BAC DNA sequences. Here we discuss this under-appreciated issue and propose strategies for developing the next generation of BAC transgenic lines that are devoid of extra genes. Furthermore, we provide evidence that these strategies are simple, reproducible, and do not disrupt the intended cell-type specific transgene expression patterns for several distinct BAC clones. These strategies may be widely implemented for improved BAC transgenesis across diverse disciplines.

  7. Effects of nitrogen ion irradiation on endoglucanase activity and gene mutation of Bacillus subtilis Bac01

    International Nuclear Information System (INIS)

    Lv Jie; Mao Peihong; Jin Xiang; Yu Long; Ying Hanjie

    2009-01-01

    Bacillus subtilis Bac01 was mutated by 15 keV N + ions of 1.5xl0 16 cm -2 . The mutant strain Bac11 with high yield of endoglucanase was isolated using carboxymethylcellulose sodium and congo red indicative plates. It exhibited higher endoglucanase activity (381.89IU) than the original strain Bac01 (93.33IU). Two 1,500 bp endoglucanase gene fragments were obtained with PCR amplification from B. subtilis Bac01 and mutant strain Bac11. BLAST comparison result indicated that 10 nucleotides mutated. Bioinformatics methods were used to analyze the two predicted amino acid sequences, and it was found that 5 amino acid residues changed, being all in the cellulose-binding domain of endoglucanase. (authors)

  8. Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens

    KAUST Repository

    Ali, Shawkat

    2014-09-19

    The genetic transformation of certain organisms, required for gene function analysis or complementation, is often not very efficient, especially when dealing with large gene constructs or genomic fragments. We have adapted the natural DNA transfer mechanism from the soil pathogenic bacterium Agrobacterium tumefaciens, to deliver intact large DNA constructs to basidiomycete fungi of the genus Ustilago where they stably integrated into their genome. To this end, Bacterial Artificial Chromosome (BAC) clones containing large fungal genomic DNA fragments were converted via a Lambda phage-based recombineering step to Agrobacterium transfer-competent binary vectors (BIBACs) with a Ustilago-specific selection marker. The fungal genomic DNA fragment was subsequently successfully delivered as T-DNA through Agrobacterium-mediated transformation into Ustilago species where an intact copy stably integrated into the genome. By modifying the recombineering vector, this method can theoretically be adapted for many different fungi.

  9. A Comparative BAC Map for the Gilthead Sea Bream (Sparus aurata L.

    Directory of Open Access Journals (Sweden)

    Heiner Kuhl

    2011-01-01

    Full Text Available This study presents the first comparative BAC map of the gilthead sea bream (Sparus aurata, a highly valuated marine aquaculture fish species in the Mediterranean. High-throughput end sequencing of a BAC library yielded 92,468 reads (60.6 Mbp. Comparative mapping was achieved by anchoring BAC end sequences to the three-spined stickleback (Gasterosteus aculeatus genome. BACs that were consistently ordered along the stickleback chromosomes accounted for 14,265 clones. A fraction of 5,249 BACs constituted a minimal tiling path that covers 73.5% of the stickleback chromosomes and 70.2% of the genes that have been annotated. The N50 size of 1,485 “BACtigs” consisting of redundant BACs is 337,253 bp. The largest BACtig covers 2.15 Mbp in the stickleback genome. According to the insert size distribution of mapped BACs the sea bream genome is 1.71-fold larger than the stickleback genome. These results represent a valuable tool to researchers in the field and may support future projects to elucidate the whole sea bream genome.

  10. Sensitivity and applicability of the Brazilian version of the Brief Assessment of Cognition in Schizophrenia (BACS

    Directory of Open Access Journals (Sweden)

    João Vinícius Salgado

    Full Text Available Abstract Cognitive assessment in schizophrenia has traditionally used batteries that are long and complex or differ widely in their content. The Brief Assessment of Cognition in Schizophrenia (BACS has been developed to cover the main cognitive deficits of schizophrenia as well as to be easily and briefly administered, portable, sensitive and reliable. Objectives: To investigate the applicability and sensitivity of the Brazilian Version of the BACS (Brazilian-BACS. Methods: Performance of 20 stable patients with schizophrenia on the Brazilian-BACS was compared to 20 matched healthy controls. Results: Applying the Brazilian-BACS required 43.4±8.4 minutes for patients and 40.5±5.7 minutes for controls (p=0.17. All tests demonstrated significant differences between controls and patients (P<0.01. Pearson's correlation analysis and Cronbach's a evidenced a high internal consistency for patient performance. The cognitive deficit in the patients was approximately 1.5 standard deviations below controls. These results were consistent with those reported in the validation of the original version and in meta-analyses of similar studies. Conclusions: The Brazilian-BACS displayed good applicability and sensitivity in assessing the major cognitive constructs that are impaired in schizophrenia. Thus, the Brazilian-BACS seems to be a promising tool for assessing cognition in patients with schizophrenia in Brazil.

  11. A controlled comparison of the BacT/ALERT® 3D and VIRTUO™ microbial detection systems.

    Science.gov (United States)

    Totty, H; Ullery, M; Spontak, J; Viray, J; Adamik, M; Katzin, B; Dunne, W M; Deol, P

    2017-10-01

    The performance of the next-generation BacT/ALERT® VIRTUO™ Microbial Detection System (VIRTUO™, bioMérieux Inc., Hazelwood, MO) was compared to the BacT/ALERT® 3D Microbial Detection System (3D, bioMérieux Inc., Durham, NC) using BacT/ALERT® FA Plus (FA Plus), BacT/ALERT® PF Plus (PF Plus), BacT/ALERT® FN Plus (FN Plus), BacT/ALERT® Standard Aerobic (SA), and BacT/ALERT® Standard Anaerobic (SN) blood culture bottles (bioMérieux Inc., Durham, NC). A seeded limit of detection (LoD) study was performed for each bottle type in both systems. The LoD studies demonstrated that both systems were capable of detecting organisms at nearly identical levels [detection (TTD) between the systems using a panel of clinically relevant microorganisms inoculated at or near the LoD with 0, 4, or 10 mL of healthy human blood. VIRTUO™ exhibited a faster TTD by an average of 3.5 h, as well as demonstrated a significantly improved detection rate of 99.9% compared to 98.8% with 3D (p-value <0.05).

  12. Toward an Integrated BAC Library Resource for Genome Sequencing and Analysis; FINAL

    International Nuclear Information System (INIS)

    Simon, M. I.; Kim, U.-J.

    2002-01-01

    We developed a great deal of expertise in building large BAC libraries from a variety of DNA sources including humans, mice, corn, microorganisms, worms, and Arabidopsis. We greatly improved the technology for screening these libraries rapidly and for selecting appropriate BACs and mapping BACs to develop large overlapping contigs. We became involved in supplying BACs and BAC contigs to a variety of sequencing and mapping projects and we began to collaborate with Drs. Adams and Venter at TIGR and with Dr. Leroy Hood and his group at University of Washington to provide BACs for end sequencing and for mapping and sequencing of large fragments of chromosome 16. Together with Dr. Ian Dunham and his co-workers at the Sanger Center we completed the mapping and they completed the sequencing of the first human chromosome, chromosome 22. This was published in Nature in 1999 and our BAC contigs made a major contribution to this sequencing effort. Drs. Shizuya and Ding invented an automated highly accurate BAC mapping technique. We also developed long-term collaborations with Dr. Uli Weier at UCSF in the design of BAC probes for characterization of human tumors and specific chromosome deletions and breakpoints. Finally the contribution of our work to the human genome project has been recognized in the publication both by the international consortium and the NIH of a draft sequence of the human genome in Nature last year. Dr. Shizuya was acknowledged in the authorship of that landmark paper. Dr. Simon was also an author on the Venter/Adams Celera project sequencing the human genome that was published in Science last year

  13. Lophodermium piceae and Tryblidiopsis pinastri. Two latent colonizers of Norway spruce

    Energy Technology Data Exchange (ETDEWEB)

    Lehtijaervi, A. [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Forest Mycology and Pathology

    1998-12-31

    Among the endophytic microfungi colonizing Norway spruce, the non-pathogenic species Lophodermium piceae and Tryblidiopsis pinastri are ubiquitous. Most Norway spruce in Sweden are colonized by these fungi. L. piceae colonizes healthy needles of various ages, while T pinastri colonizes the bark of branches. New shoots become infected around the time of their emergence. Fruit bodies are formed after the needles and branches die. It was found that L. piceae colonized needles during the summer and early autumn, about six weeks after a prolonged period of intensive rain. The number of individual fungal mycelia per needle increased linearly with needle age. In experiments with trees, irrigation resulted in lower total colonization and delayed colonization of the current-year needles, regardless of whether fertilizer had been added. Colonization by L. piceae was lowest in the ammonium sulfate treatment, which simulated the deposition of air pollutants. Colonization was highest in the control and drought treatments, which did not differ significantly from each other. Only a few pathogenic fungi can damage Norway spruce needles. Needles damaged by the rust fungus Chrysomyxa abietis were investigated to study interactions between fungi. The frequency and intensity of L. piceae colonization were found to be similar for C. abietis infected and healthy needles. However, in needles partially infected with rust, L. piceae seemed to establish itself easier in the rust-infected part than in the green part. The genetic structures of populations of T. pinastri in southern Sweden and Finland were investigated using DNA markers produced by means of arbitrarily primed PCR. Single spore isolates from apothecia were used in the analysis. A considerable amount of variation was detected. No geographical differentiation was found among the populations studied

  14. Observation on the effect of gamma rays on spruce (Picea smithiana) seeds germination

    International Nuclear Information System (INIS)

    Sharma, S.K.

    1997-01-01

    Spruce (Picea smithiana) forests cover extensive areas in western Himalayas and account for 48.80% of the total conifer forest area and growing stock in the region. Baldwin studied the effect of x-rays on the seeds of conifers trees. A little work has been done on the genus Picea in respect of physical mutagens. Keeping this in consideration the present trial has been done at preliminary level to study the effect of gamma rays on the germination behaviour. (author). 3 refs., 1 tab

  15. Registro de un Monstruo Diprósopo de Tiburón Azul (Prionace glauca) en Chile

    OpenAIRE

    Hevia-Hormazábal, Valentina; Pastén-Marambio, Víctor; Vega, Alonso

    2011-01-01

    Un espécimen de siameses de tiburón azul, Prionace glauca, fue recolectado durante actividades de pesca en el norte de Chile. Según una clasificación basada en el grado de duplicación y el sitio de fusión de los gemelos, el espécimen corresponde a un monstruo del tipo diprósopo tetroftalmo. El individuo tiene una duplicación craneofacial parcial con presencia de cuatro globos oculares e igual número de narinas. Las cavidades orales están interconectadas convergiendo en una única faringe. El t...

  16. Integrated analysis of 454 and Illumina transcriptomic sequencing characterizes carbon flux and energy source for fatty acid synthesis in developing Lindera glauca fruits for woody biodiesel.

    Science.gov (United States)

    Lin, Zixin; An, Jiyong; Wang, Jia; Niu, Jun; Ma, Chao; Wang, Libing; Yuan, Guanshen; Shi, Lingling; Liu, Lili; Zhang, Jinsong; Zhang, Zhixiang; Qi, Ji; Lin, Shanzhi

    2017-01-01

    Lindera glauca fruit with high quality and quantity of oil has emerged as a novel potential source of biodiesel in China, but the molecular regulatory mechanism of carbon flux and energy source for oil biosynthesis in developing fruits is still unknown. To better develop fruit oils of L. glauca as woody biodiesel, a combination of two different sequencing platforms (454 and Illumina) and qRT-PCR analysis was used to define a minimal reference transcriptome of developing L. glauca fruits, and to construct carbon and energy metabolic model for regulation of carbon partitioning and energy supply for FA biosynthesis and oil accumulation. We first analyzed the dynamic patterns of growth tendency, oil content, FA compositions, biodiesel properties, and the contents of ATP and pyridine nucleotide of L. glauca fruits from seven different developing stages. Comprehensive characterization of transcriptome of the developing L. glauca fruit was performed using a combination of two different next-generation sequencing platforms, of which three representative fruit samples (50, 125, and 150 DAF) and one mixed sample from seven developing stages were selected for Illumina and 454 sequencing, respectively. The unigenes separately obtained from long and short reads (201, and 259, respectively, in total) were reconciled using TGICL software, resulting in a total of 60,031 unigenes (mean length = 1061.95 bp) to describe a transcriptome for developing L. glauca fruits. Notably, 198 genes were annotated for photosynthesis, sucrose cleavage, carbon allocation, metabolite transport, acetyl-CoA formation, oil synthesis, and energy metabolism, among which some specific transporters, transcription factors, and enzymes were identified to be implicated in carbon partitioning and energy source for oil synthesis by an integrated analysis of transcriptomic sequencing and qRT-PCR. Importantly, the carbon and energy metabolic model was well established for oil biosynthesis of developing L

  17. BAC-Dkk3-EGFP Transgenic Mouse: An In Vivo Analytical Tool for Dkk3 Expression

    Directory of Open Access Journals (Sweden)

    Yuki Muranishi

    2012-01-01

    Full Text Available Dickkopf (DKK family proteins are secreted modulators of the Wnt signaling pathway and are capable of regulating the development of many organs and tissues. We previously identified Dkk3 to be a molecule predominantly expressed in the mouse embryonic retina. However, which cell expresses Dkk3 in the developing and mature mouse retina remains to be elucidated. To examine the precise expression of the Dkk3 protein, we generated BAC-Dkk3-EGFP transgenic mice that express EGFP integrated into the Dkk3 gene in a BAC plasmid. Expression analysis using the BAC-Dkk3-EGFP transgenic mice revealed that Dkk3 is expressed in retinal progenitor cells (RPCs at embryonic stages and in Müller glial cells in the adult retina. Since Müller glial cells may play a potential role in retinal regeneration, BAC-Dkk3-EGFP mice could be useful for retinal regeneration studies.

  18. Expression Analysis of CB2-GFP BAC Transgenic Mice.

    Science.gov (United States)

    Schmöle, Anne-Caroline; Lundt, Ramona; Gennequin, Benjamin; Schrage, Hanna; Beins, Eva; Krämer, Alexandra; Zimmer, Till; Limmer, Andreas; Zimmer, Andreas; Otte, David-Marian

    2015-01-01

    The endocannabinoid system (ECS) is a retrograde messenger system, consisting of lipid signaling molecules that bind to at least two G-protein-coupled receptors, Cannabinoid receptor 1 and 2 (CB1 and 2). As CB2 is primarily expressed on immune cells such as B cells, T cells, macrophages, dendritic cells, and microglia, it is of great interest how CB2 contributes to immune cell development and function in health and disease. Here, understanding the mechanisms of CB2 involvement in immune-cell function as well as the trafficking and regulation of CB2 expressing cells are crucial issues. Up to now, CB2 antibodies produce unclear results, especially those targeting the murine protein. Therefore, we have generated BAC transgenic GFP reporter mice (CB2-GFPTg) to trace CB2 expression in vitro and in situ. Those mice express GFP under the CB2 promoter and display GFP expression paralleling CB2 expression on the transcript level in spleen, thymus and brain tissue. Furthermore, by using fluorescence techniques we show that the major sources for GFP-CB2 expression are B cells in spleen and blood and microglia in the brain. This novel CB2-GFP transgenic reporter mouse line represents a powerful resource to study CB2 expression in different cell types. Furthermore, it could be used for analyzing CB2-mediated mobilization and trafficking of immune cells as well as studying the fate of recruited immune cells in models of acute and chronic inflammation.

  19. Expression Analysis of CB2-GFP BAC Transgenic Mice.

    Directory of Open Access Journals (Sweden)

    Anne-Caroline Schmöle

    Full Text Available The endocannabinoid system (ECS is a retrograde messenger system, consisting of lipid signaling molecules that bind to at least two G-protein-coupled receptors, Cannabinoid receptor 1 and 2 (CB1 and 2. As CB2 is primarily expressed on immune cells such as B cells, T cells, macrophages, dendritic cells, and microglia, it is of great interest how CB2 contributes to immune cell development and function in health and disease. Here, understanding the mechanisms of CB2 involvement in immune-cell function as well as the trafficking and regulation of CB2 expressing cells are crucial issues. Up to now, CB2 antibodies produce unclear results, especially those targeting the murine protein. Therefore, we have generated BAC transgenic GFP reporter mice (CB2-GFPTg to trace CB2 expression in vitro and in situ. Those mice express GFP under the CB2 promoter and display GFP expression paralleling CB2 expression on the transcript level in spleen, thymus and brain tissue. Furthermore, by using fluorescence techniques we show that the major sources for GFP-CB2 expression are B cells in spleen and blood and microglia in the brain. This novel CB2-GFP transgenic reporter mouse line represents a powerful resource to study CB2 expression in different cell types. Furthermore, it could be used for analyzing CB2-mediated mobilization and trafficking of immune cells as well as studying the fate of recruited immune cells in models of acute and chronic inflammation.

  20. Uncertainty of Blood Alcohol Concentration (BAC Results as Related to Instrumental Conditions: Optimization and Robustness of BAC Analysis Headspace Parameters

    Directory of Open Access Journals (Sweden)

    Haleigh A. Boswell

    2015-12-01

    Full Text Available Analysis of blood alcohol concentration is a routine analysis performed in many forensic laboratories. This analysis commonly utilizes static headspace sampling, followed by gas chromatography combined with flame ionization detection (GC-FID. Studies have shown several “optimal” methods for instrumental operating conditions, which are intended to yield accurate and precise data. Given that different instruments, sampling methods, application specific columns and parameters are often utilized, it is much less common to find information on the robustness of these reported conditions. A major problem can arise when these “optimal” conditions may not also be robust, thus producing data with higher than desired uncertainty or potentially inaccurate results. The goal of this research was to incorporate the principles of quality by design (QBD in the adjustment and determination of BAC (blood alcohol concentration instrumental headspace parameters, thereby ensuring that minor instrumental variations, which occur as a matter of normal work, do not appreciably affect the final results of this analysis. This study discusses both the QBD principles as well as the results of the experiments, which allow for determination of more favorable instrumental headspace conditions. Additionally, method detection limits will also be reported in order to determine a reporting threshold and the degree of uncertainty at the common threshold value of 0.08 g/dL. Furthermore, the comparison of two internal standards, n-propanol and t-butanol, will be investigated. The study showed that an altered parameter of 85 °C headspace oven temperature and 15 psi headspace vial pressurization produces the lowest percent relative standard deviation of 1.3% when t-butanol is implemented as an internal standard, at least for one very common platform. The study also showed that an altered parameter of 100 °C headspace oven temperature and 15-psi headspace vial pressurization

  1. BAC-HAPPY mapping (BAP mapping: a new and efficient protocol for physical mapping.

    Directory of Open Access Journals (Sweden)

    Giang T H Vu

    2010-02-01

    Full Text Available Physical and linkage mapping underpin efforts to sequence and characterize the genomes of eukaryotic organisms by providing a skeleton framework for whole genome assembly. Hitherto, linkage and physical "contig" maps were generated independently prior to merging. Here, we develop a new and easy method, BAC HAPPY MAPPING (BAP mapping, that utilizes BAC library pools as a HAPPY mapping panel together with an Mbp-sized DNA panel to integrate the linkage and physical mapping efforts into one pipeline. Using Arabidopsis thaliana as an exemplar, a set of 40 Sequence Tagged Site (STS markers spanning approximately 10% of chromosome 4 were simultaneously assembled onto a BAP map compiled using both a series of BAC pools each comprising 0.7x genome coverage and dilute (0.7x genome samples of sheared genomic DNA. The resultant BAP map overcomes the need for polymorphic loci to separate genetic loci by recombination and allows physical mapping in segments of suppressed recombination that are difficult to analyze using traditional mapping techniques. Even virtual "BAC-HAPPY-mapping" to convert BAC landing data into BAC linkage contigs is possible.

  2. The Functionality of Minimal PiggyBac Transposons in Mammalian Cells

    Directory of Open Access Journals (Sweden)

    Boris Troyanovsky

    2016-01-01

    Full Text Available Minimal piggyBac vectors are a modified single-plasmid version of the classical piggyBac delivery system that can be used for stable transgene integration. These vectors have a truncated terminal domain in the delivery cassette and thus, integrate significantly less flanking transposon DNA into host cell chromatin than classical piggyBac vectors. Herein, we test various characteristics of this modified transposon. The integration efficiency of minimal piggyBac vectors was inversely related to the size of both the transposon and the entire plasmid, but inserts as large as 15 kb were efficiently integrated. Open and super-coiled vectors demonstrated the same integration efficiency while DNA methylation decreased the integration efficiency and silenced the expression of previously integrated sequences in some cell types. Importantly, the incidence of plasmid backbone integration was not increased above that seen in nontransposon control vectors. In BALB/c mice, we demonstrated prolonged expression of two transgenes (intracellular mCherry and secretable Gaussia luciferase when delivered by the minimal piggyBac that resulted in a more sustained antibody production against the immunogenic luciferase than when delivered by a transient (nontransposon vector plasmid. We conclude that minimal piggyBac vectors are an effective alternative to other integrative systems for stable DNA delivery in vitro and in vivo.

  3. Sequencing of BAC pools by different next generation sequencing platforms and strategies

    Directory of Open Access Journals (Sweden)

    Scholz Uwe

    2011-10-01

    Full Text Available Abstract Background Next generation sequencing of BACs is a viable option for deciphering the sequence of even large and highly repetitive genomes. In order to optimize this strategy, we examined the influence of read length on the quality of Roche/454 sequence assemblies, to what extent Illumina/Solexa mate pairs (MPs improve the assemblies by scaffolding and whether barcoding of BACs is dispensable. Results Sequencing four BACs with both FLX and Titanium technologies revealed similar sequencing accuracy, but showed that the longer Titanium reads produce considerably less misassemblies and gaps. The 454 assemblies of 96 barcoded BACs were improved by scaffolding 79% of the total contig length with MPs from a non-barcoded library. Assembly of the unmasked 454 sequences without separation by barcodes revealed chimeric contig formation to be a major problem, encompassing 47% of the total contig length. Masking the sequences reduced this fraction to 24%. Conclusion Optimal BAC pool sequencing should be based on the longest available reads, with barcoding essential for a comprehensive assessment of both repetitive and non-repetitive sequence information. When interest is restricted to non-repetitive regions and repeats are masked prior to assembly, barcoding is non-essential. In any case, the assemblies can be improved considerably by scaffolding with non-barcoded BAC pool MPs.

  4. Exogenous putrescine affects endogenous polyamine levels and the development of Picea abies somatic embryos

    Czech Academy of Sciences Publication Activity Database

    Vondráková, Zuzana; Eliášová, Kateřina; Vágner, Martin; Martincová, Olga; Cvikrová, Milena

    2015-01-01

    Roč. 75, č. 2 (2015), s. 405-414 ISSN 0167-6903 R&D Projects: GA MŠk(CZ) LD13050 Institutional support: RVO:61389030 Keywords : Exogenous putrescine * Somatic embryogenesis * Picea abies Subject RIV: ED - Physiology Impact factor: 2.333, year: 2015

  5. Winter photosynthesis in red spruce (Picea rubens Sarg.): limitations, potential benefits, and risks

    Science.gov (United States)

    P.G. Schaberg

    2000-01-01

    Numerous cold-induced changes in physiology limit the capacity of northern conifers to photosynthesize during winter. Studies of red spruce (Picea rubens Sarg.) have shown that rates of field photosynthesis (Pfield) and laboratory measurements of photosynthetic capacity (Pmax) generally parallel seasonal...

  6. Indicators of Population Viabllity in Red Spruce, Picea rubens. I. Reproductive Traits and Fecundity

    Science.gov (United States)

    A. Mosseler; J.E. Major; J.D. Simpson; B. Daigle; K. Lange; Y.S. Park; K.H Johnsen; O.P. Rajora

    2000-01-01

    Red spruce (Picea rubens Sarg.) has experienced a substantial decline across most of its range in eastern North America over the past centmy and probably also in the disjunct Ontario populations where it now occurs only in small isolated stands. Measurements of cone and seed traits from natural populations were used as indicators of the...

  7. Norway spruce (Picea abies) genetic transformation with modified Cry3A gene of Bacillus thuringiensis

    Czech Academy of Sciences Publication Activity Database

    Bříza, Jindřich; Pavingerová, Daniela; Vlasák, Josef; Niedermeierová, Hana

    2013-01-01

    Roč. 60, č. 3 (2013), s. 395-400 ISSN 0001-527X R&D Projects: GA MZe QH71290; GA ČR(CZ) GAP502/11/1471 Institutional support: RVO:60077344 Keywords : Cry3A gene modification * Picea abies * Agrobacterium tumefaciens Subject RIV: EB - Genetic s ; Molecular Biology Impact factor: 1.389, year: 2013

  8. Genetic diversity and conservation of Picea chihuahuana Martínez ...

    African Journals Online (AJOL)

    The conservation of genetic diversity in tree populations is an essential component of sustainable forest management. Picea chihuahuana Martínez is an endemic conifer species in Mexico and is considered to be endangered. P. chihuahuana covers a total area of no more than 300 ha at the Sierra Madre Occidental, ...

  9. Complex bud architecture and cell-specific chemical patterns enable supercooling of Picea abies bud primordial

    Science.gov (United States)

    Bud primordia of Picea abies, despite a frozen shoot, stay ice free down to -50 °C by a mechanism termed supercooling whose biophysical and biochemical requirements are poorly understood. Bud architecture was assessed by 3D-reconstruction, supercooling and freezing patterns by infrared video thermog...

  10. Germ-line transformation of the Queensland fruit fly, Bactrocera tryoni, using a piggyBac vector in the presence of endogenous piggyBac elements

    Science.gov (United States)

    We report the stable genetic transformation of the Queensland fruit fly Bactrocera tryoni using a piggyBac vector marked with either the fluorescent protein DsRed or EGFP.A transformation frequency of 5–10% was obtained.Inheritance of the transgenes has remained stable over eight generations despite...

  11. Bacterial contamination of platelet components not detected by BacT/ALERT®.

    Science.gov (United States)

    Abela, M A; Fenning, S; Maguire, K A; Morris, K G

    2018-02-01

    To investigate the possible causes for false negative results in BacT/ALERT ® 3D Signature System despite bacterial contamination of platelet units. The Northern Ireland Blood Transfusion Service (NIBTS) routinely extends platelet component shelf life to 7 days. Components are sampled and screened for bacterial contamination using an automated microbial detection system, the BacT/ALERT ® 3D Signature System. We report on three platelet components with confirmed bacterial contamination, which represent false negative BacT/ALERT ® results and near-miss serious adverse events. NIBTS protocols for risk reduction of bacterial contamination of platelet components are described. The methodology for bacterial detection using BacT/ALERT ® is outlined. Laboratory tests, relevant patient details and relevant follow-up information are analysed. In all three cases, Staphylococcus aureus was isolated from the platelet residue and confirmed on terminal sub-culture using BacT/ALERT ® . In two cases, S. aureus with similar genetic makeup was isolated from the donors. Risk reduction measures for bacterial contamination of platelet components are not always effective. Automated bacterial culture detection does not eliminate the risk of bacterial contamination. Visual inspection of platelet components prior to release, issue and administration remains an important last line of defence. © 2017 British Blood Transfusion Society.

  12. Evaluation of the present genetic conservation efforts in Pinus sylvestris, Picea abies, Quercus spp., Fagus sylvatica, and Pinus pinaster

    NARCIS (Netherlands)

    Kramer, K.

    2015-01-01

    Information on genetic diversity and gene conservation activities were combined with climatic data to evaluate the present genetic conservation efforts in Pinus sylvestris, Picea abies, Quercus spp., Fagus sylvatica, and Pinus pinaster. Combinations of climatic variables explained much of the

  13. Polychlorinated biphenyls and DDT in swordfish (Xiphias gladius) and blue shark (Prionace glauca) from Brazilian Coast. Preliminary results

    Energy Technology Data Exchange (ETDEWEB)

    Azevedo e Silva, C E; Azeredo, A; Meire, R; Torres, J P [Laboratorio de Radioisotopos E.P.F., Rio de Janeiro (Brazil). Inst. de Biofisica Carlos Chagas Filho, UFRJ; Brito, Jr, J L; Malm, O [Projeto Mamiferos Aquaticos, Rio de Janeiro (Brazil). Dept. de Oceanografia, UERJ

    2004-09-15

    There is conclusive evidence showing that, in general, fish in meals human diet contributes with a significant proportion of the total intake of PCBs and others organochlorine compounds, particularly fish with higher fat content. Thus, human exposure to PCBs is predominantly via diet, and especially from fish and seafood products. Comparatively, little is know about organochlorine contaminants in elasmobranch species, although they are also top predators. Characteristically, sharks are live longer, with comparatively slow rates of growth that in conjuction with their high trophic position may contribute to the accumulation of high concentrations of pollutants. Blue shark and swordfish are carnivorous fishes of great economic importance. This study investigate the concentration of polychlorinated biphenyls (PCBs) and DDTs in samples of muscle tissues of blue shark (Prionace glauca) and swordfish (Xiphias gladius) from Brazilian Coast.

  14. The European sea bass Dicentrarchus labrax genome puzzle: comparative BAC-mapping and low coverage shotgun sequencing

    Directory of Open Access Journals (Sweden)

    Volckaert Filip AM

    2010-01-01

    Full Text Available Abstract Background Food supply from the ocean is constrained by the shortage of domesticated and selected fish. Development of genomic models of economically important fishes should assist with the removal of this bottleneck. European sea bass Dicentrarchus labrax L. (Moronidae, Perciformes, Teleostei is one of the most important fishes in European marine aquaculture; growing genomic resources put it on its way to serve as an economic model. Results End sequencing of a sea bass genomic BAC-library enabled the comparative mapping of the sea bass genome using the three-spined stickleback Gasterosteus aculeatus genome as a reference. BAC-end sequences (102,690 were aligned to the stickleback genome. The number of mappable BACs was improved using a two-fold coverage WGS dataset of sea bass resulting in a comparative BAC-map covering 87% of stickleback chromosomes with 588 BAC-contigs. The minimum size of 83 contigs covering 50% of the reference was 1.2 Mbp; the largest BAC-contig comprised 8.86 Mbp. More than 22,000 BAC-clones aligned with both ends to the reference genome. Intra-chromosomal rearrangements between sea bass and stickleback were identified. Size distributions of mapped BACs were used to calculate that the genome of sea bass may be only 1.3 fold larger than the 460 Mbp stickleback genome. Conclusions The BAC map is used for sequencing single BACs or BAC-pools covering defined genomic entities by second generation sequencing technologies. Together with the WGS dataset it initiates a sea bass genome sequencing project. This will allow the quantification of polymorphisms through resequencing, which is important for selecting highly performing domesticated fish.

  15. [Pathways and rates of Pinus sylvestris L. and Picea species recolonization into Scandinavia in Holocene].

    Science.gov (United States)

    Sannikov, S N; Sannikova, N S

    2015-01-01

    The results are presented of comparative analysis of pathways, rates, and timing of recolonization into Scandinavia, in Holocene, of Pinus sylvestris populations and those of Picea abies and P. obovata. The dispersion rate, starting from 12 thou years before present (BP), is calculated using palynological data from scientific literature on radiometric dating. It is found out that P sylvestris spread into Central Scandinavia from the Alps via the Danish Isthmus about 8.2 thou years BP with the speed of 500-1250 km per 1 thou years. A hypothesis is put forward suggesting that such a fast speed is due to pine seeds hydrochory, which is much faster than anemochory according to our researches. From the northern part of the East European Plain, P. sylvestris spread into Fennoscandia with lower speed (520 km per 1 thou years). Populations of Picea species dispersed from the same regions with speed (131-164 km per 1 thou years) 3-10 times lower than that of P. sylvestris. Therefore, invasion of Picea abies from the Alps into Scandinavia via the Danish Isthmus did not have time to happen before the formation of the Kattegat Strait. By circumferential pathway, through Karelia, both species of Picea reached the northern parts of Scandinavia only 3.5 thou years BP, its central parts - 2 thou years BP, and its southern parts - 1.5 thou years BP, i.e., later than P. sylvestris by 4, 6.2, and 8.5 thou years respectively. Probably, this may be explained by the fact that in pines the time to seeding is twofold shorter, while their sprouts were more tolerant to climatic extremums in periglacial habitats in middle Holocene.

  16. Modelling Age- and Density-Related Gas Exchange of Picea abies Canopies in the Fichtelgebirge, Germany

    OpenAIRE

    Falge, Eva; Tennhunen, John D.; Ryel, Ronald J.; Alsheimer, Martina; Köstner, Barbara

    2000-01-01

    International audience; Differences in canopy exchange of water and carbon dioxide that occur due to changes in tree structure and density in montane Norway spruce stands of Central Germany were analyzed with a three dimensional microclimate and gas exchange model STANDFLUX. The model was used to calculate forest radiation absorption, the net photosynthesis and transpiration of single trees, and gas exchange of tree canopies. Model parameterizations were derived for six stands of Picea abies ...

  17. Origin and speciation of Picea schrenkiana and Piceasmithiana in the Center Asian Highlands and Himalayas.

    Science.gov (United States)

    Li, Lili; Sun, Yongshuai; Zou, Jiabin; Yue, Wei; Wang, Xi; Liu, Jianquan

    Elucidating the evolutionary history of current species diversity, especially trees with large effective population sizes and long generation times, is a complicated exercise confounded by gene flow and incomplete lineage sorting. In the present study, we aim to determine the origin and speciation of Picea schrenkiana and Picea smithiana using population genetic data from chloroplast (cp), mitochondrial (mt), and nuclear (nr) genomes. These two species occur in the Central Asian Highlands and Himalayas, respectively, where they are isolated from other Asian congeneric species by the Qinghai-Tibet Plateau (QTP) or adjacent deserts. Previous studies based on both morphological and molecular evidence suggest that they have contrasting phylogenetic relationships with Picea likiangensis or Picea wilsonii which are closely related and both located in the QTP. We examined genetic variation among 16 loci of three genomes from 30 populations of these four species. At both cpDNA loci and mtDNA loci, P. schrenkiana appeared to be closely related to P. likiangensis , although statistical support for this was weak. However, phylogenetic analyses and speciation tests based on the nuclear data from 11 loci provided evidence that P. schrenkiana and P. smithiana are sister species. These two species diverged around five million years ago (Mya) while the divergence between them and the P. likiangensis - P. wilsonii clade occurred about 18.4 Mya. We also detected gene flow accompanying these speciation events. Our results highlight the complex speciation histories of these alpine conifers due to interspecific gene flow and/or incomplete lineage sorting, and the importance of the early QTP uplifts in promoting the origin of these important conifer species in the Asian highlands.

  18. Effects of deicing salt on the vitality and health of two spruce species, Picea abies Karst., and Picea glehnii Masters planted along roadsides in northern Japan

    International Nuclear Information System (INIS)

    Kayama, M.; Quoreshi, A.M.; Kitaoka, S.; Kitahashi, Y.; Sakamoto, Y.; Maruyama, Y.; Kitao, M.; Koike, T.

    2003-01-01

    Innate physiological characters of conifers may increase uptake of sodium and chloride and result in enhanced tree injury. - In northern Japan, the growth of Picea abies Karst., and Picea glehnii Masters, which have been planted along the highways, is often suppressed due to several environmental stresses. To examine the adverse effects of deicing salt, the primary source of stress, we measured needle life span, photosynthetic capacity, and water potential and transpiration rate of the two spruce species at a site with damaged trees, near the roadside and a site with healthy trees, located far from the highway. Results from the analysis showed large amounts of sodium and chlorine in the soil and snow at the damaged site. These elements had accumulated in the needles of the spruce. Moreover, physiological traits of the spruce, at the damaged site were also affected. Therefore, we concluded that poor physiological traits might be attributed to an accumulation of deicing salt in the needles, resulting in the suppression of tree growth

  19. Bioreactor scale up and protein product quality characterization of piggyBac transposon derived CHO pools.

    Science.gov (United States)

    Rajendra, Yashas; Balasubramanian, Sowmya; Peery, Robert B; Swartling, James R; McCracken, Neil A; Norris, Dawn L; Frye, Christopher C; Barnard, Gavin C

    2017-03-01

    Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies, and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development. We recently described the generation of CHO pools yielding mAb titers as high as 7.6 g/L in a 16 day bioprocess using piggyBac transposon-mediated gene integration. In this study, we wanted to understand why the piggyBac pool titers were significantly higher (2-10 fold) than the control CHO pools. Higher titers were the result of a combination of increased average gene copy number, significantly higher messenger RNA levels and the homogeneity (i.e. less diverse population distribution) of the piggyBac pools, relative to the control pools. In order to validate the use of piggyBac pools to support preclinical drug development, we then performed an in-depth product quality analysis of purified protein. The product quality of protein obtained from the piggyBac pools was very similar to the product quality profile of protein obtained from the control pools. Finally, we demonstrated the scalability of these pools from shake flasks to 36L bioreactors. Overall, these results suggest that gram quantities of therapeutic protein can be rapidly obtained from piggyBac CHO pools without significantly changing product quality attributes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:534-540, 2017. © 2017 American Institute of Chemical Engineers.

  20. The complexity of Rhipicephalus (Boophilus microplus genome characterised through detailed analysis of two BAC clones

    Directory of Open Access Journals (Sweden)

    Valle Manuel

    2011-07-01

    Full Text Available Abstract Background Rhipicephalus (Boophilus microplus (Rmi a major cattle ectoparasite and tick borne disease vector, impacts on animal welfare and industry productivity. In arthropod research there is an absence of a complete Chelicerate genome, which includes ticks, mites, spiders, scorpions and crustaceans. Model arthropod genomes such as Drosophila and Anopheles are too taxonomically distant for a reference in tick genomic sequence analysis. This study focuses on the de-novo assembly of two R. microplus BAC sequences from the understudied R microplus genome. Based on available R. microplus sequenced resources and comparative analysis, tick genomic structure and functional predictions identify complex gene structures and genomic targets expressed during tick-cattle interaction. Results In our BAC analyses we have assembled, using the correct positioning of BAC end sequences and transcript sequences, two challenging genomic regions. Cot DNA fractions compared to the BAC sequences confirmed a highly repetitive BAC sequence BM-012-E08 and a low repetitive BAC sequence BM-005-G14 which was gene rich and contained short interspersed elements (SINEs. Based directly on the BAC and Cot data comparisons, the genome wide frequency of the SINE Ruka element was estimated. Using a conservative approach to the assembly of the highly repetitive BM-012-E08, the sequence was de-convoluted into three repeat units, each unit containing an 18S, 5.8S and 28S ribosomal RNA (rRNA encoding gene sequence (rDNA, related internal transcribed spacer and complex intergenic region. In the low repetitive BM-005-G14, a novel gene complex was found between to 2 genes on the same strand. Nested in the second intron of a large 9 Kb papilin gene was a helicase gene. This helicase overlapped in two exonic regions with the papilin. Both these genes were shown expressed in different tick life stage important in ectoparasite interaction with the host. Tick specific sequence

  1. Comparison of biological activated carbon (BAC) and membrane bioreactor (MBR) for pollutants removal in drinking water treatment.

    Science.gov (United States)

    Tian, J Y; Chen, Z L; Liang, H; Li, X; Wang, Z Z; Li, G B

    2009-01-01

    Biological activated carbon (BAC) and membrane bioreactor (MBR) were systematically compared for the drinking water treatment from slightly polluted raw water under the same hydraulic retention time (HRT) of 0.5 h. MBR exhibited excellent turbidity removal capacity due to the separation of the membrane; while only 60% of influent turbidity was intercepted by BAC. Perfect nitrification was achieved by MBR with the 89% reduction in ammonia; by contrast, BAC only eliminated a moderate amount of influent ammonia (by 54.5%). However, BAC was able to remove more dissolved organic matter (DOM, especially for organic molecules of 3,000 approximately 500 Daltons) and corresponding disinfection by-product formation potential (DBPFP) in raw water than MBR. Unfortunately, particulate organic matter (POM) was detected in the BAC effluent. On the other hand, BAC and MBR displayed essentially the same capacity for biodegradable organic matter (BOM) removal. Fractionation of DOM showed that the removal efficiencies of hydrophobic neutrals, hydrophobic acids, weakly hydrophobic acids and hydrophilic organic matter through BAC treatment were 11.7%, 8.8%, 13.9% and 4.8% higher than that through MBR; while MBR achieved 13.8% higher hydrophobic bases removal as compared with BAC.

  2. State Blood Alcohol Concentration (BAC) Testing and Reporting for Drivers Involved in Fatal Crashes : Current Practices, Results, and Strategies, 1997-2009

    Science.gov (United States)

    2012-08-01

    This report documents current State blood alcohol concentration (BAC) testing and reporting practices and results for drivers involved in fatal crashes. It summarizes known BAC results by State for the years 1997 to 2009 for both fatally injured and ...

  3. Interspecific transfer of only part of genome by fusion between non-irradiated protoplasts of Nicotiana glauca and X-ray irradiated protoplasts of N. Langsdorffii

    International Nuclear Information System (INIS)

    Itoh, K.; Futsuhara, Y.

    1983-01-01

    To transfer only part of genome, X-ray irradiated suspension cell protoplasts of N. langsdorffii were fused with suspension cell protoplasts of N. glauca by polyethylene glycol. Somatic hybrid calli were selected by the growth in the hormone-free medium. Some of somatic hybrid calli from fusion with irradiated protoplasts indicated the loss of small subunit polypeptide of fraction 1 protein which was coded by N. langsdorffii nuclear DNA. Cytological analysis provided an information on significant decrease of chromosomes in somatic hybrid calli from fusion with irradiated protoplasts, compared with the somatic hybrid calli from fusion with non-irradiated protoplasts. In addition, isozyme analysis revealed that somatic hybrid calli from fusion with irradiated protoplasts lost particular bands of N. langsdorffli. These results demonstrate the tranfer of only part of genome from N, langsdorffii to N, glauca by fusion with X-ray irradiated protoplasts

  4. The bacterial artificial chromosome (BAC) library of the narrow-leafed lupin (Lupinus angustifolius L.)

    Czech Academy of Sciences Publication Activity Database

    Kasprzak, A.; Šafář, Jan; Janda, Jaroslav; Doležel, Jaroslav; Wolko, B.; Naganowska, B.

    2006-01-01

    Roč. 11, - (2006), s. 396-407 ISSN 1425-8153 R&D Projects: GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : BAC * genomic DNA library * Lupinus angustifolius L. Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.238, year: 2006

  5. A PiggyBac-mediated approach for muscle gene transfer or cell therapy

    Directory of Open Access Journals (Sweden)

    Déborah Ley

    2014-11-01

    Full Text Available An emerging therapeutic approach for Duchenne muscular dystrophy is the transplantation of autologous myogenic progenitor cells genetically modified to express dystrophin. The use of this approach is challenged by the difficulty in maintaining these cells ex vivo while keeping their myogenic potential, and ensuring sufficient transgene expression following their transplantation and myogenic differentiation in vivo. We investigated the use of the piggyBac transposon system to achieve stable gene expression when transferred to cultured mesoangioblasts and into murine muscles. Without selection, up to 8% of the mesoangioblasts expressed the transgene from 1 to 2 genomic copies of the piggyBac vector. Integration occurred mostly in intergenic genomic DNA and transgene expression was stable in vitro. Intramuscular transplantation of mouse Tibialis anterior muscles with mesoangioblasts containing the transposon led to sustained myofiber GFP expression in vivo. In contrast, the direct electroporation of the transposon-donor plasmids in the mouse Tibialis muscles in vivo did not lead to sustained transgene expression despite molecular evidence of piggyBac transposition in vivo. Together these findings provide a proof-of-principle that piggyBac transposon may be considered for mesoangioblast cell-based therapies of muscular dystrophies.

  6. Insect transformation with piggyBac: getting the number of injections just right

    Science.gov (United States)

    Morrison, N. I.; Shimeld, S. M.

    2016-01-01

    Abstract The insertion of exogenous genetic cargo into insects using transposable elements is a powerful research tool with potential applications in meeting food security and public health challenges facing humanity. piggyBac is the transposable element most commonly utilized for insect germline transformation. The described efficiency of this process is variable in the published literature, and a comprehensive review of transformation efficiency in insects is lacking. This study compared and contrasted all available published data with a comprehensive data set provided by a biotechnology group specializing in insect transformation. Based on analysis of these data, with particular focus on the more complete observational data from the biotechnology group, we designed a decision tool to aid researchers' decision‐making when using piggyBac to transform insects by microinjection. A combination of statistical techniques was used to define appropriate summary statistics of piggyBac transformation efficiency by species and insect order. Publication bias was assessed by comparing the data sets. The bias was assessed using strategies co‐opted from the medical literature. The work culminated in building the Goldilocks decision tool, a Markov‐Chain Monte‐Carlo simulation operated via a graphical interface and providing guidance on best practice for those seeking to transform insects using piggyBac. PMID:27027400

  7. Differential CT features of infectious pneumonia versus bronchioloalveolar carcinoma (BAC) mimicking pneumonia

    International Nuclear Information System (INIS)

    Kim, Tae Hoon; Kim, Sang Jin; Ryu, Young Hoon; Chung, Soo Yoon; Seo, Jae Seung; Kim, Young Jin; Choi, Byoung Wook; Lee, Sun Hwa; Cho, Sang Ho

    2006-01-01

    The purpose of this study was to evaluate retrospectively the differential CT features of bronchioloalveolar carcinoma (BAC) mimicking pneumonia and infectious pneumonia at the lung periphery. CT images were reviewed in 47 patients with focal areas of parenchymal opacification at the lung periphery. We evaluated the presence of ground-glass attenuation, marginal conspicuity of the lesion, CT angiogram sign, air-bronchogram sign, a bubble-like low-attenuation area within the lesion, presence of pleural thickening and retraction associated with the lesion, presence of pleural effusion and extra-pleural fatty hypertrophy, presence of bronchial wall thickening proximal to the lesion, and air-trapping in the normal lung near the lesion. BAC (n=18) depicted the presence of a bubble-like low-attenuation area within the lesion, whereas infectious pneumonia (n=29) represented the pleural thickening associated with the lesion and bronchial wall thickening proximal to the lesion (P 0.05). The focal areas of the parenchymal opacification on the CT images may suggest infectious pneumonia rather than BAC when they show bronchial wall thickening proximal to the lesion and pleural thickening associated with the lesion, whereas BAC is characterized as the presence of a bubble-like low attenuation area within the tumor. (orig.)

  8. Measuring brain activity cycling (BAC) in long term EEG monitoring of preterm babies

    International Nuclear Information System (INIS)

    Stevenson, Nathan J; Palmu, Kirsi; Wikström, Sverre; Hellström-Westas, Lena; Vanhatalo, Sampsa

    2014-01-01

    Measuring fluctuation of vigilance states in early preterm infants undergoing long term intensive care holds promise for monitoring their neurological well-being. There is currently, however, neither objective nor quantitative methods available for this purpose in a research or clinical environment. The aim of this proof-of-concept study was, therefore, to develop quantitative measures of the fluctuation in vigilance states or brain activity cycling (BAC) in early preterm infants. The proposed measures of BAC were summary statistics computed on a frequency domain representation of the proportional duration of spontaneous activity transients (SAT%) calculated from electroencephalograph (EEG) recordings. Eighteen combinations of three statistics and six frequency domain representations were compared to a visual interpretation of cycling in the SAT% signal. Three high performing measures (band energy/periodogram: R = 0.809, relative band energy/nonstationary frequency marginal: R = 0.711, g-statistic/nonstationary frequency marginal: R = 0.638) were then compared to a grading of sleep wake cycling based on the visual interpretation of the amplitude-integrated EEG trend. These measures of BAC are conceptually straightforward, correlate well with the visual scores of BAC and sleep wake cycling, are robust enough to cope with the technically compromised monitoring data available in intensive care units, and are recommended for further validation in prospective studies. (paper)

  9. BacHBerry:: BACterial Hosts for production of Bioactive phenolics from bERRY fruits

    DEFF Research Database (Denmark)

    Dudnik, Alexey; Almeida, A. Filipa; Andrade, Ricardo

    2017-01-01

    BACterial Hosts for production of Bioactive phenolics from bERRY fruits (BacHBerry) was a 3-year project funded by the Seventh Framework Programme (FP7) of the European Union that ran between November 2013 and October 2016. The overall aim of the project was to establish a sustainable and economi...

  10. Isolation of Specific Clones from Nonarrayed BAC Libraries through Homologous Recombination

    Directory of Open Access Journals (Sweden)

    Mikhail Nefedov

    2011-01-01

    Full Text Available We have developed a new approach to screen bacterial artificial chromosome (BAC libraries by recombination selection. To test this method, we constructed an orangutan BAC library using an E. coli strain (DY380 with temperature inducible homologous recombination (HR capability. We amplified one library segment, induced HR at 42∘C to make it recombination proficient, and prepared electrocompetent cells for transformation with a kanamycin cassette to target sequences in the orangutan genome through terminal recombineering homologies. Kanamycin-resistant colonies were tested for the presence of BACs containing the targeted genes by the use of a PCR-assay to confirm the presence of the kanamycin insertion. The results indicate that this is an effective approach for screening clones. The advantage of recombination screening is that it avoids the high costs associated with the preparation, screening, and archival storage of arrayed BAC libraries. In addition, the screening can be conceivably combined with genetic engineering to create knockout and reporter constructs for functional studies.

  11. Differential CT features of infectious pneumonia versus bronchioloalveolar carcinoma (BAC) mimicking pneumonia

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Tae Hoon [Yonsei University College of Medicine, Department of Radiology, Seoul (Korea); Yongdong Severance Hospital, Department of Radiology, Seoul (Korea); Kim, Sang Jin; Ryu, Young Hoon; Chung, Soo Yoon; Seo, Jae Seung; Kim, Young Jin; Choi, Byoung Wook [Yonsei University College of Medicine, Department of Radiology, Seoul (Korea); Lee, Sun Hwa [NeoDin Medical Institute, Department of Clinical Pathology, Seoul (Korea); Cho, Sang Ho [Yonsei University College of Medicine, Department of Pathology, Seoul (Korea)

    2006-08-15

    The purpose of this study was to evaluate retrospectively the differential CT features of bronchioloalveolar carcinoma (BAC) mimicking pneumonia and infectious pneumonia at the lung periphery. CT images were reviewed in 47 patients with focal areas of parenchymal opacification at the lung periphery. We evaluated the presence of ground-glass attenuation, marginal conspicuity of the lesion, CT angiogram sign, air-bronchogram sign, a bubble-like low-attenuation area within the lesion, presence of pleural thickening and retraction associated with the lesion, presence of pleural effusion and extra-pleural fatty hypertrophy, presence of bronchial wall thickening proximal to the lesion, and air-trapping in the normal lung near the lesion. BAC (n=18) depicted the presence of a bubble-like low-attenuation area within the lesion, whereas infectious pneumonia (n=29) represented the pleural thickening associated with the lesion and bronchial wall thickening proximal to the lesion (P<0.05). The other CT findings showed no significant differences (P>0.05). The focal areas of the parenchymal opacification on the CT images may suggest infectious pneumonia rather than BAC when they show bronchial wall thickening proximal to the lesion and pleural thickening associated with the lesion, whereas BAC is characterized as the presence of a bubble-like low attenuation area within the tumor. (orig.)

  12. BAC and Beer: Operationalizing Drunk Driving Laws in a Research Methods Course Exercise.

    Science.gov (United States)

    Taylor, Ralph B.; McConnell, Patrick

    2001-01-01

    Focuses on an exercise utilized in a research methods class and based on social problems that invites student interest. Explains the exercise has students determine their blood alcohol level (BAC) by asking them to estimate the number of beers it would take to have them just reach driving under the influence (DUI) status. (CMK)

  13. Advancing Eucalyptus genomics: identification and sequencing of lignin biosynthesis genes from deep-coverage BAC libraries

    Directory of Open Access Journals (Sweden)

    Kudrna David

    2011-03-01

    Full Text Available Abstract Background Eucalyptus species are among the most planted hardwoods in the world because of their rapid growth, adaptability and valuable wood properties. The development and integration of genomic resources into breeding practice will be increasingly important in the decades to come. Bacterial artificial chromosome (BAC libraries are key genomic tools that enable positional cloning of important traits, synteny evaluation, and the development of genome framework physical maps for genetic linkage and genome sequencing. Results We describe the construction and characterization of two deep-coverage BAC libraries EG_Ba and EG_Bb obtained from nuclear DNA fragments of E. grandis (clone BRASUZ1 digested with HindIII and BstYI, respectively. Genome coverages of 17 and 15 haploid genome equivalents were estimated for EG_Ba and EG_Bb, respectively. Both libraries contained large inserts, with average sizes ranging from 135 Kb (Eg_Bb to 157 Kb (Eg_Ba, very low extra-nuclear genome contamination providing a probability of finding a single copy gene ≥ 99.99%. Libraries were screened for the presence of several genes of interest via hybridizations to high-density BAC filters followed by PCR validation. Five selected BAC clones were sequenced and assembled using the Roche GS FLX technology providing the whole sequence of the E. grandis chloroplast genome, and complete genomic sequences of important lignin biosynthesis genes. Conclusions The two E. grandis BAC libraries described in this study represent an important milestone for the advancement of Eucalyptus genomics and forest tree research. These BAC resources have a highly redundant genome coverage (> 15×, contain large average inserts and have a very low percentage of clones with organellar DNA or empty vectors. These publicly available BAC libraries are thus suitable for a broad range of applications in genetic and genomic research in Eucalyptus and possibly in related species of Myrtaceae

  14. Genetic diversity in populations of Isatis glauca Aucher ex Boiss. ssp. from Central Anatolia in Turkey, as revealed by AFLP analysis.

    Science.gov (United States)

    Özbek, Özlem; Görgülü, Elçin; Yıldırımlı, Şinasi

    2013-12-01

    Isatidae L. is a complex and systematically difficult genus in Brassicaceae. The genus displays great morphological polymorphism, which makes the classification of species and subspecies difficult as it is observed in Isatis glauca Aucher ex Boiss. The aim of this study is characterization of the genetic diversity in subspecies of Isatis glauca Aucher ex Boiss. distributed widely in Central Anatolia, in Turkey by using Amplified Fragment Length Polymorphism (AFLP) technique. Eight different Eco RI-Mse I primer combinations produced 805 AFLP loci, 793 (98.5%) of which were polymorphic in 67 accessions representing nine different populations. The data obtained by AFLP was computed with using GDA (Genetic Data Analysis) and STRUCTURE (version 2.3.3) software programs for population genetics. The mean proportion of the polymorphic locus (P), the mean number of alleles (A), the number of unique alleles (U) and the mean value of gene diversity (He) were 0.59, 1.59, 20, and 0.23 respectively. The coancestry coefficient (ϴ) was 0.24. The optimal number of K was identified as seven. The principal component analysis (PCA) explained 85.61% of the total genetic variation. Isatis glauca ssp. populations showed a high level of genetic diversity, and the AFLP analysis revealed that high polymorphism and differentiated subspecies could be used conveniently for population genetic studies. The principal coordinate analysis (PCoA) based on the dissimilarity matrix, the dendrogram drawn with UPGMA method and STRUCTURE cluster analysis distinguished the accessions successfully. The accessions formed distinctive population structures for populations AA, AB, E, K, and S. Populations AG1 and AG2 seemed to have similar genetic content, in addition, in both populations several hybrid individuals were observed. The accessions did not formed distinctive population structures for both populations AI and ANP. Consequently, Ankara province might be the area, where species Isatis glauca Aucher

  15. Comparative analysis of catfish BAC end sequences with the zebrafish genome

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    Abernathy Jason

    2009-12-01

    Full Text Available Abstract Background Comparative mapping is a powerful tool to transfer genomic information from sequenced genomes to closely related species for which whole genome sequence data are not yet available. However, such an approach is still very limited in catfish, the most important aquaculture species in the United States. This project was initiated to generate additional BAC end sequences and demonstrate their applications in comparative mapping in catfish. Results We reported the generation of 43,000 BAC end sequences and their applications for comparative genome analysis in catfish. Using these and the additional 20,000 existing BAC end sequences as a resource along with linkage mapping and existing physical map, conserved syntenic regions were identified between the catfish and zebrafish genomes. A total of 10,943 catfish BAC end sequences (17.3% had significant BLAST hits to the zebrafish genome (cutoff value ≤ e-5, of which 3,221 were unique gene hits, providing a platform for comparative mapping based on locations of these genes in catfish and zebrafish. Genetic linkage mapping of microsatellites associated with contigs allowed identification of large conserved genomic segments and construction of super scaffolds. Conclusion BAC end sequences and their associated polymorphic markers are great resources for comparative genome analysis in catfish. Highly conserved chromosomal regions were identified to exist between catfish and zebrafish. However, it appears that the level of conservation at local genomic regions are high while a high level of chromosomal shuffling and rearrangements exist between catfish and zebrafish genomes. Orthologous regions established through comparative analysis should facilitate both structural and functional genome analysis in catfish.

  16. Membrane Topology and Biochemical Characterization of the Escherichia coli BacA Undecaprenyl-Pyrophosphate Phosphatase.

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    Guillaume Manat

    Full Text Available Several integral membrane proteins exhibiting undecaprenyl-pyrophosphate (C55-PP phosphatase activity were previously identified in Escherichia coli that belonged to two distinct protein families: the BacA protein, which accounts for 75% of the C55-PP phosphatase activity detected in E. coli cell membranes, and three members of the PAP2 phosphatidic acid phosphatase family, namely PgpB, YbjG and LpxT. This dephosphorylation step is required to provide the C55-P carrier lipid which plays a central role in the biosynthesis of various cell wall polymers. We here report detailed investigations of the biochemical properties and membrane topology of the BacA protein. Optimal activity conditions were determined and a narrow-range substrate specificity with a clear preference for C55-PP was observed for this enzyme. Alignments of BacA protein sequences revealed two particularly well-conserved regions and several invariant residues whose role in enzyme activity was questioned by using a site-directed mutagenesis approach and complementary in vitro and in vivo activity assays. Three essential residues Glu21, Ser27, and Arg174 were identified, allowing us to propose a catalytic mechanism for this enzyme. The membrane topology of the BacA protein determined here experimentally did not validate previous program-based predicted models. It comprises seven transmembrane segments and contains in particular two large periplasmic loops carrying the highly-conserved active site residues. Our data thus provide evidence that all the different E. coli C55-PP phosphatases identified to date (BacA and PAP2 catalyze the dephosphorylation of C55-PP molecules on the same (outer side of the plasma membrane.

  17. A BAC/BIBAC-based physical map of chickpea, Cicer arietinum L

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    Abbo Shahal

    2010-09-01

    Full Text Available Abstract Background Chickpea (Cicer arietinum L. is the third most important pulse crop worldwide. Despite its importance, relatively little is known about its genome. The availability of a genome-wide physical map allows rapid fine mapping of QTL, development of high-density genome maps, and sequencing of the entire genome. However, no such a physical map has been developed in chickpea. Results We present a genome-wide, BAC/BIBAC-based physical map of chickpea developed by fingerprint analysis. Four chickpea BAC and BIBAC libraries, two of which were constructed in this study, were used. A total of 67,584 clones were fingerprinted, and 64,211 (~11.7 × of the fingerprints validated and used in the physical map assembly. The physical map consists of 1,945 BAC/BIBAC contigs, with each containing an average of 28.3 clones and having an average physical length of 559 kb. The contigs collectively span approximately 1,088 Mb. By using the physical map, we identified the BAC/BIBAC contigs containing or closely linked to QTL4.1 for resistance to Didymella rabiei (RDR and QTL8 for days to first flower (DTF, thus further verifying the physical map and confirming its utility in fine mapping and cloning of QTL. Conclusion The physical map represents the first genome-wide, BAC/BIBAC-based physical map of chickpea. This map, along with other genomic resources previously developed in the species and the genome sequences of related species (soybean, Medicago and Lotus, will provide a foundation necessary for many areas of advanced genomics research in chickpea and other legume species. The inclusion of transformation-ready BIBACs in the map greatly facilitates its utility in functional analysis of the legume genomes.

  18. 454 sequencing of pooled BAC clones on chromosome 3H of barley

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    Yamaji Nami

    2011-05-01

    Full Text Available Abstract Background Genome sequencing of barley has been delayed due to its large genome size (ca. 5,000Mbp. Among the fast sequencing systems, 454 liquid phase pyrosequencing provides the longest reads and is the most promising method for BAC clones. Here we report the results of pooled sequencing of BAC clones selected with ESTs genetically mapped to chromosome 3H. Results We sequenced pooled barley BAC clones using a 454 parallel genome sequencer. A PCR screening system based on primer sets derived from genetically mapped ESTs on chromosome 3H was used for clone selection in a BAC library developed from cultivar "Haruna Nijo". The DNA samples of 10 or 20 BAC clones were pooled and used for shotgun library development. The homology between contig sequences generated in each pooled library and mapped EST sequences was studied. The number of contigs assigned on chromosome 3H was 372. Their lengths ranged from 1,230 bp to 58,322 bp with an average 14,891 bp. Of these contigs, 240 showed homology and colinearity with the genome sequence of rice chromosome 1. A contig annotation browser supplemented with query search by unique sequence or genetic map position was developed. The identified contigs can be annotated with barley cDNAs and reference sequences on the browser. Homology analysis of these contigs with rice genes indicated that 1,239 rice genes can be assigned to barley contigs by the simple comparison of sequence lengths in both species. Of these genes, 492 are assigned to rice chromosome 1. Conclusions We demonstrate the efficiency of sequencing gene rich regions from barley chromosome 3H, with special reference to syntenic relationships with rice chromosome 1.

  19. Construction of a BAC library and identification of Dmrt1 gene of the rice field eel, Monopterus albus

    International Nuclear Information System (INIS)

    Jang Songhun; Zhou Fang; Xia Laixin; Zhao Wei; Cheng Hanhua; Zhou Rongjia

    2006-01-01

    A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA from the rice field eel (Monopterus albus). The BAC library consists of a total of 33,000 clones with an average insert size of 115 kb. Based on the rice field eel haploid genome size of 600 Mb, the BAC library is estimated to contain approximately 6.3 genome equivalents and represents 99.8% of the genome of the rice field eel. This is first BAC library constructed from this species. To estimate the possibility of isolating a specific clone, high-density colony hybridization-based library screening was performed using Dmrt1 cDNA of the rice field eel as a probe. Both library screening and PCR identification results revealed three positive BAC clones which were overlapped, and formed a contig covering the Dmrt1 gene of 195 kb. By sequence comparisons with the Dmrt1 cDNA and sequencing of first four intron-exon junctions, Dmrt1 gene of the rice field eel was predicted to contain four introns and five exons. The sizes of first and second intron are 1.5 and 2.6 kb, respectively, and the sizes of last two introns were predicted to be about 20 kb. The Dmrt1 gene structure was conserved in evolution. These results also indicate that the BAC library is a useful resource for BAC contig construction and molecular isolation of functional genes

  20. Early testing of adaptedness to temperature and water availability in Pinus sylvestris and Picea abies

    International Nuclear Information System (INIS)

    Sonesson, Johan

    2000-01-01

    Long-term climate changes have been evident in the past. In the future an increase in the rate of climate change is predicted owing to man-made emissions. Studies of adaptedness to different climatic conditions are of great importance for the design of appropriate breeding and gene conservation programmes. This thesis presents studies of adaptedness to temperature and water availability in Pinus sylvestris and Picea abies and explores the possibilities of utilising differences in adaptedness to obtain juvenile-mature (J-M) correlations strong enough for efficient early testing. Offspring from clones in two Swedish Pinus sylvestris seed orchards and one Picea abies seed orchard were grown in growth chambers for one and two growth periods respectively. Two temperature regimes and two irrigation regimes were applied in a factorial design. Both species expressed high phenotypic plasticity and additive variance for height growth and biomass traits. This implies that these populations should be able to adapt both to short-term and to long-term climate changes. Genotype by environment (GxE) interaction indicated strong differences in adaptedness to temperature and low differences in adaptedness to water availability. Parent rank changes between treatments indicated that climate change could seriously alter the ranking of clones in breeding populations and thus decrease the genetic gain obtained in previous selections. Differences in stability among parents suggested that culling of unstable genotypes could be a way to reduce the negative effects of GxE interaction. Genetic correlations between growth chamber and 14-30 year old field progeny trials with the same parents were mainly weak for both species. The correlations were improved by the drought treatment in the Picea abies experiment suggesting that further development of early testing methods for this species should include treatments with limiting water availability

  1. The first insight into the salvia (lamiaceae) genome via bac library construction and high-throughput sequencing of target bac clones

    International Nuclear Information System (INIS)

    Hao, D.C.; Vautrin, S.; Berges, H.; Chen, S.L.

    2015-01-01

    Salvia is a representative genus of Lamiaceae, a eudicot family with significant species diversity and population adaptibility. One of the key goals of Salvia genomics research is to identify genes of adaptive significance. This information may help to improve the conservation of adaptive genetic variation and the management of medicinal plants to increase their health and productivity. Large-insert genomic libraries are a fundamental tool for achieving this purpose. We report herein the construction, characterization and screening of a gridded BAC library for Salvia officinalis (sage). The S. officinalis BAC library consists of 17,764 clones and the average insert size is 107 Kb, corresponding to 3 haploid genome equivalents. Seventeen positive clones (average insert size 115 Kb) containing five terpene synthase (TPS) genes were screened out by PCR and 12 of them were subject to Illumina HiSeq 2000 sequencing, which yielded 28,097,480 90-bp raw reads (2.53 Gb). Scaffolds containing sabinene synthase (Sab), a Sab homolog, TPS3 (kaurene synthase-like 2), copalyl diphosphate synthase 2 and one cytochrome P450 gene were retrieved via de novo assembly and annotation, which also have flanking noncoding sequences, including predicted promoters and repeat sequences. Among 2,638 repeat sequences, there are 330 amplifiable microsatellites. This BAC library provides a new resource for Lamiaceae genomic studies, including microsatellite marker development, physical mapping, comparative genomics and genome sequencing. Characterization of positive clones provided insights into the structure of the Salvia genome. These sequences will be used in the assembly of a future genome sequence for S. officinalis. (author)

  2. Investigation of decolorization of textile wastewater in an anaerobic/aerobic biological activated carbon system (A/A BAC).

    Science.gov (United States)

    Pasukphun, N; Vinitnantharat, S; Gheewala, S

    2010-04-01

    The aim of this study is to investigate the decolorization in anaerobic/aerobic biological activated carbon (A/A BAC) system. The experiment was divided into 2 stages; stage I is batch test for preliminary study of dye removal equilibrium time. The preliminary experiment (stage I) provided the optimal data for experimental design of A/A BAC system in SBR (stage II). Stage II is A/A BAC system imitated Sequencing Batch Reactor (SBR) which consist of 5 main periods; fill, react, settle, draw and idle. React period include anaerobic phase followed by aerobic phase. The BAC main media; Granular Activated Carbon (GAC), Mixed Cultures (MC) and Biological Activated Carbon (BAC) were used for dye and organic substances removal in three different solutions; Desizing Agent Solution (DAS), dye Solution (DS) and Synthetic Textile Wastewater (STW). Results indicate that GAC adsorption plays role in dye removal followed by BAC and MC activities, respectively. In the presence desizing agent, decolorization by MC was improved because desizing agent acts as co-substrates for microorganisms. It was found that 50% of dye removal efficiency was achieved in Fill period by MC. GC/MS analysis was used to identify dye intermediate from decolorization. Dye intermediate containing amine group was found in the solution and on BAC surfaces. The results demonstrated that combination of MC and BAC in the system promotes decolorization and dye intermediate removal. In order to improve dye removal efficiency in an A/A BAC system, replacement of virgin GAC, sufficient co-substrates supply and the appropriate anaerobic: aerobic period should be considered.

  3. Potential therapeutic agents for circulatory diseases from Bauhinia glauca Benth.subsp. pernervosa. (Da Ye Guan Men).

    Science.gov (United States)

    Tang, Yingzhan; Ling, Junhong; Zhang, Peng; Zhang, Xiangrong; Zhang, Na; Wang, Wenli; Li, Jiayuan; Li, Ning

    2015-08-15

    Because of platelets as critical factor in the formation of pathogenic thrombi, anti-platelet activities have been selected as therapeutic target for various circulatory diseases. In order to find potential therapeutic agents, bioassay-directed separation of Bauhinia glauca Benth.subsp. pernervosa. (called Da Ye Guan Men as a traditional Chinese medicine) was performed to get 29 main components (compounds 1-29) from the bioactive part of this herbal. It was the first time to focus on the composition with anti-platelet aggregation activities for this traditional Chinese medicine. The constituents, characterized from the effective extract, were established on the basis of extensive spectral data analysis. Then their anti-platelet aggregation effects were evaluated systematically. On the basis of the chemical profile and biological assay, it was suggested that the flavonoid composition (5 and 18) should be responsible for the anti-platelet aggregation of the herbal because of their significant activities. The primary structure and activity relationship was also discussed briefly. Copyright © 2015. Published by Elsevier Ltd.

  4. World without borders-genetic population structure of a highly migratory marine predator, the blue shark (Prionace glauca).

    Science.gov (United States)

    Veríssimo, Ana; Sampaio, Íris; McDowell, Jan R; Alexandrino, Paulo; Mucientes, Gonzalo; Queiroz, Nuno; da Silva, Charlene; Jones, Catherine S; Noble, Leslie R

    2017-07-01

    Highly migratory, cosmopolitan oceanic sharks often exhibit complex movement patterns influenced by ontogeny, reproduction, and feeding. These elusive species are particularly challenging to population genetic studies, as representative samples suitable for inferring genetic structure are difficult to obtain. Our study provides insights into the genetic population structure one of the most abundant and wide-ranging oceanic shark species, the blue shark Prionace glauca, by sampling the least mobile component of the populations, i.e., young-of-year and small juveniles (<2 year; N  = 348 individuals), at three reported nursery areas, namely, western Iberia, Azores, and South Africa. Samples were collected in two different time periods (2002-2008 and 2012-2015) and were screened at 12 nuclear microsatellites and at a 899-bp fragment of the mitochondrial control region. Our results show temporally stable genetic homogeneity among the three Atlantic nurseries at both nuclear and mitochondrial markers, suggesting basin-wide panmixia. In addition, comparison of mtDNA CR sequences from Atlantic and Indo-Pacific locations also indicated genetic homogeneity and unrestricted female-mediated gene flow between ocean basins. These results are discussed in light of the species' life history and ecology, but suggest that blue shark populations may be connected by gene flow at the global scale. The implications of the present findings to the management of this important fisheries resource are also discussed.

  5. Immediate effects of nectar robbing by Palestine sunbirds (Nectarinia osea) on nectar alkaloid concentrations in tree tobacco (Nicotiana glauca).

    Science.gov (United States)

    Kaczorowski, Rainee L; Koplovich, Avi; Sporer, Frank; Wink, Michael; Markman, Shai

    2014-04-01

    Plant secondary metabolites (PSMs), such as alkaloids, are often found in many parts of a plant, including flowers, providing protection to the plant from various types of herbivores or microbes. PSMs are also present in the floral nectar of many species, but typically at lower concentrations than in other parts of the plant. Nectar robbers often damage floral tissue to access the nectar. By doing so, these nectar robbers may initiate an increase of PSMs in the floral nectar. It is often assumed that it takes at least a few hours before the plant demonstrates an increase in PSMs. Here, we addressed the question of whether PSMs in the floral tissue are immediately being released into the floral nectar following nectar robbing. To address this research question, we investigated whether there was an immediate effect of nectar robbing by the Palestine Sunbird (Nectarinia osea) on the concentration of nectar alkaloids, nicotine and anabasine, in Tree Tobacco (Nicotiana glauca). We found that the concentration of anabasine, but not nicotine, significantly increased in floral nectar immediately following simulated nectar robbing. These findings suggest that nectar robbers could be ingesting greater amounts of PSMs than they would if they visit flowers legitimately. As a consequence, increased consumption of neurotoxic nectar alkaloids or other PSMs could have negative effects on the nectar robber.

  6. Morphological analysis of the oviduct, oviducal gland and isthmus of the blue shark Prionace glauca (Linnaeus, 1758 (Elasmobranchii: Carcharhiniformes

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    Bianca S. Rangel

    Full Text Available ABSTRACT Oviducal gland present in elasmobranchs is correlated to the organism's reproductive strategy, and its functions are to produce mucus, to form the egg's tertiary envelope and to store sperm. The gland contains four zones: club, papillary,baffle and terminal. The structures of the oviduct, oviducal gland and isthmus of blue shark Prionace glauca were described using macroscopic, light microscopy and scanning electron microscopy techniques. The epithelium of the oviduct and isthmus is folded and is a simple, columnar, ciliated lining epithelium with glandular cells. In the oviducal gland, the lining tissues in the four zones are similar to the oviduct and isthmus lining. The terminal zone shows the presence of sperm in the lumen of the secretory tubules, which remains stored even in the absence of recent copulation. Here, these organs were studied and their connections in an attempt to elucidate the mechanisms of reproduction in the blue shark, showing the three-dimensional aspects, thus adding morphological information important for the understanding of the structure and functioning of these organs of fundamental importance in the life of the majority of elasmobranchs.

  7. Begin at the beginning: A BAC-end view of the passion fruit (Passiflora) genome.

    Science.gov (United States)

    Santos, Anselmo Azevedo; Penha, Helen Alves; Bellec, Arnaud; Munhoz, Carla de Freitas; Pedrosa-Harand, Andrea; Bergès, Hélène; Vieira, Maria Lucia Carneiro

    2014-09-26

    The passion fruit (Passiflora edulis) is a tropical crop of economic importance both for juice production and consumption as fresh fruit. The juice is also used in concentrate blends that are consumed worldwide. However, very little is known about the genome of the species. Therefore, improving our understanding of passion fruit genomics is essential and to some degree a pre-requisite if its genetic resources are to be used more efficiently. In this study, we have constructed a large-insert BAC library and provided the first view on the structure and content of the passion fruit genome, using BAC-end sequence (BES) data as a major resource. The library consisted of 82,944 clones and its levels of organellar DNA were very low. The library represents six haploid genome equivalents, and the average insert size was 108 kb. To check its utility for gene isolation, successful macroarray screening experiments were carried out with probes complementary to eight Passiflora gene sequences available in public databases. BACs harbouring those genes were used in fluorescent in situ hybridizations and unique signals were detected for four BACs in three chromosomes (n=9). Then, we explored 10,000 BES and we identified reads likely to contain repetitive mobile elements (19.6% of all BES), simple sequence repeats and putative proteins, and to estimate the GC content (~42%) of the reads. Around 9.6% of all BES were found to have high levels of similarity to plant genes and ontological terms were assigned to more than half of the sequences analysed (940). The vast majority of the top-hits made by our sequences were to Populus trichocarpa (24.8% of the total occurrences), Theobroma cacao (21.6%), Ricinus communis (14.3%), Vitis vinifera (6.5%) and Prunus persica (3.8%). We generated the first large-insert library for a member of Passifloraceae. This BAC library provides a new resource for genetic and genomic studies, as well as it represents a valuable tool for future whole genome

  8. The disappearance of Picea at the end of Upper Pleistocene in the Basque-Cantabrian region: climatic and anthropogenic factors

    International Nuclear Information System (INIS)

    Uzquiano, P.

    1995-01-01

    Charcoal analysis from archaeological sites in Cantabrian region (North of Spain) has yielded evidence of Picea charcoal in Late-Wurmian sediments. Taking into account the behaviour of this taxon in Quaternary climatic cycles, data suggest the presence of small localities of low-density populations near each catchment area of prehistoric settlements. The different function of each site and hunter-gatherers'logistical moves closer to the economic resources are responsible for the presence or absence of Picea, and for the disappearance of its last localities. (author). 16 refs., 4 figs

  9. Comparison of 'time to detection' values between BacT/ALERT VIRTUO and BacT/ALERT 3D instruments for clinical blood culture samples.

    Science.gov (United States)

    Congestrì, Francesco; Pedna, Maria Federica; Fantini, Michela; Samuelli, Michela; Schiavone, Pasqua; Torri, Arianna; Bertini, Stefania; Sambri, Vittorio

    2017-09-01

    The early detection of bacteraemia and fungemia is of paramount importance to guide antimicrobial therapy in septic patients. In this study the 'time to detection' (TTD) value for the new blood culture system BacT/ALERT VIRTUO (VIRTUO) was evaluated in 1462 positive clinical bottles and compared with the TTD for 1601 positive clinical bottles incubated in the BacT/ALERT 3D system (BTA-3D). The most representative microorganisms isolated from bottles incubated in both blood culture systems were divided into eight categories (in order of frequency): coagulase-negative staphylococci (CoNS), Escherichia coli, Enterobacteriaceae (other than E. coli), Staphylococcus aureus, Enterococcus spp, viridans group streptococci, Pseudomonas aeruginosa, and Candida spp. The comparison of TTD values for the two blood culture systems strongly indicated that growth of the first five groups listed above was detected earlier with VIRTUO than with BTA-3D (p culture system can reduce the TTD for more than 75% of isolated microorganisms. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  10. Interference of the Histone Deacetylase Inhibits Pollen Germination and Pollen Tube Growth in Picea wilsonii Mast.

    Directory of Open Access Journals (Sweden)

    Yaning Cui

    Full Text Available Histone deacetylase (HDAC is a crucial component in the regulation of gene expression in various cellular processes in animal and plant cells. HDAC has been reported to play a role in embryogenesis. However, the effect of HDAC on androgamete development remains unclear, especially in gymnosperms. In this study, we used the HDAC inhibitors trichostatin A (TSA and sodium butyrate (NaB to examine the role of HDAC in Picea wilsonii pollen germination and pollen tube elongation. Measurements of the tip-focused Ca2+ gradient revealed that TSA and NaB influenced this gradient. Immunofluorescence showed that actin filaments were disrupted into disorganized fragments. As a result, the vesicle trafficking was disturbed, as determined by FM4-64 labeling. Moreover, the distribution of pectins and callose in cell walls was significantly altered in response to TSA and NaB. Our results suggest that HDAC affects pollen germination and polarized pollen tube growth in Picea wilsonii by affecting the intracellular Ca2+ concentration gradient, actin organization patterns, vesicle trafficking, as well as the deposition and configuration of cell wall components.

  11. Fine roots in stands of Fagus sylvatica and Picea abies along a gradient of soil acidification

    International Nuclear Information System (INIS)

    Braun, Sabine; Cantaluppi, Leonardo; Flueckiger, Walter

    2005-01-01

    Root length of naturally grown young beech trees (Fagus sylvatica L.) was investigated in 26 forest plots of differing base saturation and nitrogen deposition. The relative length of finest roots (<0.25 mm) was found to decrease in soils with low base saturation. A similar reduction of finest roots in plots with high nitrogen deposition was masked by the effect of base saturation. The formation of adventitious roots was enhanced in acidic soils. The analysis of 128 soil profiles for fine roots of all species present in stands of either Fagus sylvatica L., Picea abies [Karst.] L. or both showed a decreased rooting depth in soils with ≤20% base saturation and in hydromorphic soils. For base rich, well drained soils an average rooting depth of 108 cm was found. This decreased by 28 cm on acidic, well drained soils. The results suggest an effect of the current soil acidification in Switzerland and possibly also of nitrogen deposition on the fine root systems of forest trees. - Fine root length of Fagus sylvatica and fine root depth in stands of Fagus sylvatica and/or Picea abies were impaired in soils with low base saturation

  12. BAC-end sequence-based SNPs and Bin mapping for rapid integration of physical and genetic maps in apple.

    Science.gov (United States)

    Han, Yuepeng; Chagné, David; Gasic, Ksenija; Rikkerink, Erik H A; Beever, Jonathan E; Gardiner, Susan E; Korban, Schuyler S

    2009-03-01

    A genome-wide BAC physical map of the apple, Malus x domestica Borkh., has been recently developed. Here, we report on integrating the physical and genetic maps of the apple using a SNP-based approach in conjunction with bin mapping. Briefly, BAC clones located at ends of BAC contigs were selected, and sequenced at both ends. The BAC end sequences (BESs) were used to identify candidate SNPs. Subsequently, these candidate SNPs were genetically mapped using a bin mapping strategy for the purpose of mapping the physical onto the genetic map. Using this approach, 52 (23%) out of 228 BESs tested were successfully exploited to develop SNPs. These SNPs anchored 51 contigs, spanning approximately 37 Mb in cumulative physical length, onto 14 linkage groups. The reliability of the integration of the physical and genetic maps using this SNP-based strategy is described, and the results confirm the feasibility of this approach to construct an integrated physical and genetic maps for apple.

  13. Development of Chromosome-Specific BAC Resources for Genomics of Bread Wheat

    Czech Academy of Sciences Publication Activity Database

    Šafář, Jan; Šimková, Hana; Kubaláková, Marie; Čihalíková, Jarmila; Suchánková, Pavla; Bartoš, Jan; Doležel, Jaroslav

    2010-01-01

    Roč. 129, 1-3 (2010), s. 211-223 ISSN 1424-8581 R&D Projects: GA ČR GA521/07/1573; GA MŠk(CZ) LC06004 Grant - others:European Community’s Seventh Framework Programme(XE) FP7/2007–2013 Institutional research plan: CEZ:AV0Z50380511 Keywords : BAC library * Chromosome * DNA markers Subject RIV: GE - Plant Breeding Impact factor: 1.783, year: 2010

  14. Chromosomal mapping of canine-derived BAC clones to the red fox and American mink genomes.

    Science.gov (United States)

    Kukekova, Anna V; Vorobieva, Nadegda V; Beklemisheva, Violetta R; Johnson, Jennifer L; Temnykh, Svetlana V; Yudkin, Dmitry V; Trut, Lyudmila N; Andre, Catherine; Galibert, Francis; Aguirre, Gustavo D; Acland, Gregory M; Graphodatsky, Alexander S

    2009-01-01

    High-quality sequencing of the dog (Canis lupus familiaris) genome has enabled enormous progress in genetic mapping of canine phenotypic variation. The red fox (Vulpes vulpes), another canid species, also exhibits a wide range of variation in coat color, morphology, and behavior. Although the fox genome has not yet been sequenced, canine genomic resources have been used to construct a meiotic linkage map of the red fox genome and begin genetic mapping in foxes. However, a more detailed gene-specific comparative map between the dog and fox genomes is required to establish gene order within homologous regions of dog and fox chromosomes and to refine breakpoints between homologous chromosomes of the 2 species. In the current study, we tested whether canine-derived gene-containing bacterial artificial chromosome (BAC) clones can be routinely used to build a gene-specific map of the red fox genome. Forty canine BAC clones were mapped to the red fox genome by fluorescence in situ hybridization (FISH). Each clone was uniquely assigned to a single fox chromosome, and the locations of 38 clones agreed with cytogenetic predictions. These results clearly demonstrate the utility of FISH mapping for construction of a whole-genome gene-specific map of the red fox. The further possibility of using canine BAC clones to map genes in the American mink (Mustela vison) genome was also explored. Much lower success was obtained for this more distantly related farm-bred species, although a few BAC clones were mapped to the predicted chromosomal locations.

  15. Establishing gene models from the Pinus pinaster genome using gene capture and BAC sequencing.

    Science.gov (United States)

    Seoane-Zonjic, Pedro; Cañas, Rafael A; Bautista, Rocío; Gómez-Maldonado, Josefa; Arrillaga, Isabel; Fernández-Pozo, Noé; Claros, M Gonzalo; Cánovas, Francisco M; Ávila, Concepción

    2016-02-27

    In the era of DNA throughput sequencing, assembling and understanding gymnosperm mega-genomes remains a challenge. Although drafts of three conifer genomes have recently been published, this number is too low to understand the full complexity of conifer genomes. Using techniques focused on specific genes, gene models can be established that can aid in the assembly of gene-rich regions, and this information can be used to compare genomes and understand functional evolution. In this study, gene capture technology combined with BAC isolation and sequencing was used as an experimental approach to establish de novo gene structures without a reference genome. Probes were designed for 866 maritime pine transcripts to sequence genes captured from genomic DNA. The gene models were constructed using GeneAssembler, a new bioinformatic pipeline, which reconstructed over 82% of the gene structures, and a high proportion (85%) of the captured gene models contained sequences from the promoter regulatory region. In a parallel experiment, the P. pinaster BAC library was screened to isolate clones containing genes whose cDNA sequence were already available. BAC clones containing the asparagine synthetase, sucrose synthase and xyloglucan endotransglycosylase gene sequences were isolated and used in this study. The gene models derived from the gene capture approach were compared with the genomic sequences derived from the BAC clones. This combined approach is a particularly efficient way to capture the genomic structures of gene families with a small number of members. The experimental approach used in this study is a valuable combined technique to study genomic gene structures in species for which a reference genome is unavailable. It can be used to establish exon/intron boundaries in unknown gene structures, to reconstruct incomplete genes and to obtain promoter sequences that can be used for transcriptional studies. A bioinformatics algorithm (GeneAssembler) is also provided as a

  16. Cytogenetic Analysis of Populus trichocarpa - Ribosomal DNA, Telomere Repeat Sequence, and Marker-selected BACs

    Science.gov (United States)

    M.N. lslam-Faridi; C.D. Nelson; S.P. DiFazio; L.E. Gunter; G.A. Tuskan

    2009-01-01

    The 185-285 rDNA and 55 rDNA loci in Populus trichocarpa were localized using fluorescent in situ hybridization (FISH). Two 185-285 rDNA sites and one 55 rDNA site were identified and located at the ends of 3 different chromosomes. FISH signals from the Arabidopsis-type telomere repeat sequence were observed at the distal ends of each chromosome. Six BAC clones...

  17. Versatile P(acman) BAC Libraries for Transgenesis Studies in Drosophila melanogaster

    Energy Technology Data Exchange (ETDEWEB)

    Venken, Koen J.T.; Carlson, Joseph W.; Schulze, Karen L.; Pan, Hongling; He, Yuchun; Spokony, Rebecca; Wan, Kenneth H.; Koriabine, Maxim; de Jong, Pieter J.; White, Kevin P.; Bellen, Hugo J.; Hoskins, Roger A.

    2009-04-21

    We constructed Drosophila melanogaster BAC libraries with 21-kb and 83-kb inserts in the P(acman) system. Clones representing 12-fold coverage and encompassing more than 95percent of annotated genes were mapped onto the reference genome. These clones can be integrated into predetermined attP sites in the genome using Phi C31 integrase to rescue mutations. They can be modified through recombineering, for example to incorporate protein tags and assess expression patterns.

  18. A BAC-based physical map of the Drosophila buzzatii genome

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez, Josefa; Nefedov, Michael; Bosdet, Ian; Casals, Ferran; Calvete, Oriol; Delprat, Alejandra; Shin, Heesun; Chiu, Readman; Mathewson, Carrie; Wye, Natasja; Hoskins, Roger A.; Schein, JacquelineE.; de Jong, Pieter; Ruiz, Alfredo

    2005-03-18

    Large-insert genomic libraries facilitate cloning of large genomic regions, allow the construction of clone-based physical maps and provide useful resources for sequencing entire genomes. Drosophilabuzzatii is a representative species of the repleta group in the Drosophila subgenus, which is being widely used as a model in studies of genome evolution, ecological adaptation and speciation. We constructed a Bacterial Artificial Chromosome (BAC) genomic library of D. buzzatii using the shuttle vector pTARBAC2.1. The library comprises 18,353 clones with an average insert size of 152 kb and a {approx}18X expected representation of the D. buzzatii euchromatic genome. We screened the entire library with six euchromatic gene probes and estimated the actual genome representation to be {approx}23X. In addition, we fingerprinted by restriction digestion and agarose gel electrophoresis a sample of 9,555 clones, and assembled them using Finger Printed Contigs (FPC) software and manual editing into 345 contigs (mean of 26 clones per contig) and 670singletons. Finally, we anchored 181 large contigs (containing 7,788clones) to the D. buzzatii salivary gland polytene chromosomes by in situ hybridization of 427 representative clones. The BAC library and a database with all the information regarding the high coverage BAC-based physical map described in this paper are available to the research community.

  19. Study on the Mitochondrial Genome of Sea Island Cotton (Gossypium barbadense) by BAC Library Screening

    Institute of Scientific and Technical Information of China (English)

    SU Ai-guo; LI Shuang-shuang; LIU Guo-zheng; LEI Bin-bin; KANG Ding-ming; LI Zhao-hu; MA Zhi-ying; HUA Jin-ping

    2014-01-01

    The plant mitochondrial genome displays complex features, particularly in terms of cytoplasmic male sterility (CMS). Therefore, research on the cotton mitochondrial genome may provide important information for analyzing genome evolution and exploring the molecular mechanism of CMS. In this paper, we present a preliminary study on the mitochondrial genome of sea island cotton (Gossypium barbadense) based on positive clones from the bacterial artiifcial chromosome (BAC) library. Thirty-ifve primers designed with the conserved sequences of functional genes and exons of mitochondria were used to screen positive clones in the genome library of the sea island cotton variety called Pima 90-53. Ten BAC clones were obtained and veriifed for further study. A contig was obtained based on six overlapping clones and subsequently laid out primarily on the mitochondrial genome. One BAC clone, clone 6 harbored with the inserter of approximate 115 kb mtDNA sequence, in which more than 10 primers fragments could be ampliifed, was sequenced and assembled using the Solexa strategy. Fifteen mitochondrial functional genes were revealed in clone 6 by gene annotation. The characteristics of the syntenic gene/exon of the sequences and RNA editing were preliminarily predicted.

  20. A BAC-based transgenic mouse specifically expresses an inducible Cre in the urothelium.

    Directory of Open Access Journals (Sweden)

    Tian Huai Shen

    Full Text Available Cre-loxp mediated conditional knockout strategy has played critical roles for revealing functions of many genes essential for development, as well as the causal relationships between gene mutations and diseases in the postnatal adult mice. One key factor of this strategy is the availability of mice with tissue- or cell type-specific Cre expression. However, the success of the traditional molecular cloning approach to generate mice with tissue specific Cre expression often depends on luck. Here we provide a better alternative by using bacterial artificial chromosome (BAC-based recombineering to insert iCreERT2 cDNA at the ATG start of the Upk2 gene. The BAC-based transgenic mice express the inducible Cre specifically in the urothelium as demonstrated by mRNA expression and staining for LacZ expression after crossing with a Rosa26 reporter mouse. Taking into consideration the size of the gene of interest and neighboring genes included in a BAC, this method should be widely applicable for generation of mice with tissue specific gene expression or deletions in a more specific manner than previously reported.

  1. Construction of a 7-fold BAC library and cytogenetic mapping of 10 genes in the giant panda (Ailuropoda melanoleuca)

    OpenAIRE

    Liu, Wei; Zhao, Yonghui; Liu, Zhaoliang; Zhang, Ying; Lian, Zhengxing; Li, Ning

    2006-01-01

    Abstract Background The giant panda, one of the most primitive carnivores, is an endangered animal. Although it has been the subject of many interesting studies during recent years, little is known about its genome. In order to promote research on this genome, a bacterial artificial chromosome (BAC) library of the giant panda was constructed in this study. Results This BAC library contains 198,844 clones with an average insert size of 108 kb, which represents approximately seven equivalents o...

  2. A first generation BAC-based physical map of the rainbow trout genome

    Directory of Open Access Journals (Sweden)

    Thorgaard Gary H

    2009-10-01

    Full Text Available Abstract Background Rainbow trout (Oncorhynchus mykiss are the most-widely cultivated cold freshwater fish in the world and an important model species for many research areas. Coupling great interest in this species as a research model with the need for genetic improvement of aquaculture production efficiency traits justifies the continued development of genomics research resources. Many quantitative trait loci (QTL have been identified for production and life-history traits in rainbow trout. A bacterial artificial chromosome (BAC physical map is needed to facilitate fine mapping of QTL and the selection of positional candidate genes for incorporation in marker-assisted selection (MAS for improving rainbow trout aquaculture production. This resource will also facilitate efforts to obtain and assemble a whole-genome reference sequence for this species. Results The physical map was constructed from DNA fingerprinting of 192,096 BAC clones using the 4-color high-information content fingerprinting (HICF method. The clones were assembled into physical map contigs using the finger-printing contig (FPC program. The map is composed of 4,173 contigs and 9,379 singletons. The total number of unique fingerprinting fragments (consensus bands in contigs is 1,185,157, which corresponds to an estimated physical length of 2.0 Gb. The map assembly was validated by 1 comparison with probe hybridization results and agarose gel fingerprinting contigs; and 2 anchoring large contigs to the microsatellite-based genetic linkage map. Conclusion The production and validation of the first BAC physical map of the rainbow trout genome is described in this paper. We are currently integrating this map with the NCCCWA genetic map using more than 200 microsatellites isolated from BAC end sequences and by identifying BACs that harbor more than 300 previously mapped markers. The availability of an integrated physical and genetic map will enable detailed comparative genome

  3. A highly redundant BAC library of Atlantic salmon (Salmo salar: an important tool for salmon projects

    Directory of Open Access Journals (Sweden)

    Koop Ben F

    2005-04-01

    Full Text Available Abstract Background As farming of Atlantic salmon is growing as an aquaculture enterprise, the need to identify the genomic mechanisms for specific traits is becoming more important in breeding and management of the animal. Traits of importance might be related to growth, disease resistance, food conversion efficiency, color or taste. To identify genomic regions responsible for specific traits, genomic large insert libraries have previously proven to be of crucial importance. These large insert libraries can be screened using gene or genetic markers in order to identify and map regions of interest. Furthermore, large-scale mapping can utilize highly redundant libraries in genome projects, and hence provide valuable data on the genome structure. Results Here we report the construction and characterization of a highly redundant bacterial artificial chromosome (BAC library constructed from a Norwegian aquaculture strain male of Atlantic salmon (Salmo salar. The library consists of a total number of 305 557 clones, in which approximately 299 000 are recombinants. The average insert size of the library is 188 kbp, representing 18-fold genome coverage. High-density filters each consisting of 18 432 clones spotted in duplicates have been produced for hybridization screening, and are publicly available 1. To characterize the library, 15 expressed sequence tags (ESTs derived overgos and 12 oligo sequences derived from microsatellite markers were used in hybridization screening of the complete BAC library. Secondary hybridizations with individual probes were performed for the clones detected. The BACs positive for the EST probes were fingerprinted and mapped into contigs, yielding an average of 3 contigs for each probe. Clones identified using genomic probes were PCR verified using microsatellite specific primers. Conclusion Identification of genes and genomic regions of interest is greatly aided by the availability of the CHORI-214 Atlantic salmon BAC

  4. Generation of a BAC-based physical map of the melon genome

    Directory of Open Access Journals (Sweden)

    Puigdomènech Pere

    2010-05-01

    Full Text Available Abstract Background Cucumis melo (melon belongs to the Cucurbitaceae family, whose economic importance among horticulture crops is second only to Solanaceae. Melon has high intra-specific genetic variation, morphologic diversity and a small genome size (450 Mb, which make this species suitable for a great variety of molecular and genetic studies that can lead to the development of tools for breeding varieties of the species. A number of genetic and genomic resources have already been developed, such as several genetic maps and BAC genomic libraries. These tools are essential for the construction of a physical map, a valuable resource for map-based cloning, comparative genomics and assembly of whole genome sequencing data. However, no physical map of any Cucurbitaceae has yet been developed. A project has recently been started to sequence the complete melon genome following a whole-genome shotgun strategy, which makes use of massive sequencing data. A BAC-based melon physical map will be a useful tool to help assemble and refine the draft genome data that is being produced. Results A melon physical map was constructed using a 5.7 × BAC library and a genetic map previously developed in our laboratories. High-information-content fingerprinting (HICF was carried out on 23,040 BAC clones, digesting with five restriction enzymes and SNaPshot labeling, followed by contig assembly with FPC software. The physical map has 1,355 contigs and 441 singletons, with an estimated physical length of 407 Mb (0.9 × coverage of the genome and the longest contig being 3.2 Mb. The anchoring of 845 BAC clones to 178 genetic markers (100 RFLPs, 76 SNPs and 2 SSRs also allowed the genetic positioning of 183 physical map contigs/singletons, representing 55 Mb (12% of the melon genome, to individual chromosomal loci. The melon FPC database is available for download at http://melonomics.upv.es/static/files/public/physical_map/. Conclusions Here we report the construction

  5. Susceptibility of burned black spruce (Picea mariana) forests to non-native plant invasions in interior Alaska

    Science.gov (United States)

    Katie V. Spellman; Christa P.H. Mulder; Teresa N. Hollingsworth

    2014-01-01

    As climate rapidly warms at high-latitudes, the boreal forest faces the simultaneous threats of increasing invasive plant abundances and increasing area burned by wildfire. Highly flammable and widespread black spruce (Picea mariana) forest represents a boreal habitat that may be increasingly susceptible to non-native plant invasion. This study assess the role of burn...

  6. CITOGENETICS EFFECTS INDUCED BY THE ASCORBIC ACID TREATMENT OF LARIX DECIDUA MILL. SSP. CARPATICA AND PICEA ABIES (L. KARST

    Directory of Open Access Journals (Sweden)

    Ioana Ieremia

    2006-08-01

    Full Text Available The paper present the influence of ascorbic acid upon the mitotic division of Larix decidua Mill ssp. carpatica and Picea abies (L. Karst. The treatment is applied of two variants, germinated seed in ascorbic acid (variantAand germinated seeds in disttilate water, than treated with ascorbic acid in 3 concentrations (variant B.

  7. Cestodes of the blue shark, Prionace glauca (Linnaeus 1758), (Carcharhiniformes: Carcharhinidae), off the west coast of Baja California Sur, Mexico.

    Science.gov (United States)

    Méndez, Oscar; Galván-Magaña, Felipe

    2016-03-03

    The cestode species recovered from the spiral intestines of 27 blue sharks (Prionace glauca) (Linnaeus, 1758) are reported from the western coast of Baja California Sur (BCS). The sampling was undertaken on a monthly basis from January 2003 to January 2004. The helminthological examination indicated the presence of four species of cestodes: Platybothrium auriculatum Yamaguti, 1952; Prosobothrium japonicum Yamaguti, 1934; Anthobothrium caseyi (Yamaguti, 1934) Ruhnke & Caira, 2009; and Paraorygmatobothrium prionacis (Yamaguti, 1934) Ruhnke, 1994. Of all the 27 sharks examined, 88.8% were infected with at least one cestode species. The most frequent species was P. auriculatum infecting 85% of the spiral intestines examined. In contrast the species with the highest mean intensity was P. prionacis (80.4 200). The species richness of cestodes in P. glauca is very similar in other regions of the world despite its wide distribution; however, this richness is low compared with other species of sharks within the same family. The feeding and host-specific are important factors that influence the parameters of infection of cestodes in this shark. On the west coast of BCS, Prionace glauca feeds mainly on red crab Pleuroncodes planipes Stimpson, 1860; squids Gonatus californiensis Young, 1972, Ancistrocheirus lesueurii (D'Orbigny, 1842), Haliphron atlanticus Steenstrup, 1861, and low proportion of fish teleosts as Merluccius productus (Ayres, 1855), Sardinops sp. Hubbs, 1929 and Scomber japonicus Houttuyn, 1872. We speculate that these prey could be involved as the second intermediate hosts of these cestodes, as in other members of these genera, although the life cycles of none are known.

  8. Identification and characterization of polymorphic microsatellite loci in the blue shark Prionace glauca, and cross-amplification in other shark species.

    Science.gov (United States)

    Mendonça, F F; Ussami, L H F; Hashimoto, D T; Pereira, L H G; Porto-Foresti, F; Oliveira, C; Gadig, O B F; Foresti, F

    2012-06-01

    Two to 14 alleles were found to be segregating per locus (mean 5·2), with observed and expected heterozygosities ranging from 0·08 to 0·78 and 0·08 to 0·94, respectively. Cross-amplification of six of these microsatellite loci indicated that they are also polymorphic in three species of Carcharhiniformes and two species of Lamniformes. The newly developed primers reported here constitute a useful tool for genetic population analyses on Prionace glauca and, potentially, other related species. © 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.

  9. Construction of an American mink Bacterial Artificial Chromosome (BAC library and sequencing candidate genes important for the fur industry

    Directory of Open Access Journals (Sweden)

    Christensen Knud

    2011-07-01

    Full Text Available Abstract Background Bacterial artificial chromosome (BAC libraries continue to be invaluable tools for the genomic analysis of complex organisms. Complemented by the newly and fast growing deep sequencing technologies, they provide an excellent source of information in genomics projects. Results Here, we report the construction and characterization of the CHORI-231 BAC library constructed from a Danish-farmed, male American mink (Neovison vison. The library contains approximately 165,888 clones with an average insert size of 170 kb, representing approximately 10-fold coverage. High-density filters, each consisting of 18,432 clones spotted in duplicate, have been produced for hybridization screening and are publicly available. Overgo probes derived from expressed sequence tags (ESTs, representing 21 candidate genes for traits important for the mink industry, were used to screen the BAC library. These included candidate genes for coat coloring, hair growth and length, coarseness, and some receptors potentially involved in viral diseases in mink. The extensive screening yielded positive results for 19 of these genes. Thirty-five clones corresponding to 19 genes were sequenced using 454 Roche, and large contigs (184 kb in average were assembled. Knowing the complete sequences of these candidate genes will enable confirmation of the association with a phenotype and the finding of causative mutations for the targeted phenotypes. Additionally, 1577 BAC clones were end sequenced; 2505 BAC end sequences (80% of BACs were obtained. An excess of 2 Mb has been analyzed, thus giving a snapshot of the mink genome. Conclusions The availability of the CHORI-321 American mink BAC library will aid in identification of genes and genomic regions of interest. We have demonstrated how the library can be used to identify specific genes of interest, develop genetic markers, and for BAC end sequencing and deep sequencing of selected clones. To our knowledge, this is the

  10. Validation of the French version of the BACS (the brief assessment of cognition in schizophrenia) among 50 French schizophrenic patients.

    Science.gov (United States)

    Bralet, Marie-Cécile; Falissard, Bruno; Neveu, Xavier; Lucas-Ross, Margaret; Eskenazi, Anne-Marie; Keefe, Richard S E

    2007-09-01

    Schizophrenic patients demonstrate impairments in several key dimensions of cognition. These impairments are correlated with important aspects of functional outcome. While assessment of these cognition disorders is increasingly becoming a part of clinical and research practice in schizophrenia, there is no standard and easily administered test battery. The BACS (Brief Assessment of Cognition in Schizophrenia) has been validated in English language [Keefe RSE, Golberg TE, Harvey PD, Gold JM, Poe MP, Coughenour L. The Brief Assessment of Cognition in Schizophrenia: reliability, sensibility, and comparison with a standard neurocognitive battery. Schizophr. Res 2004;68:283-97], and was found to be as sensitive to cognitive dysfunction as a standard battery of tests, with the advantage of requiring less than 35 min to complete. We developed a French adaptation of the BACS and this study tested its ease of administration and concurrent validity. Correlation analyses between the BACS (version A) and a standard battery were performed. A sample of 50 stable schizophrenic patients received the French Version A of the BACS in a first session, and in a second session a standard battery. All the patients completed each of the subtests of the French BACS . The mean duration of completion for the BACS French version was 36 min (S.D.=5.56). A correlation analysis between the BACS (version A) global score and the standard battery global score showed a significant result (r=0.81, p<0.0001). The correlation analysis between the BACS (version A) sub-scores and the standard battery sub-scores showed significant results for verbal memory, working memory, verbal fluency, attention and speed of information processing and executive functions (p<0.001) and for motor speed (p<0.05). The French Version of the BACS is easier to use in French schizophrenic patients compared to a standard battery (administration shorter and completion rate better) and its good psychometric properties suggest

  11. An experimental study on the effects of exhaust gas on spruce (Picea abies L. Karst.)

    Energy Technology Data Exchange (ETDEWEB)

    Hautala, E.L.; Holopainen, J.; Kaerenlampi, L. [Kuopio Univ. (Finland). Dept. of Ecology and Environmental Science; Surakka, J.; Ruuskanen, J. [Kuopio Univ. (Finland). Dept. of Environmental Sciences

    1995-12-31

    Motor vehicle exhausts are significant contributors to air pollution. Besides fine particles and inorganic gases, like CO, SO{sub 2} and NO{sub x}, exhaust gas contains a large group of aromatic hydrocarbon compounds, many of which are phytotoxic. In field studies, exhausts are found to have both direct and indirect harmful effects on roadside plants. However, only few experimental studies have been made about the effects of exhaust gas emissions on coniferous trees. The aim of this study was to survey the effects of exhausts on spruce (Picea abies L. Karst.) in standardized conditions. The concentrations of major exhaust gas components in the chamber atmosphere were detected simultaneously. The effects of exhaust on epistomatal waxes of first-year spruce needles are described. (author)

  12. An experimental study on the effects of exhaust gas on spruce (Picea abies L. Karst.)

    Energy Technology Data Exchange (ETDEWEB)

    Hautala, E L; Holopainen, J; Kaerenlampi, L [Kuopio Univ. (Finland). Dept. of Ecology and Environmental Science; Surakka, J; Ruuskanen, J [Kuopio Univ. (Finland). Dept. of Environmental Sciences

    1996-12-31

    Motor vehicle exhausts are significant contributors to air pollution. Besides fine particles and inorganic gases, like CO, SO{sub 2} and NO{sub x}, exhaust gas contains a large group of aromatic hydrocarbon compounds, many of which are phytotoxic. In field studies, exhausts are found to have both direct and indirect harmful effects on roadside plants. However, only few experimental studies have been made about the effects of exhaust gas emissions on coniferous trees. The aim of this study was to survey the effects of exhausts on spruce (Picea abies L. Karst.) in standardized conditions. The concentrations of major exhaust gas components in the chamber atmosphere were detected simultaneously. The effects of exhaust on epistomatal waxes of first-year spruce needles are described. (author)

  13. GENERAL ASSESMENT OF SEED STANDS OF ORIENTAL SPRUCE (Picea orientalis L. Link.

    Directory of Open Access Journals (Sweden)

    İbrahim Turna

    2001-04-01

    Full Text Available The seeds for afforestation studies with oriental spruce (Picea orientalis L.Link. populations are usually gathered from already established seed stands. This study showed that most of the seed stands were not selected appropriately and some of them were not properly protected and maintained when evaluated according to the “criteria for selection and management of seed stands”. The average slope of seed stands was as high as 43% and the average size varied from 33.5 ha to 147.5 ha Since the stands were open to social disturbances and wind effects, the number of trees per ha decreased 51% and the number of seed trees dropped to a level of 56%. In conclusion, seeds collected from poorly established seed stands will likely jeopardize the success in reforestation studies in the future.

  14. Photolysis of polycyclic aromatic hydrocarbons adsorbed on spruce [Picea abies (L.) Karst.] needles under sunlight irradiation

    International Nuclear Information System (INIS)

    Niu Junfeng; Chen Jingwen; Martens, D.; Quan Xie; Yang Fenglin; Kettrup, A.; Schramm, K.-W.

    2003-01-01

    Photolysis of PAHs on surfaces may determine their ultimate fate in the environment. - Photolysis of polycyclic aromatic hydrocarbons (PAHs) sorbed on surfaces of spruce [Picea abies (L.) Karst.] needles under sunlight irradiation was investigated. PAHs were produced by combustion of polyvinyl chloride (PVC), wood, high-density polyethylene (HDPE), and styrene in a stove. The factors of sunlight irradiation on the surfaces of spruce needles were taken into consideration when investigating the kinetic parameters. The photolysis of the 18 PAHs under study follows first-order kinetics. The photolysis half-lives range from 15 h for dibenzo(a,h)anthracene to 75 h for phenanthrene. Photolysis of some PAHs on surfaces of spruce needles may play an important role on the fate of PAHs in the environment

  15. Toward a molecular cytogenetic map for cultivated sunflower (Helianthus annuus L.) by landed BAC/BIBAC clones.

    Science.gov (United States)

    Feng, Jiuhuan; Liu, Zhao; Cai, Xiwen; Jan, Chao-Chien

    2013-01-01

    Conventional karyotypes and various genetic linkage maps have been established in sunflower (Helianthus annuus L., 2n = 34). However, the relationship between linkage groups and individual chromosomes of sunflower remains unknown and has considerable relevance for the sunflower research community. Recently, a set of linkage group-specific bacterial /binary bacterial artificial chromosome (BAC/BIBAC) clones was identified from two complementary BAC and BIBAC libraries constructed for cultivated sunflower cv. HA89. In the present study, we used these linkage group-specific clones (~100 kb in size) as probes to in situ hybridize to HA89 mitotic chromosomes at metaphase using the BAC-fluorescence in situ hybridization (FISH) technique. Because a characteristic of the sunflower genome is the abundance of repetitive DNA sequences, a high ratio of blocking DNA to probe DNA was applied to hybridization reactions to minimize the background noise. As a result, all sunflower chromosomes were anchored by one or two BAC/BIBAC clones with specific FISH signals. FISH analysis based on tandem repetitive sequences, such as rRNA genes, has been previously reported; however, the BAC-FISH technique developed here using restriction fragment length polymorphism (RFLP)-derived BAC/BIBAC clones as probes to apply genome-wide analysis is new for sunflower. As chromosome-specific cytogenetic markers, the selected BAC/BIBAC clones that encompass the 17 linkage groups provide a valuable tool for identifying sunflower cytogenetic stocks (such as trisomics) and tracking alien chromosomes in interspecific crosses. This work also demonstrates the potential of using a large-insert DNA library for the development of molecular cytogenetic resources.

  16. The first generation of a BAC-based physical map of Brassica rapa

    Directory of Open Access Journals (Sweden)

    Lee Soo

    2008-06-01

    Full Text Available Abstract Background The genus Brassica includes the most extensively cultivated vegetable crops worldwide. Investigation of the Brassica genome presents excellent challenges to study plant genome evolution and divergence of gene function associated with polyploidy and genome hybridization. A physical map of the B. rapa genome is a fundamental tool for analysis of Brassica "A" genome structure. Integration of a physical map with an existing genetic map by linking genetic markers and BAC clones in the sequencing pipeline provides a crucial resource for the ongoing genome sequencing effort and assembly of whole genome sequences. Results A genome-wide physical map of the B. rapa genome was constructed by the capillary electrophoresis-based fingerprinting of 67,468 Bacterial Artificial Chromosome (BAC clones using the five restriction enzyme SNaPshot technique. The clones were assembled into contigs by means of FPC v8.5.3. After contig validation and manual editing, the resulting contig assembly consists of 1,428 contigs and is estimated to span 717 Mb in physical length. This map provides 242 anchored contigs on 10 linkage groups to be served as seed points from which to continue bidirectional chromosome extension for genome sequencing. Conclusion The map reported here is the first physical map for Brassica "A" genome based on the High Information Content Fingerprinting (HICF technique. This physical map will serve as a fundamental genomic resource for accelerating genome sequencing, assembly of BAC sequences, and comparative genomics between Brassica genomes. The current build of the B. rapa physical map is available at the B. rapa Genome Project website for the user community.

  17. Enhanced biodegradation of petrochemical wastewater using ozonation and BAC advanced treatment system

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chi-Kang; Tsai, Tsung-Yueh; Liu, Jiunn-Ching; Chen, Mei-Chen [Energy and Resources Labs., ITRI, Hsinchu (Taiwan)

    2001-07-01

    The characteristics of degradation/conversion of bio-refractory and the growth of a biofilm are investigated in laboratory-scale pre-ozonation and lifted moving-bed biological activated carbon (BAC) advanced treatment processes treating phenol, benzoic acid, aminobenzoic acid and petrochemical industry wastewater which contains acrylonitrile butadiene styrene (ABS). The optimal reaction time and ozone dosage of pre-ozonation for bio-refractory conversion were determined to be 30 min and 100-200mg O{sub 3}/hr, respectively. After pre-ozonation of 30 min treatment, BOD{sub 5}/COD ratio of influent and effluent increased apparently from 20 to 35%, approximately. However, the change of pH in pre-ozonation was inconspicuous. The optimal flow rate of influent and air were controlled at 1.6 1/h and 120-l50nl/min in lifted moving-bed BAC advanced treatment reactor. A COD removal efficiency of 85-95% and 70-90% may be maintained by using an organic loading of 3.2-6.3kg COD/m{sup 3} day and 0.6-1.6 kg-COD/m{sup 3} day with an HRT of 6.0 h as secondary and advanced treatment system, respectively. The time required for the BAC bed to be regenerated by a thermal regeneration is prolonged 4-5 times more than that of GAC system. It can be estimated that the enhanced COD removal capability of the biofilm was not only due to the increase in the COD removal capability of acclimated bacteria, but also due to species succession of bacteria in bio-film ecosystem. (Author)

  18. Experimental tolerance to boron of the plant species Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea in Argentina

    International Nuclear Information System (INIS)

    Viana, Marta L. de; Albarracin Franco, Silvia

    2008-01-01

    The activity of the borate deposits industries constitutes a point source and diffuse pollution of air, soil and water. Therefore, the study and experimentation on possible ways to offset this impact is a priority. A relatively new technique to decontaminate soils is phytoremediation, which uses plants and associated microorganisms. The first step is to identify tolerant plant species, which is the focus of this work. An experiment was conducted in the laboratory to evaluate the germination, survival and growth of different species in different concentrations of boron. At the beginning and end of the experiment was determined concentration of boron in the substrate for each treatment and for substrates with and without vegetation. Significant differences due to treatment, the species and species-treatment interaction. M. sativa, N. glauca and J. mimosifolia were the species most tolerant to boron. The other species showed a decrease in all variables-response function of the concentration of the contaminant. All had low survival in the highest concentration. The decrease of boron was highest in the treatment of 30 ppm of boron with M. sativa and the lowest was recorded in the treatment of 20 ppm of boron with J. mimosifolia and 30 ppm of boron with T. stans and S. oleracea. It is concluded that N. glauca, M. sativa and J. mimosifolia could be considered as promising remediation. (author) [es

  19. Elevated ozone negatively affects photosynthesis of current-year leaves but not previous-year leaves in evergreen Cyclobalanopsis glauca seedlings

    International Nuclear Information System (INIS)

    Zhang, Weiwei; Feng, Zhaozhong; Wang, Xiaoke; Niu, Junfeng

    2014-01-01

    To assess the effects of leaf age/layer on the response of photosynthesis to chronic ozone (O 3 ), Cyclobalanopsis glauca seedlings, a dominant evergreen broadleaf tree species in sub-tropical regions, were exposed to either ambient air (AA) or elevated O 3 (AA + 60 ppb O 3 , E-O 3 ) for two growing seasons in open-top chambers. Chlorophyll content, gas exchange and chlorophyll a fluorescence were investigated three times throughout the 2nd year of O 3 exposure. Results indicated that E-O 3 decreased photosynthetic parameters, particularly light-saturated photosynthesis rate, stomatal conductance and effective quantum yield of PSII photochemistry of current-year leaves but not previous-year leaves. Stomatal conductance of plants grown under ambient conditions partially contributed to the different response to E-O 3 between leaf layers. Light radiation or other physiological and biochemical processes closely related to photosynthesis might play important roles. All suggested that leaf ages or layers should be considered when assessing O 3 risk on evergreen woody species. -- Highlights: • Response of evergreen Cyclobalanopsis glauca to O 3 was investigated. • Elevated O 3 significantly reduced photosynthesis of current-year leaves. • Previous-year leaves showed little response to O 3 . • Stomatal conductance contributes to the response difference to O 3 among leaf ages. -- Impacts of elevated O 3 on photosynthesis of evergreen woody species depend on leaf ages

  20. Effect of wildfire on soil physical and chemical properties in a Nothofagus glauca forest, Chile Efecto del fuego en las propiedades físicas y químicas en un bosque de Nothofagus glauca en Chile

    Directory of Open Access Journals (Sweden)

    CREIGHTON M. LITTON

    2003-12-01

    Full Text Available Effects of a wildfire on soil chemical and physical properties in a Nothofagus glauca (Phil. Krasser forest in the Coastal Mountain Range of south-central Chile were investigated. Response of the soil during the first two years following a wildfire was examined, where data from soil in a burned forest were compared to that in an adjacent, unburned stand. The effects that wildfire have on soil properties in this highly fragmented ecosystem are not well understood, but results from this study suggest similar responses to those found in other mediterranean forest systems. Both physical (bulk density, percent soil moisture, and soil organic matter content and chemical properties (exchangeable inorganic nitrogen, extractable phosphorus, exchangeable potassium, and soil pH were examined, and data presented here suggest that soil properties vary in their initial response to fire in this ecosystem. Soil organic matter content and soil moisture decreased following fire and remained lower than values from unburned plots for the duration of the study. Exchangeable potassium increased initially after burning, but values in burned plots decreased with time and by the end of two years were significantly lower than in unburned soil. In turn, extractable phosphorus and soil pH both increased immediately following wildfire and values in burned plots remained significantly higher than unburned plots for the entire measurement period. Exchangeable inorganic nitrogen reached higher levels in soil of burned plots for the autumn measurements (April 1997 and 1998 and lower values in burned plots for the spring measurements (November 1997 and 1998. Soil bulk density remained unchanged following fire. In general, changes in soil properties following fire were greatest at the 0-5 cm layer and more modest at the 5-10 cm sampling depth. These changes were related primarily to oxidation of the detrital layer during fire and concurrent changes in the soil environment

  1. The relationship of normal body temperature, end-expired breath temperature, and BAC/BrAC ratio in 98 physically fit human test subjects.

    Science.gov (United States)

    Cowan, J Mack; Burris, James M; Hughes, James R; Cunningham, Margaret P

    2010-06-01

    The relationship between normal body temperature, end-expired breath temperature, and blood alcohol concentration (BAC)/breath alcohol concentration (BrAC) ratio was studied in 98 subjects (84 men, 14 women). Subjects consumed alcohol sufficient to produce a BrAC of at least 0.06 g/210 L 45-75 min after drinking. Breath samples were analyzed using an Intoxilyzer 8000 specially equipped to measure breath temperature. Venous blood samples and body temperatures were then taken. The mean body temperature of the men (36.6 degrees C) was lower than the women (37.0 degrees C); however, their mean breath temperatures were virtually identical (men: 34.5 degrees C; women: 34.6 degrees C). The BAC exceeded the BrAC for every subject. BAC/BrAC ratios were calculated from the BAC and BrAC analytical results. There was no difference in the BAC/BrAC ratios for men (1:2379) and women (1:2385). The correlation between BAC and BrAC was high (r = 0.938, p body temperature and end-expired breath temperature, body temperature and BAC/BrAC ratio, and breath temperature and BAC/BrAC ratio were much lower. Neither normal body temperature nor end-expired breath temperature was strongly associated with BAC/BrAC ratio.

  2. Desarrollo de un modelo de evaluación financiera de proyectos para BAC Credomatic Network

    OpenAIRE

    Sandí Piedra, Allan Adolfo

    2016-01-01

    Universidad de Costa Rica. Posgrado en Administración y Dirección de Empresas. Maestría Profesional en Administración y Dirección de Empresas con énfasis en Finanzas, 2016 El presente trabajo final de investigación contiene los aspectos que fundamentan un modelo de Evaluación Financiera de Proyectos que permite solventar la problemática identificada en BAC Credomatic Network respecto al limitado valor agregado del proceso existente de valoración financiera de los proyectos de la empresa. P...

  3. Toward forward genetic screens in malaria-causing parasites using the piggyBac transposon

    Directory of Open Access Journals (Sweden)

    de Koning-Ward Tania F

    2011-03-01

    Full Text Available Abstract The ability to analyze gene function in malaria-causing Plasmodium parasites has received a boost with a recent paper in BMC Genomics that describes a genome-wide mutagenesis system in the rodent malaria species Plasmodium berghei using the transposon piggyBac. This advance holds promise for identifying and validating new targets for intervention against malaria. But further improvements are still needed for the full power of genome-wide molecular genetic screens to be utilized in this organism. See research article: http://www.biomedcentral.com/1471-2164/12/155

  4. Expression of Human CAR Splicing Variants in BAC-Transgenic Mice

    OpenAIRE

    Zhang, Yu-Kun Jennifer; Lu, Hong; Klaassen, Curtis D.

    2012-01-01

    The nuclear receptor constitutive androstane receptor (CAR) is a key regulator for drug metabolism in liver. Human CAR (hCAR) transcripts are subjected to alternative splicing. Some hCAR splicing variants (SVs) have been shown to encode functional proteins by reporter assays. However, in vivo research on the activity of these hCAR SVs has been impeded by the absence of a valid model. This study engineered an hCAR-BAC-transgenic (hCAR-TG) mouse model by integrating the 8.5-kbp hCAR gene as wel...

  5. Characterization of pig farms in Hung Yen, Hai Duong and Bac Ninh provinces

    OpenAIRE

    Nguyen Van Duy; Vu Dinh; Dang Vu, Binh; Vo Trong, Thanh; Nguyen Cong, oanh; Phan Van, Chung

    2007-01-01

    In order to characterization of pig farms in the Red River Delta, a study was conducted on 90 pig farms in Hung Yen, Hai Duong and Bac Ninh provinces from June to December 2006. Results show that most of the pig farms had been built for five years with a small size (0.5 hectare per farm). The invested capital was about 300-400 millions VND per farm. Four main sow groups used in the farms included crossbred exotic sows (51.1%), crossbred sow between local and exotic breeds (14.4%), purebred La...

  6. BACS: The Brussels Artificial Character Sets for studies in cognitive psychology and neuroscience.

    Science.gov (United States)

    Vidal, Camille; Content, Alain; Chetail, Fabienne

    2017-12-01

    Written symbols such as letters have been used extensively in cognitive psychology, whether to understand their contributions to written word recognition or to examine the processes involved in other mental functions. Sometimes, however, researchers want to manipulate letters while removing their associated characteristics. A powerful solution to do so is to use new characters, devised to be highly similar to letters, but without the associated sound or name. Given the growing use of artificial characters in experimental paradigms, the aim of the present study was to make available the Brussels Artificial Character Sets (BACS): two full, strictly controlled, and portable sets of artificial characters for a broad range of experimental situations.

  7. Tolerancia experimental de las especies vegetales Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea al boro, en Argentina

    Directory of Open Access Journals (Sweden)

    Marta L. de Viana

    2008-09-01

    Full Text Available La actividad de las industrias borateras constituye una fuente puntual y difusa de contaminación del aire, suelo y aguas superficiales y profundas. Por lo tanto, el estudio y experimentación acerca de las posibles formas de contrarrestar este impacto constituye una prioridad. Una técnica relativamente nueva para descontaminar suelos es la fitorremediación, que emplea plantas y microorganismos asociados. El primer paso es detectar las especies vegetales tolerantes, lo que constituye el objetivo de este trabajo. Se realizó un experimento en laboratorio para evaluar la germinación, la supervivencia y el crecimiento de distintas especies en diferentes concentraciones de boro. Al comienzo y al final del experimento se determinó la concentración de boro en el sustrato para cada tratamiento y para sustratos con y sin vegetación. Se encontraron diferencias significativas debidas al tratamiento, la especie y la interacción especie *tratamiento. M. sativa, N. glauca y J. mimosifolia fueron las especies de mayor tolerancia al boro. Las otras especies presentaron una disminución en todas las variables-respuesta en función de la concentración del contaminante. Todas presentaron una baja supervivencia en la máxima concentración. La disminución de boro fue máxima en el tratamiento de 30 ppm de boro con M. sativa y la menor se registró en los tratamiento de 20 ppm de boro con J. mimosifolia y de 30 ppm de boro con T. stans y S. oleraceae. Se concluye que N. glauca, M. sativa y J. mimosifolia podrían considerarse como prometedoras en remediación.Experimental tolerance to boron of the plant species Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea in Argentina. The activity of boron industries is a punctual and diffuse source of air, soil and water pollution. Therefore, it is a priority to study possible ways of reducing this impact. A relatively new technology for reducing soil pollution is

  8. MacroBac: New Technologies for Robust and Efficient Large-Scale Production of Recombinant Multiprotein Complexes.

    Science.gov (United States)

    Gradia, Scott D; Ishida, Justin P; Tsai, Miaw-Sheue; Jeans, Chris; Tainer, John A; Fuss, Jill O

    2017-01-01

    Recombinant expression of large, multiprotein complexes is essential and often rate limiting for determining structural, biophysical, and biochemical properties of DNA repair, replication, transcription, and other key cellular processes. Baculovirus-infected insect cell expression systems are especially well suited for producing large, human proteins recombinantly, and multigene baculovirus systems have facilitated studies of multiprotein complexes. In this chapter, we describe a multigene baculovirus system called MacroBac that uses a Biobricks-type assembly method based on restriction and ligation (Series 11) or ligation-independent cloning (Series 438). MacroBac cloning and assembly is efficient and equally well suited for either single subcloning reactions or high-throughput cloning using 96-well plates and liquid handling robotics. MacroBac vectors are polypromoter with each gene flanked by a strong polyhedrin promoter and an SV40 poly(A) termination signal that minimize gene order expression level effects seen in many polycistronic assemblies. Large assemblies are robustly achievable, and we have successfully assembled as many as 10 genes into a single MacroBac vector. Importantly, we have observed significant increases in expression levels and quality of large, multiprotein complexes using a single, multigene, polypromoter virus rather than coinfection with multiple, single-gene viruses. Given the importance of characterizing functional complexes, we believe that MacroBac provides a critical enabling technology that may change the way that structural, biophysical, and biochemical research is done. © 2017 Elsevier Inc. All rights reserved.

  9. Construction of an Americn mink Bacterial Artificial Chromosome (BAC) library and sequencing candidate genes important for the fur industry

    DEFF Research Database (Denmark)

    Anistoroaei, Razvan Marian; Hallers, Boudewijn ten; Nefedov, Michael

    2011-01-01

    BACKGROUND: Bacterial artificial chromosome (BAC) libraries continue to be invaluable tools for the genomic analysis of complex organisms. Complemented by the newly and fast growing deep sequencing technologies, they provide an excellent source of information in genomics projects. RESULTS: Here, we...... report the construction and characterization of the CHORI-231 BAC library constructed from a Danish-farmed, male American mink (Neovison vison). The library contains approximately 165,888 clones with an average insert size of 170 kb, representing approximately 10-fold coverage. High-density filters, each...... consisting of 18,432 clones spotted in duplicate, have been produced for hybridization screening and are publicly available. Overgo probes derived from expressed sequence tags (ESTs), representing 21 candidate genes for traits important for the mink industry, were used to screen the BAC library...

  10. Effect of mercaptoethylamine on DNA degradation in thermophilic bacteria Bac. stearothermophilus exposed to. gamma. -, UV-radiation or methylnitrosourea

    Energy Technology Data Exchange (ETDEWEB)

    Fomenko, L A; Kuznetsovea, E A; Gaziev, A I

    1984-07-01

    The effect of mercaptoethylamine (MEA) on degradation of DNA in thermophilic bacteria Bac. stear. exposed to ..gamma..-, UV-rays or methylnitrosourea (MNU) was studied. Using centrifugation on alkaline and neutral sucrose gradients, it was shown that MEA inhibits the accumulation of breaks in the DNA of Bac. stear. It also lowers the level of DNA degradation in toluene-treated cells of Bac. stear. under the action of the intrinsic nuclease, reduces the activity of the endonuclease specific for apurinic DNA, as well as that of S/sub 1/-nuclease and DNase-I in vitro. The inhibition in the accumulation of DNA breaks is assumed to be due to a decrease of the endonuclease activity in the cells of thermophilic bacteria.

  11. Effect of mercaptoethylamine on DNA degradation in thermophilic bacteria Bac. stearothermophilus exposed to γ-, UV-radiation or methylnitrosourea

    International Nuclear Information System (INIS)

    Fomenko, L.A.; Kuznetsovea, E.A.; Gaziev, A.I.

    1984-01-01

    The effect of mercaptoethylamine (MEA) on degradation of DNA in thermophilic bacteria Bac. stear. exposed to γ-, UV-rays or methylnitrosourea (MNU) was studied. Using centrifugation on alkaline and neutral sucrose gradients, it was shown that MEA inhibits the accumulation of breaks in the DNA of Bac. stear. It also lowers the level of DNA degradation in toluene-treated cells of Bac. stear. under the action of the intrinsic nuclease, reduces the activity of the endonuclease specific for apurinic DNA, as well as that of S 1 -nuclease and DNase-I in vitro. The inhibition in the accumulation of DNA breaks is assumed to be due to a decrease of the endonuclease activity in the cells of thermophilic bacteria. (orig.)

  12. MBBR evaluation for oil refinery wastewater treatment, with post-ozonation and BAC, for wastewater reuse.

    Science.gov (United States)

    Schneider, E E; Cerqueira, A C F P; Dezotti, M

    2011-01-01

    This work evaluated the performance of a Moving Bed Biofilm Reactor (MBBR) in the treatment of an oil refinery wastewater. Also, it investigated the possibility of reuse of the MBBR effluent, after ozonation in series with a biological activated carbon (BAC) column. The best performance of the MBBR was achieved with a hydraulic retention time (HRT) of 6 hours, employing a bed to bioreactor volume ratio (V(B)/V(R)) of 0.6. COD and N-NH₄(+) MBBR effluent concentrations ranged from 40 to 75 mg L⁻¹ (removal efficiency of 69-89%) and 2 to 6 mg L⁻¹ (removal efficiency of 45-86%), respectively. Ozonation carried out for 15 min with an ozone concentration of 5 mg L⁻¹ was able to improve the treated wastewater biodegradability. The treatment performance of the BAC columns was practically the same for ozonated and non ozonated MBBR effluents. The dissolved organic carbon (DOC) content of the columns of the activated carbon columns (CAG) was in the range of 2.1-3.8 mg L⁻¹, and the corresponding DOC removal efficiencies were comprised between 52 and 75%. The effluent obtained at the end of the proposed treatment presented a quality, which meet the requirements for water reuse in the oil refinery.

  13. A new normalizing algorithm for BAC CGH arrays with quality control metrics.

    Science.gov (United States)

    Miecznikowski, Jeffrey C; Gaile, Daniel P; Liu, Song; Shepherd, Lori; Nowak, Norma

    2011-01-01

    The main focus in pin-tip (or print-tip) microarray analysis is determining which probes, genes, or oligonucleotides are differentially expressed. Specifically in array comparative genomic hybridization (aCGH) experiments, researchers search for chromosomal imbalances in the genome. To model this data, scientists apply statistical methods to the structure of the experiment and assume that the data consist of the signal plus random noise. In this paper we propose "SmoothArray", a new method to preprocess comparative genomic hybridization (CGH) bacterial artificial chromosome (BAC) arrays and we show the effects on a cancer dataset. As part of our R software package "aCGHplus," this freely available algorithm removes the variation due to the intensity effects, pin/print-tip, the spatial location on the microarray chip, and the relative location from the well plate. removal of this variation improves the downstream analysis and subsequent inferences made on the data. Further, we present measures to evaluate the quality of the dataset according to the arrayer pins, 384-well plates, plate rows, and plate columns. We compare our method against competing methods using several metrics to measure the biological signal. With this novel normalization algorithm and quality control measures, the user can improve their inferences on datasets and pinpoint problems that may arise in their BAC aCGH technology.

  14. MUTATION ON Bacillus subtilis BAC4 USING ACRIDINE ORANGE AS AN EFFORT FOR INCREASING ANTIBIOTIC PRODUCTION

    Directory of Open Access Journals (Sweden)

    Supartono Supartono

    2010-06-01

    Full Text Available The efforts to get a new antibiotic require to be done continuously, because infection diseases still become the main health problems in Indonesia. A new local strain of Bacillus subtilis BAC4 has been known producing an antibiotic that inhibites Serratia marcescens ATCC 27117 growth. Nevertheless, the optimum conditions have not been studied seriously. The objective of this research was to conduct mutation on B. subtilis BAC4 in order to obtain a mutant cell that overproduct in producing antibiotic. The mutation process was performed by using acridine orange of 1 g.L-1 randomly at various volumes. The production of antibiotic was conducted using batch fermentation and antibiotic assay was performed with agar absorption method using S.  marcescens ATCC 27117 as bacteria assay. Research result provided a B. subtilis M10 mutant with overproduction of antibiotic. Characterization of B. subtilis M10 mutant showed that the mutant cell has size of (0.5-1.0 µm x (1.85-2.5 µm; spore has the form of ellipse with thick wavy wall, positive reaction for catalase, and forming acid from glucose and xylose.   Keywords: mutant, Bacillus, acridin, and antibiotics

  15. BacHbpred: Support Vector Machine Methods for the Prediction of Bacterial Hemoglobin-Like Proteins

    Directory of Open Access Journals (Sweden)

    MuthuKrishnan Selvaraj

    2016-01-01

    Full Text Available The recent upsurge in microbial genome data has revealed that hemoglobin-like (HbL proteins may be widely distributed among bacteria and that some organisms may carry more than one HbL encoding gene. However, the discovery of HbL proteins has been limited to a small number of bacteria only. This study describes the prediction of HbL proteins and their domain classification using a machine learning approach. Support vector machine (SVM models were developed for predicting HbL proteins based upon amino acid composition (AC, dipeptide composition (DC, hybrid method (AC + DC, and position specific scoring matrix (PSSM. In addition, we introduce for the first time a new prediction method based on max to min amino acid residue (MM profiles. The average accuracy, standard deviation (SD, false positive rate (FPR, confusion matrix, and receiver operating characteristic (ROC were analyzed. We also compared the performance of our proposed models in homology detection databases. The performance of the different approaches was estimated using fivefold cross-validation techniques. Prediction accuracy was further investigated through confusion matrix and ROC curve analysis. All experimental results indicate that the proposed BacHbpred can be a perspective predictor for determination of HbL related proteins. BacHbpred, a web tool, has been developed for HbL prediction.

  16. Cytogenetical anchoring of sheep linkage map and syntenic groups using a sheep BAC library

    Directory of Open Access Journals (Sweden)

    Cribiu Edmond-Paul

    2000-07-01

    Full Text Available Abstract In order to simultaneously integrate linkage and syntenic groups to the ovine chromosomal map, a sheep bacterial artificial chromosome (BAC library was screened with previously assigned microsatellites using a sheep-hamster hybrid panel and genetic linkage. Thirty-three BACs were obtained, fluorescently labelled and hybridised on sheep-goat hybrid metaphases (2n = 57. This study allowed us, (i, to anchor all linkage groups on sheep chromosomes, (ii, to give information on the probable position of the centromere on the linkage map for the centromeric chromosomes, (iii, to contradict the previous orientation of the ovine × linkage group by the mapping of BMS1008 on OARXq38. Concerning our somatic cell hybrid panel, this study resulted in the assignment of all the previously unassigned groups to ovine chromosomes and a complete characterisation of the hybrid panel. In addition, since hybridisations were performed on a sheep-goat hybrid, new marker/anchoring points were added to the caprine cytogenetic map.

  17. A New Normalizing Algorithm for BAC CGH Arrays with Quality Control Metrics

    Directory of Open Access Journals (Sweden)

    Jeffrey C. Miecznikowski

    2011-01-01

    Full Text Available The main focus in pin-tip (or print-tip microarray analysis is determining which probes, genes, or oligonucleotides are differentially expressed. Specifically in array comparative genomic hybridization (aCGH experiments, researchers search for chromosomal imbalances in the genome. To model this data, scientists apply statistical methods to the structure of the experiment and assume that the data consist of the signal plus random noise. In this paper we propose “SmoothArray”, a new method to preprocess comparative genomic hybridization (CGH bacterial artificial chromosome (BAC arrays and we show the effects on a cancer dataset. As part of our R software package “aCGHplus,” this freely available algorithm removes the variation due to the intensity effects, pin/print-tip, the spatial location on the microarray chip, and the relative location from the well plate. removal of this variation improves the downstream analysis and subsequent inferences made on the data. Further, we present measures to evaluate the quality of the dataset according to the arrayer pins, 384-well plates, plate rows, and plate columns. We compare our method against competing methods using several metrics to measure the biological signal. With this novel normalization algorithm and quality control measures, the user can improve their inferences on datasets and pinpoint problems that may arise in their BAC aCGH technology.

  18. Genome-wide target profiling of piggyBac and Tol2 in HEK 293: pros and cons for gene discovery and gene therapy

    Science.gov (United States)

    2011-01-01

    Background DNA transposons have emerged as indispensible tools for manipulating vertebrate genomes with applications ranging from insertional mutagenesis and transgenesis to gene therapy. To fully explore the potential of two highly active DNA transposons, piggyBac and Tol2, as mammalian genetic tools, we have conducted a side-by-side comparison of the two transposon systems in the same setting to evaluate their advantages and disadvantages for use in gene therapy and gene discovery. Results We have observed that (1) the Tol2 transposase (but not piggyBac) is highly sensitive to molecular engineering; (2) the piggyBac donor with only the 40 bp 3'-and 67 bp 5'-terminal repeat domain is sufficient for effective transposition; and (3) a small amount of piggyBac transposases results in robust transposition suggesting the piggyBac transpospase is highly active. Performing genome-wide target profiling on data sets obtained by retrieving chromosomal targeting sequences from individual clones, we have identified several piggyBac and Tol2 hotspots and observed that (4) piggyBac and Tol2 display a clear difference in targeting preferences in the human genome. Finally, we have observed that (5) only sites with a particular sequence context can be targeted by either piggyBac or Tol2. Conclusions The non-overlapping targeting preference of piggyBac and Tol2 makes them complementary research tools for manipulating mammalian genomes. PiggyBac is the most promising transposon-based vector system for achieving site-specific targeting of therapeutic genes due to the flexibility of its transposase for being molecularly engineered. Insights from this study will provide a basis for engineering piggyBac transposases to achieve site-specific therapeutic gene targeting. PMID:21447194

  19. Application of EDTA-functionalized bamboo activated carbon (BAC) for Pb(II) and Cu(II) removal from aqueous solutions

    Science.gov (United States)

    Lv, Dan; Liu, Yu; Zhou, Jiasheng; Yang, Kunlun; Lou, Zimo; Baig, Shams Ali; Xu, Xinhua

    2018-01-01

    In this study, a novel bamboo activated carbon (BAC) with ethylene diamine tetraacetic acid (EDTA) functionality was prepared by direct grafting in the presence of tetraethyl orthosilicate (TEOS) as a crosslinking agent. The BAC@SiO2-EDTA was characterized by SEM, TEM, TGA, FTIR, XPS and its adsorption property for removal of Pb(II) and Cu(II) under various experimental conditions was also investigated. The characterization results reflected that EDTA was successfully assembled on the surface of the BAC and average pore size increased from 4.10 to 4.83 nm as BAC grafted with EDTA. Adsorption data fitted very well in Langmuir isotherm model and pseudo-second-order kinetic model. As compared with the raw BAC, the maximum adsorption capacities of BAC@SiO2-EDTA for the Pb(II) and Cu(II) increased from 45.45 to 123.45 mg g-1 and from 6.85 to 42.19 mg g-1, since the existence of EDTA on modified BAC promoted the formation of chemical complex. The removal of heavy metal ions mainly depended on the complexation with EDTA and the electrostatic attractions with negatively charged surface of BAC@SiO2-EDTA. The adsorption of Pb(II)/Cu(II) on the BAC@SiO2-EDTA was pH dependent and pH 5-6 was considered an optimum. However, lower temperature favored the adsorption and the maximum adsorption was recorded at 20 °C. In addition, BAC@SiO2-EDTA had an excellent reusability with about 40% decline in the adsorption capacity for Pb(II) after fifth reuse. Insignificant influences of co-existing cations and natural organic matter (NOM) were found on the adsorption of Pb(II) and Cu(II). All the results demonstrate that BAC@SiO2-EDTA is a potential adsorbent for metal ions in wastewater.

  20. Construction of a nurse shark (Ginglymostoma cirratum) bacterial artificial chromosome (BAC) library and a preliminary genome survey.

    Science.gov (United States)

    Luo, Meizhong; Kim, Hyeran; Kudrna, Dave; Sisneros, Nicholas B; Lee, So-Jeong; Mueller, Christopher; Collura, Kristi; Zuccolo, Andrea; Buckingham, E Bryan; Grim, Suzanne M; Yanagiya, Kazuyo; Inoko, Hidetoshi; Shiina, Takashi; Flajnik, Martin F; Wing, Rod A; Ohta, Yuko

    2006-05-03

    Sharks are members of the taxonomic class Chondrichthyes, the oldest living jawed vertebrates. Genomic studies of this group, in comparison to representative species in other vertebrate taxa, will allow us to theorize about the fundamental genetic, developmental, and functional characteristics in the common ancestor of all jawed vertebrates. In order to obtain mapping and sequencing data for comparative genomics, we constructed a bacterial artificial chromosome (BAC) library for the nurse shark, Ginglymostoma cirratum. The BAC library consists of 313,344 clones with an average insert size of 144 kb, covering ~4.5 x 1010 bp and thus providing an 11-fold coverage of the haploid genome. BAC end sequence analyses revealed, in addition to LINEs and SINEs commonly found in other animal and plant genomes, two new groups of nurse shark-specific repetitive elements, NSRE1 and NSRE2 that seem to be major components of the nurse shark genome. Screening the library with single-copy or multi-copy gene probes showed 6-28 primary positive clones per probe of which 50-90% were true positives, demonstrating that the BAC library is representative of the different regions of the nurse shark genome. Furthermore, some BAC clones contained multiple genes, making physical mapping feasible. We have constructed a deep-coverage, high-quality, large insert, and publicly available BAC library for a cartilaginous fish. It will be very useful to the scientific community interested in shark genomic structure, comparative genomics, and functional studies. We found two new groups of repetitive elements specific to the nurse shark genome, which may contribute to the architecture and evolution of the nurse shark genome.

  1. Construction of a nurse shark (Ginglymostoma cirratum bacterial artificial chromosome (BAC library and a preliminary genome survey

    Directory of Open Access Journals (Sweden)

    Inoko Hidetoshi

    2006-05-01

    Full Text Available Abstract Background Sharks are members of the taxonomic class Chondrichthyes, the oldest living jawed vertebrates. Genomic studies of this group, in comparison to representative species in other vertebrate taxa, will allow us to theorize about the fundamental genetic, developmental, and functional characteristics in the common ancestor of all jawed vertebrates. Aims In order to obtain mapping and sequencing data for comparative genomics, we constructed a bacterial artificial chromosome (BAC library for the nurse shark, Ginglymostoma cirratum. Results The BAC library consists of 313,344 clones with an average insert size of 144 kb, covering ~4.5 × 1010 bp and thus providing an 11-fold coverage of the haploid genome. BAC end sequence analyses revealed, in addition to LINEs and SINEs commonly found in other animal and plant genomes, two new groups of nurse shark-specific repetitive elements, NSRE1 and NSRE2 that seem to be major components of the nurse shark genome. Screening the library with single-copy or multi-copy gene probes showed 6–28 primary positive clones per probe of which 50–90% were true positives, demonstrating that the BAC library is representative of the different regions of the nurse shark genome. Furthermore, some BAC clones contained multiple genes, making physical mapping feasible. Conclusion We have constructed a deep-coverage, high-quality, large insert, and publicly available BAC library for a cartilaginous fish. It will be very useful to the scientific community interested in shark genomic structure, comparative genomics, and functional studies. We found two new groups of repetitive elements specific to the nurse shark genome, which may contribute to the architecture and evolution of the nurse shark genome.

  2. Aerobic biodegradation of a sulfonated phenylazonaphthol dye by a bacterial community immobilized in a multistage packed-bed BAC reactor.

    Science.gov (United States)

    Ruiz-Arias, Alfredo; Juárez-Ramírez, Cleotilde; de los Cobos-Vasconcelos, Daniel; Ruiz-Ordaz, Nora; Salmerón-Alcocer, Angélica; Ahuatzi-Chacón, Deifilia; Galíndez-Mayer, Juvencio

    2010-11-01

    A microbial community able to aerobically degrade the azo dye Acid Orange 7 was selected from riparian or lacustrine sediments collected at sites receiving textile wastewaters. Three bacterial strains, pertaining to the genera Pseudomonas, Arthrobacter, and Rhizobium, constitute the selected community. The biodegradation of AO7 was carried out in batch-suspended cell culture and in a continuously operated multistage packed-bed BAC reactor. The rapid decolorization observed in batch culture, joined to a delay of about 24 h in COD removal and cell growth, suggests that enzymes involved in biodegradation of the aromatic amines generated after AO7 azo-bond cleavage (1-amino-2-naphthol [1-A2N] and 4-aminobenzenesulfonic acid [4-ABS]), are inducible in this microbial consortium. After this presumptive induction period, the accumulated byproducts, measured through COD, were partially metabolized and transformed in cell mass. At all azo dye loading rates used, complete removal of AO7 and 1-A2N was obtained in the multistage packed-bed BAC reactor (PBR).; however, the overall COD (eta ( COD )) and 4-ABS (eta ( ABS )) removal efficiencies obtained in steady state continuous culture were about 90%. Considering the toxicity of 1-A2N, its complete removal has particular relevance. In the first stages of the packed-bed BAC reactor (Fig. 4a-c), major removal was observed. In the last stage, only a slight removal of COD and 4-ABS was obtained. Comparing to several reported studies, the continuously operated multistage packed-bed BAC reactor showed similar or superior results. In addition, the operation of large-packed-bed BAC reactors could be improved by using several shallow BAC bed stages, because the pressure drop caused by bed compaction of a support material constituted by small and fragile particles can be reduced.

  3. Genetic variation and control of chloroplast pigment concentrations in Picea rubens, Picea mariana and their hybrids. I. Ambient and elevated [CO2] environments

    International Nuclear Information System (INIS)

    Major, J.E.; Barsi, D.C.; Mosseler, A.; Campbell, M.

    2007-01-01

    A significant decline has been noted in the red spruce component of the Acadian forest region in eastern Canada and the northeastern United States as a result of excessive harvesting, acid rain, and global warming. Two experiments were performed to acquire benchmark adaptive traits for information from a red spruce (RS) (Picea rubens Sargand) and black spruce (BS) (P. mariana (Mill.) B.S.P.) genetic complex grown in ambient carbon dioxide concentration ([CO 2 ]). The first experiment involved RS-BS seed sources from across the RS geographical range, while the second experiment involved an intra- and interspecific controlled-cross experiment to determine if RS and BS have unique chloroplast pigment concentrations and traits that reflect adaptations to different ecological niches. The objective was to determine species origin and hybrid variations in chloroplast pigment concentrations; examine the effect of elevated [CO 2 ] on chloroplast pigments; determine the inheritance of chloroplast pigments and examine the relationship of chloroplast pigment concentrations of trees grown at ambient [CO 2 ] with productivity traits and nitrogen concentrations. The traits related to light-energy processing have pronounced ecological implications for plant health. Results from the species origin experiment showed that total chlorophyll concentration was about 15 per cent higher in ambient [CO 2 ] than in elevated [CO 2 ]. In ambient [CO 2 ], BS populations had 11 per cent higher total chlorophyll and carotenoid concentrations than RS populations. Results from the controlled-cross experiment showed that families with a hybrid index of 25 per cent RS had the highest total chlorophyll concentrations, and families with hybrid indices of 75 and 100 had the lowest amounts. A predominant male effect for chlorophyll concentration was noted. In ambient and elevated [CO 2 ] environments, crosses with BS males had 10.6 and 17.6 per cent higher total chlorophyll concentrations than crosses

  4. Regulation of somatic embryo development in Norway spruce (Picea abies). A molecular approach to the characterization of specific developmental stages

    Energy Technology Data Exchange (ETDEWEB)

    Sabala, I. [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Forest Genetics

    1998-12-31

    Embryo development is a complex process involving a set of strictly regulated events. The regulation of these events is poorly understood especially during the early stages of embryo development. Somatic embryos go through the same developmental stages as zygotic embryos making them an ideal model system for studying the regulation of embryo development. We have used embryogenic cultures of Picea abies to study some aspects of the regulation of embryo development in gymnosperms. The bottle neck during somatic embryogenesis is the switch from the proliferation stage to the maturation stage. This switch is initiated by giving somatic embryos a maturation treatment i.e. the embryos are treated with abscisic acid (ABA). Somatic embryos which respond to ABA by forming mature somatic embryos were stimulated to secret a 70 kDa protein, AF70. The af70 gene was isolated and characterised. The expression of the af70 gene was constitutive in embryos but was highly ABA-induced in seedlings. Moreover, expression of this gene was stimulated during cold acclimation of Picea abies seedlings. A full length Picea abies cDNA clone Pa18, encoding a protein with the characteristics of plant lipid transfer proteins (LTPs), was isolated and characterised. The Pa18 gene is constitutively expressed in embryogenic cultures of Picea abies representing different stages of development as well as in nonembryogenic callus and seedlings. In situ hybridization showed that Pa18 gene is expressed in all embryonic cells of proliferating somatic embryos but the expression of the gene in mature somatic and zygotic embryos is restricted to the outer cell layer. Southern blot analysis at different stringencies was consistent with a single gene. An alteration in expression of Pa18 causes disturbance in the formation of the proper outer cell layer in the maturing somatic embryos. In addition to its influence on embryo development the Pa18 gene product also inhibits growth of Agrobacterium tumefaciens 195

  5. Long-term Responses of Canopy-understorey Interactions to Disturbance Severity in Primary Picea abies Forests

    Czech Academy of Sciences Publication Activity Database

    Bače, R.; Schurman, J.S.; Brabec, Marek; Čada, V.; Deprés, T.; Janda, P.; Lábusová, J.; Mikoláš, M.; Morrissey, R. C.; Mrhalová, H.; Nagel, T.A.; Nováková, M. H.; Seedre, M.; Synek, M.; Trotsiuk, V.; Svoboda, M.

    2017-01-01

    Roč. 28, č. 6 (2017), s. 1128-1139 ISSN 1100-9233 Grant - others:GA ČR(CZ) GA15-14840S Institutional support: RVO:67985807 Keywords : Disturbance regime * Natural regeneration * Primary forest * Picea abies (L.) Karst * Windstorms * Bark beetle * Understory light availability * Saplings and poles * Canopy openness * Mountain forest Subject RIV: BB - Applied Statistics, Operational Research OBOR OECD: Statistics and probability Impact factor: 2.924, year: 2016

  6. Seasonal changes of Rubisco content and activity in Fagus sylvatica and Picea abies affected by elevated CO2 concentration

    Czech Academy of Sciences Publication Activity Database

    Hrstka, M.; Urban, Otmar; Babák, L.

    2012-01-01

    Roč. 66, č. 9 (2012), s. 836-841 ISSN 0366-6352 R&D Projects: GA AV ČR IAA600870701; GA MŠk(CZ) LM2010007; GA MŠk(CZ) ED1.1.00/02.0073 Institutional research plan: CEZ:AV0Z60870520 Keywords : Rubisco content * Rubisco activity * seasonal changes * elevated CO2 concentrations * Fagus sylvatica * Picea abies Subject RIV: EH - Ecology, Behaviour Impact factor: 0.879, year: 2012

  7. The relationship between species diversity and genetic structure in the rare Picea chihuahuana tree species community, Mexico.

    Science.gov (United States)

    Simental-Rodríguez, Sergio Leonel; Quiñones-Pérez, Carmen Zulema; Moya, Daniel; Hernández-Tecles, Enrique; López-Sánchez, Carlos Antonio; Wehenkel, Christian

    2014-01-01

    Species diversity and genetic diversity, the most basic elements of biodiversity, have long been treated as separate topics, although populations evolve within a community context. Recent studies on community genetics and ecology have suggested that genetic diversity is not completely independent of species diversity. The Mexican Picea chihuahuana Martínez is an endemic species listed as "Endangered" on the Red List. Forty populations of Chihuahua spruce have been identified. This species is often associated with tree species of eight genera in gallery forests. This rare Picea chihuahuana tree community covers an area no more than 300 ha and has been subject of several studies involving different topics such as ecology, genetic structure and climate change. The overall aim of these studies was to obtain a dataset for developing management tools to help decision makers implement preservation and conservation strategies. However, this unique forest tree community may also represent an excellent subject for helping us to understand the interplay between ecological and evolutionary processes in determining community structure and dynamics. The AFLP technique and species composition data were used together to test the hypothesis that species diversity is related to the adaptive genetic structure of some dominant tree species (Picea chihuahuana, Pinus strobiformis, Pseudotsuga menziesii and Populus tremuloides) of the Picea chihuahuana tree community at fourteen locations. The Hill numbers were used as a diversity measure. The results revealed a significant correlation between tree species diversity and genetic structure in Populus tremuloides. Because the relationship between the two levels of diversity was found to be positive for the putative adaptive AFLP detected, genetic and species structures of the tree community were possibly simultaneously adapted to a combination of ecological or environmental factors. The present findings indicate that interactions between

  8. Effects of aluminum on growth, polyamine metabolism, and inorganic ions in suspension cultures of red spruce (Picea rubens)

    Science.gov (United States)

    Rakesh Minocha; Walter C. Shortle; Daniel J. Jr. Coughin; Subhash C. Minocha

    1996-01-01

    The influence of age of red spruce (Picea rubens Sarg.) cell suspensions on aluminum (Al) effects was studied by adding AICI3 (0.2, 0.5, and 1.0 mM) to the media on each day of a 7-day culture period and analyzing for changes in total cell mass, polyamines, arginine decarboxylase activity, and inorganic ions after 24 h of...

  9. Biomass equations and biomass expansion factors (BEFs) for pine (pinus spp.), spruce (picea spp.) and broadleaved dominated stands in Norway

    OpenAIRE

    Viken, Knut Ole

    2012-01-01

    Abstract The objectives of this study were (1) to develop models for estimation of stand-level tree biomass for spruce (picea spp.)- pine (pinus spp.)- and broadleaved-dominated forest in Norway and, (2) develop biomass expansion factors (BEFs; ratio of stem volume to biomass) which convert stem volume to whole tree biomass for Norwegian forest conditions. A dataset from a 5 year period (2006 – 2010) from the Norwegian National Forest Inventory (NFI) were used to develop the...

  10. DNA-Methylierung nach abiotischen und biotischen Einflüssen und Expressionsanalyse pathogeninduzierter Gene in Picea abies (L.) Karst.

    OpenAIRE

    Baumann, Ruediger

    2006-01-01

    Basierend auf Beobachtungen über phänotypische Änderungen in Vollgeschwisterfamilien der Fichte,Picea abies, aus unterschiedlichen Kreuzungsumwelten wurde nach der geneti-schen Untersuchung mit EST-Markern eine kapillarelektrophoretische Untersuchung der DNA durchgeführt. Dabei wurden signifikante Unterschiede im Methylcytosin(mC)-Gehalt festgestellt. Messungen der mC-Gehalte nach Hitzeschock zeigten, den Einfluss der Um-weltbedingungen auf die Methylierung der DNA. Untersuchungen zur Dynamik...

  11. Giant panda BAC library construction and assembly of a 650-kb contig spanning major histocompatibility complex class II region

    Directory of Open Access Journals (Sweden)

    Pan Hui-Juan

    2007-09-01

    Full Text Available Abstract Background Giant panda is rare and endangered species endemic to China. The low rates of reproductive success and infectious disease resistance have severely hampered the development of captive and wild populations of the giant panda. The major histocompatibility complex (MHC plays important roles in immune response and reproductive system such as mate choice and mother-fetus bio-compatibility. It is thus essential to understand genetic details of the giant panda MHC. Construction of a bacterial artificial chromosome (BAC library will provide a new tool for panda genome physical mapping and thus facilitate understanding of panda MHC genes. Results A giant panda BAC library consisting of 205,800 clones has been constructed. The average insert size was calculated to be 97 kb based on the examination of 174 randomly selected clones, indicating that the giant panda library contained 6.8-fold genome equivalents. Screening of the library with 16 giant panda PCR primer pairs revealed 6.4 positive clones per locus, in good agreement with an expected 6.8-fold genomic coverage of the library. Based on this BAC library, we constructed a contig map of the giant panda MHC class II region from BTNL2 to DAXX spanning about 650 kb by a three-step method: (1 PCR-based screening of the BAC library with primers from homologous MHC class II gene loci, end sequences and BAC clone shotgun sequences, (2 DNA sequencing validation of positive clones, and (3 restriction digest fingerprinting verification of inter-clone overlapping. Conclusion The identifications of genes and genomic regions of interest are greatly favored by the availability of this giant panda BAC library. The giant panda BAC library thus provides a useful platform for physical mapping, genome sequencing or complex analysis of targeted genomic regions. The 650 kb sequence-ready BAC contig map of the giant panda MHC class II region from BTNL2 to DAXX, verified by the three-step method, offers a

  12. Brucella detection in blood: comparison of the BacT/Alert standard aerobic bottle, BacT/Alert FAN aerobic bottle and BacT/Alert enhanced FAN aerobic bottle in simulated blood culture.

    Science.gov (United States)

    Sümerkan, B; Gökahmetoglu, S; Esel, D

    2001-07-01

    The objective of this study was to compare the performances of the standard aerobic bottle (StAe), FAN aerobic (FANAe) and enhanced FAN aerobic (E-FANAe) (the charcoal component of the FANAe was revised recently to improve the feasibility of Gram smear interpretation) blood culture bottles for BacT/Alert system for the detection of Brucella melitensis in simulated blood culture. Triplicate strains of eight clinical isolates of B. melitensis were studied. Each bottle was inoculated with 5 mL of freshly collected human blood at three different targeted bacterial inocula (10(1), 10(2) and 10(3) CFU/bottle). All bottles were monitored for up to 21 days or until they became positive. The results of time to detection (TTD) on the eight B. melitensis samples were as follows: at 10(1) CFU/bottle, the E-FANAe had a mean TTD significantly shorter than the StAe (48 h vs. 56.2 h, P StAe (41.2 h and 40 h vs. 45.6 h, P StAe, FANAe and E-FANAe were 96, 83 and 58%, respectively. At 10(3) CFU/bottle, the reproducibilities of StAe, FANAe and E-FANAe were 95, 95 and 91%, respectively. Positive results for the presence of bacteria in Gram smears were confirmed in 68% of StAe, 54% of FANAe and 90% of E-FANAe. In case of suspected brucellosis, the combination of one StAe bottle and one E-FANAe bottle seems to provide the highest and fastest recovery of the organism.

  13. Natural Regeneration in a Multi-Layered Pinus sylvestris-Picea abies Forest after Target Diameter Harvest and Soil Scarification

    Directory of Open Access Journals (Sweden)

    Lars Drössler

    2017-01-01

    Full Text Available Forest management in Sweden can be characterized by even-aged silviculture heavily relying on three established harvest regimes: clearcutting, the seed-tree method, and the shelterwood system. Less intense, small-scale retention harvest systems such as single tree and group selection harvest are rarely used. In addition, natural regeneration dynamics without enrichment planting have barely been studied. Consequently, this study examined natural regeneration establishment in a multi-layered Pinus sylvestris-Picea abies forest stand in southwest Sweden after target diameter harvesting and soil scarification. The creation of forest canopy gaps had a positive effect on total seedling density five years after harvest, mainly due to a significantly higher number of Betula pendula individuals. Seedling density of more desirable tree species suitable for continuous cover forestry such as Fagus sylvatica, Quercus petraea and Picea abies also increased substantially in gaps when compared to pre-harvest conditions or the unharvested plots. In contrast, soil scarification did not increase the number of seedlings of desired tree species due to a significant decrease in Picea abies abundance. Soil moisture and gap size significantly improved Betula pendula seedling establishment while a larger number of Quercus petraea seedlings were observed in Vaccinium myrtillus patches. We conclude that canopy gaps are beneficial under the encountered stand conditions to initiate forest regeneration, and that soil scarification without the timely occurrence of a mast year of desired tree species is not effective in the type of forest studied.

  14. Genome-wide BAC-end sequencing of Musa acuminata DH Pahang reveals further insights into the genome organization of banana

    NARCIS (Netherlands)

    Arnago, R.E.; Togawa, R.C.; Carpentier, S.C.; Lintel Hekkert, te B.; Kema, G.H.J.; Souza, M.T.

    2011-01-01

    Banana and plantain (Musa spp.) are grown in more than 120 countries in tropical and subtropical regions and constitute an important staple food for millions of people. A Musa acuminata ssp. malaccencis DH Pahang bacterial artificial chromosome (BAC) library (MAMB) was submitted for BAC-end

  15. Terpene chemodiversity of relict conifers Picea omorika, Pinus heldreichii, and Pinus peuce, endemic to Balkan.

    Science.gov (United States)

    Nikolić, Biljana; Ristić, Mihailo; Tešević, Vele; Marin, Petar D; Bojović, Srdjan

    2011-12-01

    Terpenes are often used as ecological and chemotaxonomic markers of plant species, as well as for estimation of geographic variability. Essential oils of relic and Balkan endemic/subendemic conifers, Picea omorika, Pinus heldreichii, and P. peuce, in central part of Balkan Peninsula (Serbia and Montenegro), on the level of terpene classes and common terpene compounds were investigated. In finding terpene combinations, which could show the best diversity between species and their natural populations, several statistical methods were applied. Apart from the content of different terpene classes (P. omorika has the most abundant O-containing monoterpenes and sesquiterpenes; P. heldreichii and P. peuce have the largest abundance of sesquiterpene and monoterpene hydrocarbons, resp.), the species are clearly separated according to terpene profile with 22 common compounds. But, divergences in their populations were established only in combination of several compounds (specific for each species), and they were found to be the results of geomorphologic, climatic, and genetic factors. We found similarities between investigated species and some taxa from literature with respect to terpene composition, possibly due to hybridization and phylogenetic relations. Obtained results are also important regarding to chemotaxonomy, biogeography, phylogeny, and evolution of these taxa. Copyright © 2011 Verlag Helvetica Chimica Acta AG, Zürich.

  16. Predicting adaptive phenotypes from multilocus genotypes in Sitka spruce (Picea sitchensis) using random forest.

    Science.gov (United States)

    Holliday, Jason A; Wang, Tongli; Aitken, Sally

    2012-09-01

    Climate is the primary driver of the distribution of tree species worldwide, and the potential for adaptive evolution will be an important factor determining the response of forests to anthropogenic climate change. Although association mapping has the potential to improve our understanding of the genomic underpinnings of climatically relevant traits, the utility of adaptive polymorphisms uncovered by such studies would be greatly enhanced by the development of integrated models that account for the phenotypic effects of multiple single-nucleotide polymorphisms (SNPs) and their interactions simultaneously. We previously reported the results of association mapping in the widespread conifer Sitka spruce (Picea sitchensis). In the current study we used the recursive partitioning algorithm 'Random Forest' to identify optimized combinations of SNPs to predict adaptive phenotypes. After adjusting for population structure, we were able to explain 37% and 30% of the phenotypic variation, respectively, in two locally adaptive traits--autumn budset timing and cold hardiness. For each trait, the leading five SNPs captured much of the phenotypic variation. To determine the role of epistasis in shaping these phenotypes, we also used a novel approach to quantify the strength and direction of pairwise interactions between SNPs and found such interactions to be common. Our results demonstrate the power of Random Forest to identify subsets of markers that are most important to climatic adaptation, and suggest that interactions among these loci may be widespread.

  17. Resonance wood [Picea abies (L.) Karst.]--evaluation and prediction of violin makers' quality-grading.

    Science.gov (United States)

    Buksnowitz, Christoph; Teischinger, Alfred; Müller, Ulrich; Pahler, Andreas; Evans, Robert

    2007-04-01

    The definition of quality in the field of resonance wood for musical instrument making has attracted considerable interest over decades but has remained incomplete. The current work compares the traditional knowledge and practical experience of violin makers with a material-science approach to objectively characterize the properties of resonance wood. Norway spruce [Picea abies (L.) Karst.] has earned a very high reputation for the construction of resonance tops of stringed instruments and resonance boards of keyboard instruments, and was therefore chosen as the focus of the investigation. The samples were obtained from numerous renowned resonance wood regions in the European Alps and cover the whole range of available qualities. A set of acoustical, anatomical, mechanical and optical material properties was measured on each sample. These measurements were compared with subjective quality grading by violin makers, who estimated the acoustical, optical and overall suitability for violin making. Multiple linear regression models were applied to evaluate the predictability of the subjective grading using the measured material characteristics as predictors. The results show that luthiers are able to estimate wood quality related to visible features, but predictions of mechanical and acoustical properties proved to be very poor.

  18. Experimental versus modelled water use in mature Norway spruce (Picea abies exposed to elevated CO2

    Directory of Open Access Journals (Sweden)

    Sebastian eLeuzinger

    2012-10-01

    Full Text Available Rising levels of atmospheric CO2 have often been reported to reduce plant water use. Such behaviour is also predicted by standard equations relating photosynthesis, stomatal conductance, and atmospheric CO2 concentration, which form the core of global dynamic vegetation models (DGVMs. Here, we provide first results from a free air CO2 enrichment (FACE experiment with naturally growing, mature (35 m Picea abies (L. (Norway spruce and compare them to simulations by the DGVM LPJ-GUESS. We monitored sap flow, stem water deficit, stomatal conductance, leaf water potential and soil moisture in five 35-40 m tall CO2-treated (550 ppm trees over two seasons. Using LPJ-GUESS, we simulated this experiment using climate data from a nearby weather station. While the model predicted a stable reduction of transpiration of between 9 and 18 % (at concentrations of 550-700ppm atmospheric CO2, the combined evidence from various methods characterising water use in our experimental trees suggest no changes in response to future CO2 concentrations. The discrepancy between the modelled and the experimental results may be a scaling issue: while dynamic vegetation models correctly predict leaf-level responses, they may not sufficiently account for the processes involved at the canopy and ecosystem scale, which could mitigate the first-order stomatal response.

  19. Extraction of antioxidants from spruce (Picea abies) bark using eco-friendly solvents.

    Science.gov (United States)

    Co, Michelle; Fagerlund, Amelie; Engman, Lars; Sunnerheim, Kerstin; Sjöberg, Per J R; Turner, Charlotta

    2012-01-01

    Antioxidants are known to avert oxidation processes and they are found in trees and other plant materials. Tree bark is a major waste product from paper pulp industries; hence it is worthwhile to develop an extraction technique to extract the antioxidants. To develop a fast and environmentally sustainable extraction technique for the extraction of antioxidants from bark of spruce (Picea abies) and also to identify the extracted antioxidants that are abundant in spruce bark. A screening experiment that involved three different techniques was conducted to determine the best technique to extract antioxidants. The antioxidant capacity of the extracts was determined with DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Pressurised fluid extraction (PFE) turned out to be the best technique and a response surface design was therefore utilised to optimise PFE. Furthermore, NMR and HPLC-DAD-MS/MS were applied to identify the extracted antioxidants. PFE using water and ethanol as solvent at 160 and 180°C, respectively, gave extracts of the highest antioxidant capacity. Stilbene glucosides such as isorhapontin, piceid and astringin were identified in the extracts. The study has shown that PFE is a fast and environmentally sustainable technique, using water and ethanol as solvent for the extraction of antioxidants from spruce bark. Copyright © 2011 John Wiley & Sons, Ltd.

  20. Patterns of nucleotide diversity at photoperiod related genes in Norway spruce [Picea abies (L.) Karst].

    Science.gov (United States)

    Källman, Thomas; De Mita, Stéphane; Larsson, Hanna; Gyllenstrand, Niclas; Heuertz, Myriam; Parducci, Laura; Suyama, Yoshihisa; Lagercrantz, Ulf; Lascoux, Martin

    2014-01-01

    The ability of plants to track seasonal changes is largely dependent on genes assigned to the photoperiod pathway, and variation in those genes is thereby important for adaptation to local day length conditions. Extensive physiological data in several temperate conifer species suggest that populations are adapted to local light conditions, but data on the genes underlying this adaptation are more limited. Here we present nucleotide diversity data from 19 genes putatively involved in photoperiodic response in Norway spruce (Picea abies). Based on similarity to model plants the genes were grouped into three categories according to their presumed position in the photoperiod pathway: photoreceptors, circadian clock genes, and downstream targets. An HKA (Hudson, Kreitman and Aquade) test showed a significant excess of diversity at photoreceptor genes, but no departure from neutrality at circadian genes and downstream targets. Departures from neutrality were also tested with Tajima's D and Fay and Wu's H statistics under three demographic scenarios: the standard neutral model, a population expansion model, and a more complex population split model. Only one gene, the circadian clock gene PaPRR3 with a highly positive Tajima's D value, deviates significantly from all tested demographic scenarios. As the PaPRR3 gene harbours multiple non-synonymous variants it appears as an excellent candidate gene for control of photoperiod response in Norway spruce.

  1. Warming delays autumn declines in photosynthetic capacity in a boreal conifer, Norway spruce (Picea abies).

    Science.gov (United States)

    Stinziano, Joseph R; Hüner, Norman P A; Way, Danielle A

    2015-12-01

    Climate change, via warmer springs and autumns, may lengthen the carbon uptake period of boreal tree species, increasing the potential for carbon sequestration in boreal forests, which could help slow climate change. However, if other seasonal cues such as photoperiod dictate when photosynthetic capacity declines, warmer autumn temperatures may have little effect on when carbon uptake capacity decreases in these species. We investigated whether autumn warming would delay photosynthetic decline in Norway spruce (Picea abies (L.) H. Karst.) by growing seedlings under declining weekly photoperiods and weekly temperatures either at ambient temperature or a warming treatment 4 °C above ambient. Photosynthetic capacity was relatively constant in both treatments when weekly temperatures were >8 °C, but declined rapidly at lower temperatures, leading to a delay in the autumn decline in photosynthetic capacity in the warming treatment. The decline in photosynthetic capacity was not related to changes in leaf nitrogen or chlorophyll concentrations, but was correlated with a decrease in the apparent fraction of leaf nitrogen invested in Rubisco, implicating a shift in nitrogen allocation away from the Calvin cycle at low autumn growing temperatures. Our data suggest that as the climate warms, the period of net carbon uptake will be extended in the autumn for boreal forests dominated by Norway spruce, which could increase total carbon uptake in these forests. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  2. Soil physical, chemical and gas-flux characterization from Picea mariana stands near Erickson Creek, Alaska

    Science.gov (United States)

    O'Donnell, Jonathan A.; Harden, Jennifer W.; Manies, Kristen L.

    2011-01-01

    Fire is a particularly important control on the carbon (C) balance of the boreal forest, and fire-return intervals and fire severity appear to have increased since the late 1900s in North America. In addition to the immediate release of stored C to the atmosphere through organic-matter combustion, fire also modifies soil conditions, possibly affecting C exchange between terrestrial and atmospheric pools for decades after the burn. The effects of fire on ecosystem C dynamics vary across the landscape, with topographic position and soil drainage functioning as important controls. The data reported here contributed to a larger U.S. Geological Survey (USGS) study, published in the journal Ecosystems by O'Donnell and others (2009). To evaluate the effects of fire and drainage on ecosystem C dynamics, we selected sample sites within the 2003 Erickson Creek fire scar to measure CO2 fluxes and soil C inventories in burned and unburned (control) sites in both upland and lowland black spruce (Picea mariana) forests. The results of this study suggested that although fire can create soil climate conditions which are more conducive to rapid decomposition, rates of C release from soils may be constrained after fire by changes in moisture and (or) substrate quality that impede rates of decomposition. Here, we report detailed site information, methodology, and data (in spreadsheet files) from that study.

  3. Evaluation of the stomatal conductance formulation in the EMEP ozone deposition model for Picea abies

    Science.gov (United States)

    Wieser, G.; Emberson, L. D.

    It is widely acknowledged that the possible impacts of ozone on forest trees are more closely related to ozone flux through the stomata than to external ozone exposure. However, the application of the flux approach on a European scale requires the availability of appropriate models, such as the European Monitoring and Evaluation Programme (EMEP) ozone deposition model, for estimating ozone flux and cumulative ozone uptake. Within this model stomatal conductance is the key variable, since it determines the amount of ozone absorbed by the leaves. This paper describes the suitability of the existing EMEP ozone deposition model parameterisation and formulation to represent stomatal behaviour determined from field measurements on adult Norway spruce ( Picea abies (L.) Karst.) trees in the Central European Alps. Parameters affecting maximum stomatal conductance (e.g. seasonal phenology, needle position, needle age, nutrient deficiency and ozone itself) and stomatal response functions to temperature, irradiance, vapour pressure deficit, and soil water content are investigated. Finally, current limitations and possible alterations of the EMEP model will be discussed with respect to spatial scales of available input data for future flux modelling.

  4. The dependence of water potential in shoots of Picea abies on air and soil water status

    Directory of Open Access Journals (Sweden)

    A. Sellin

    Full Text Available Where there is sufficient water storage in the soil the water potential (Ψx in shoots of Norway spruce [Picea abies (L. Karst.] is strongly governed by the vapour pressure deficit of the atmosphere, while the mean minimum values of Ψx usually do not drop below –1.5 MPa under meteorological conditions in Estonia. If the base water potential (Ψb is above –0.62 MPa, the principal factor causing water deficiency in shoots of P. abies may be either limited soil water reserves or atmospheric evaporative demand depending on the current level of the vapour pressure deficit. As the soil dries the stomatal control becomes more efficient in preventing water losses from the foliage, and the leaf water status, in turn, less sensitive to atmospheric demand. Under drought conditions, if Ψb falls below –0.62 MPa, the trees' water stress is mainly caused by low soil water availability. Further declines in the shoot water potential (below –1.5 MPa can be attributed primarily to further decreases in the soil water, i.e. to the static water stress.Key words. Hydrology (evapotranspiration · plant ecology · soil moisture.

  5. GENETIC STRUCTURE OF NORWAY SPRUCE (PICEA ABIES): CONCORDANCE OF MORPHOLOGICAL AND ALLOZYMIC VARIATION.

    Science.gov (United States)

    Lagercrantz, Ulf; Ryman, Nils

    1990-02-01

    This study describes the population structure of Norway spruce (Picea abies) as revealed by protein polymorphisms and morphological variation. Electrophoretically detectable genetic variability was examined at 22 protein loci in 70 populations from the natural range of the species in Europe. Like other conifers, Norway spruce exhibits a relatively large amount of genetic variability and little differentiation among populations. Sixteen polymorphic loci (73%) segregate for a total of 51 alleles, and average heterozygosity per population is 0.115. Approximately 5% of the total genetic diversity is explained by differences between populations (G ST = 0.052), and Nei's standard genetic distance is less than 0.04 in all cases. We suggest that the population structure largely reflects relatively recent historical events related to the last glaciation and that Norway spruce is still in a process of adaptation and differentiation. There is a clear geographic pattern in the variation of allele frequencies. A major part of the allelefrequency variation can be accounted for by a few synthetic variables (principal components), and 80% of the variation of the first principal component is "explained" by latitude and longitude. The central European populations are consistently depauperate of genetic variability, most likely as an effect of severe restrictions of population size during the last glaciation. The pattern of differentiation at protein loci is very similar to that observed for seven morphological traits examined. This similarity suggests that the same evolutionary forces have acted upon both sets of characters. © 1990 The Society for the Study of Evolution.

  6. Review of Lepidoptera with trophic relationships to Picea abies (L. in the conditions of Czechia

    Directory of Open Access Journals (Sweden)

    Modlinger Roman

    2016-12-01

    Full Text Available Trophic relationships of Lepidoptera (Insecta occurring in the territory of Czechia to the Norway spruce (Picea abies L. was evaluated on the basis of the excerption and critical evaluation of literature. Each species was classified into the following categories – spruce as the host plant, regular development on spruce, narrow trophic relationship, indirect relationship and episodical occurrence. The particular taxa were also characterized according to their distribution and the form of larval life was specified. The development on spruce was documented in 96 species of Lepidoptera, which represented less than 3% of taxa belonging to this group and being reported from Czechia. Of that, spruce was a common host plant for 67 species, 23 species were polyphagous and might develop on spruce, and 6 species belonged to soil species damaging spruce roots, mainly in forest nurseries. Among the species of Lepidoptera, which regularly develop on spruce in the Czech conditions, 55 species were classified. As narrow specialists with special trophic relationship to spruce, 33 taxa could be considered. There were 15 spruce species with forestry importance, which were able to outbreak their populations regularly or irregularly. Among spruce species it was possible to classify 16 taxa as rare. The provided information on Lepidoptera with trophic relationship to spruce is applicable also for other Central European areas. Besides the species with importance for forest pest management, also rare taxa, which can become endangered by climate change or by forest management, were indicated.

  7. An overview of the Phalaenopsis orchid genome through BAC end sequence analysis

    Directory of Open Access Journals (Sweden)

    Hsiao Yu-Yun

    2011-01-01

    Full Text Available Abstract Background Phalaenopsis orchids are popular floral crops, and development of new cultivars is economically important to floricultural industries worldwide. Analysis of orchid genes could facilitate orchid improvement. Bacterial artificial chromosome (BAC end sequences (BESs can provide the first glimpses into the sequence composition of a novel genome and can yield molecular markers for use in genetic mapping and breeding. Results We used two BAC libraries (constructed using the BamHI and HindIII restriction enzymes of Phalaenopsis equestris to generate pair-end sequences from 2,920 BAC clones (71.4% and 28.6% from the BamHI and HindIII libraries, respectively, at a success rate of 95.7%. A total of 5,535 BESs were generated, representing 4.5 Mb, or about 0.3% of the Phalaenopsis genome. The trimmed sequences ranged from 123 to 1,397 base pairs (bp in size, with an average edited read length of 821 bp. When these BESs were subjected to sequence homology searches, it was found that 641 (11.6% were predicted to represent protein-encoding regions, whereas 1,272 (23.0% contained repetitive DNA. Most of the repetitive DNA sequences were gypsy- and copia-like retrotransposons (41.9% and 12.8%, respectively, whereas only 10.8% were DNA transposons. Further, 950 potential simple sequence repeats (SSRs were discovered. Dinucleotides were the most abundant repeat motifs; AT/TA dimer repeats were the most frequent SSRs, representing 253 (26.6% of all identified SSRs. Microsynteny analysis revealed that more BESs mapped to the whole-genome sequences of poplar than to those of grape or Arabidopsis, and even fewer mapped to the rice genome. This work will facilitate analysis of the Phalaenopsis genome, and will help clarify similarities and differences in genome composition between orchids and other plant species. Conclusion Using BES analysis, we obtained an overview of the Phalaenopsis genome in terms of gene abundance, the presence of repetitive

  8. Construction of an integrated genetic linkage map for the A genome of Brassica napus using SSR markers derived from sequenced BACs in B. rapa

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    King Graham J

    2010-10-01

    Full Text Available Abstract Background The Multinational Brassica rapa Genome Sequencing Project (BrGSP has developed valuable genomic resources, including BAC libraries, BAC-end sequences, genetic and physical maps, and seed BAC sequences for Brassica rapa. An integrated linkage map between the amphidiploid B. napus and diploid B. rapa will facilitate the rapid transfer of these valuable resources from B. rapa to B. napus (Oilseed rape, Canola. Results In this study, we identified over 23,000 simple sequence repeats (SSRs from 536 sequenced BACs. 890 SSR markers (designated as BrGMS were developed and used for the construction of an integrated linkage map for the A genome in B. rapa and B. napus. Two hundred and nineteen BrGMS markers were integrated to an existing B. napus linkage map (BnaNZDH. Among these mapped BrGMS markers, 168 were only distributed on the A genome linkage groups (LGs, 18 distrubuted both on the A and C genome LGs, and 33 only distributed on the C genome LGs. Most of the A genome LGs in B. napus were collinear with the homoeologous LGs in B. rapa, although minor inversions or rearrangements occurred on A2 and A9. The mapping of these BAC-specific SSR markers enabled assignment of 161 sequenced B. rapa BACs, as well as the associated BAC contigs to the A genome LGs of B. napus. Conclusion The genetic mapping of SSR markers derived from sequenced BACs in B. rapa enabled direct links to be established between the B. napus linkage map and a B. rapa physical map, and thus the assignment of B. rapa BACs and the associated BAC contigs to the B. napus linkage map. This integrated genetic linkage map will facilitate exploitation of the B. rapa annotated genomic resources for gene tagging and map-based cloning in B. napus, and for comparative analysis of the A genome within Brassica species.

  9. Ecological patterns, distribution and population structure of Prionace glauca (Chondrichthyes: Carcharhinidae) in the tropical-subtropical transition zone of the north-eastern Pacific.

    Science.gov (United States)

    Vögler, Rodolfo; Beier, Emilio; Ortega-García, Sofía; Santana-Hernández, Heriberto; Valdez-Flores, J Javier

    2012-02-01

    Regional ecological patterns, distribution and population structure of Prionace glauca were analyzed based on samples collected on-board two long-line fleets operating in oceanic waters (1994-96/2000-02) and in coastal oceanic waters (2003-2009) of the eastern tropical Pacific off México. Generalized additive models were applied to catch per unit of effort data to evaluate the effect of spatial, temporal and environmental factors on the horizontal distribution of the life stages (juvenile, adult) and the sexes at the estimated depth of catch. The presence of breeding areas was explored. The population structure was characterized by the presence of juveniles' aggregations and pregnant females towards coastal waters and the presence of adult males' aggregations towards oceanic waters. The species exhibited horizontal segregation by sex-size and vertical segregation by sex. Distribution of the sex-size groups at oceanic waters was seasonally affected by the latitude; however, at coastal oceanic waters mainly females were influenced by the longitude. Latitudinal changes on the horizontal distribution were coupled to the seasonal forward and backward of water masses through the study area. Adult males showed positive relationship with high temperatures and high-salinities waters (17.0°-20.0 °C; 34.2-34.4) although they were also detected in low-salinities waters. The distribution of juvenile males mainly occurred beyond low temperatures and low-salinities waters (14.0°-15.0 °C; 33.6-34.1), suggesting a wide tolerance of adult males to explore subartic and subtropical waters. At oceanic areas, adult females were aggregated towards latitudes ecological key region to the reproductive cycle of P. glauca. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Sequencing of 15622 gene-bearing BACs clarifies the gene-dense regions of the barley genome

    Czech Academy of Sciences Publication Activity Database

    Munoz-Amatriain, M.; Lonardi, S.; Luo, M.C.; Madishetty, K.; Svensson, J.T.; Moscou, M. J.; Wanamaker, S.; Kudrna, D.; Zheng, J.; Šimková, Hana; Doležel, Jaroslav; Grimwood, J.; Mammadov, J.; Close, T.J.

    2015-01-01

    Roč. 84, č. 1 (2015), s. 216-227 ISSN 0960-7412 R&D Projects: GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : Barley * Hordeum vulgare L * BAC sequencing Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.468, year: 2015

  11. Characterization of Extracellular Penicilin G Acylase Produced by A New Local Strain of Bacillus subtilis BAC4

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    SUPARTONO

    2008-06-01

    Full Text Available Penicillin G acylase (PGA which catalyses penicillin G hydrolysis reaction is a key enzyme for the industrial production of penicilin G derivatives used in therapeutics. A new local strain of Bacillus subtilis BAC4 was found capable of producing extracellular PGA. However, characteristics of this extracellular PGA are not known. The goal of this research was to characterize the extracellular PGA produced by B. subtilis BAC4. Enzyme production was carried out by batch fermentation, followed by enzyme purification and characterization of the PGA. The PGA activity was determined by the Kornfeld method, with optimal activity for hydrolysing penicillin G observed at 43 °C and pH 8.5. The activation energy of penicillin G hydrolysis by the PGA of B. subtilis BAC4 was determined as 4.9 kcal.mol−1 and Vmax and Km values were found to be 0.7 μmole.min−1.mg−1 and 3.5 mM respectively. PGA catalytic activity was competitively inhibited by phenylacetic acid with an inhibition constant, Ki(PAA, of 347.2 mM. It was concluded that the extracellular PGA of B. subtilis BAC4 can hydrolyse penicillin G efficiently.

  12. Characterization of Extracellular Penicilin G Acylase Produced by A New Local Strain of Bacillus subtilis BAC4

    Directory of Open Access Journals (Sweden)

    SUPARTONO

    2008-06-01

    Full Text Available Penicillin G acylase (PGA which catalyses penicillin G hydrolysis reaction is a key enzyme for the industrial production of penicilin G derivatives used in therapeutics. A new local strain of Bacillus subtilis BAC4 was found capable of producing extracellular PGA. However, characteristics of this extracellular PGA are not known. The goal of this research was to characterize the extracellular PGA produced by B. subtilis BAC4. Enzyme production was carried out by batch fermentation, followed by enzyme purification and characterization of the PGA. The PGA activity was determined by the Kornfeld method, with optimal activity for hydrolysing penicillin G observed at 43 oC and pH 8.5. The activation energy of penicillin G hydrolysis by the PGA of B. subtilis BAC4 was determined as 4.9 kcal.mol-1 and Vmax and Km values were found to be 0.7 µmole.min-1.mg-1 and 3.5 mM respectively. PGA catalytic activity was competitively inhibited by phenylacetic acid with an inhibition constant, Ki(PAA, of 347.2 mM. It was concluded that the extracellular PGA of B. subtilis BAC4 can hydrolyse penicillin G efficiently.

  13. Creation of a BAC resource to study the structure and evolution of the banana (Musa balbisiana) genome

    Czech Academy of Sciences Publication Activity Database

    Šafář, Jan; Noa-Carrazana, J. C.; Vrána, Jan; Bartoš, Jan; Alkhimova, Olena; Lheureux, F.; Šimková, Hana; Caruana, M. L.; Doležel, Jaroslav; Piffanelli, P.

    2004-01-01

    Roč. 47, - (2004), 1182ů1191 E-ISSN 1480-3321 R&D Projects: GA AV ČR IAA6038201 Grant - others:research contract IAEA(FR) 12230/RBF Institutional research plan: CEZ:AV0Z5038910 Keywords : bacterial artificial chromosome library * banana * BAC-FISH Subject RIV: EB - Genetics ; Molecular Biology

  14. Mapping of Micro-Tom BAC-End Sequences to the Reference Tomato Genome Reveals Possible Genome Rearrangements and Polymorphisms

    Science.gov (United States)

    Asamizu, Erika; Shirasawa, Kenta; Hirakawa, Hideki; Sato, Shusei; Tabata, Satoshi; Yano, Kentaro; Ariizumi, Tohru; Shibata, Daisuke; Ezura, Hiroshi

    2012-01-01

    A total of 93,682 BAC-end sequences (BESs) were generated from a dwarf model tomato, cv. Micro-Tom. After removing repetitive sequences, the BESs were similarity searched against the reference tomato genome of a standard cultivar, “Heinz 1706.” By referring to the “Heinz 1706” physical map and by eliminating redundant or nonsignificant hits, 28,804 “unique pair ends” and 8,263 “unique ends” were selected to construct hypothetical BAC contigs. The total physical length of the BAC contigs was 495, 833, 423 bp, covering 65.3% of the entire genome. The average coverage of euchromatin and heterochromatin was 58.9% and 67.3%, respectively. From this analysis, two possible genome rearrangements were identified: one in chromosome 2 (inversion) and the other in chromosome 3 (inversion and translocation). Polymorphisms (SNPs and Indels) between the two cultivars were identified from the BLAST alignments. As a result, 171,792 polymorphisms were mapped on 12 chromosomes. Among these, 30,930 polymorphisms were found in euchromatin (1 per 3,565 bp) and 140,862 were found in heterochromatin (1 per 2,737 bp). The average polymorphism density in the genome was 1 polymorphism per 2,886 bp. To facilitate the use of these data in Micro-Tom research, the BAC contig and polymorphism information are available in the TOMATOMICS database. PMID:23227037

  15. Cross-species BAC-FISH painting of the tomato and potato chromosome 6 reveals undescribed chromosomal rearrangements

    NARCIS (Netherlands)

    Tang, X.; Szinay, D.; Ramanna, M.S.; Vossen, van der E.A.G.; Datema, E.; Klein Lankhorst, R.M.; Boer, de J.M.; Peters, S.A.; Bachem, C.W.B.; Stiekema, W.J.; Visser, R.G.F.; Jong, de J.H.; Bai, Y.

    2008-01-01

    Ongoing genomics projects of tomato (Solanum lycopersicum ) and potato (Solanum tuberosum) are providing unique tools for comparative mapping studies in Solanaceae. At the chromosomal level, BACs can be positioned on pachytene comple-ments by fluorescent in situ hybridization (FISH) on homoeologous

  16. Development of Health Education Learning Module in Bac.TSE-LDPE Programme in TTI: Needs Analysis Study

    Science.gov (United States)

    Ujang, Alijah; Alias, Norlidah; Siraj, Saedah

    2015-01-01

    This study is to explore the need to develop learning modules of health education for trainee teachers in the Bachelor Of Teaching (Hons)(Special Education-Learning Disabilities For Primary Education) Programme (Bac.TSE-LDPE) in the Teacher Training Institute (TTI). The questionnaire uses the Likert scale with the close ended questions analysed by…

  17. BAC end sequencing of Pacific white shrimp Litopenaeus vannamei: a glimpse into the genome of Penaeid shrimp

    Science.gov (United States)

    Zhao, Cui; Zhang, Xiaojun; Liu, Chengzhang; Huan, Pin; Li, Fuhua; Xiang, Jianhai; Huang, Chao

    2012-05-01

    Little is known about the genome of Pacific white shrimp ( Litopenaeus vannamei). To address this, we conducted BAC (bacterial artificial chromosome) end sequencing of L. vannamei. We selected and sequenced 7 812 BAC clones from the BAC library LvHE from the two ends of the inserts by Sanger sequencing. After trimming and quality filtering, 11 279 BAC end sequences (BESs) including 4 609 pairedends BESs were obtained. The total length of the BESs was 4 340 753 bp, representing 0.18% of the L. vannamei haploid genome. The lengths of the BESs ranged from 100 bp to 660 bp with an average length of 385 bp. Analysis of the BESs indicated that the L. vannamei genome is AT-rich and that the primary repeats patterns were simple sequence repeats (SSRs) and low complexity sequences. Dinucleotide and hexanucleotide repeats were the most common SSR types in the BESs. The most abundant transposable element was gypsy, which may contribute to the generation of the large genome size of L. vannamei. We successfully annotated 4 519 BESs by BLAST searching, including genes involved in immunity and sex determination. Our results provide an important resource for functional gene studies, map construction and integration, and complete genome assembly for this species.

  18. Development of a Set of Chromosome-Specific Cytogenetic DNA Markers in Sunflower Using BAC-FISH

    Science.gov (United States)

    In diploid sunflower (2n=34), conventional karyotypes and various genetic linkage maps have been established. However, the relationship between genetic linkage groups and individual chromosomes of sunflower remains unknown. Recently, a set of linkage group-specific BAC and BIBAC clones were identifi...

  19. A comparative, BAC end sequence enabled map of the genome of the American mink (Neovison vison)

    DEFF Research Database (Denmark)

    Benkel, Bernhard F.; Smith, Amanda; Christensen, Knud

    2012-01-01

    In this report we present the results of the analysis of approximately 2.7 Mb of genomic information for the American mink (Neovison vison) derived through BAC end sequencing. Our study, which encompasses approximately 1/1000th of the mink genome, suggests that simple sequence repeats (SSRs...

  20. Co-oxidation of carcinogenic polycyclic aromatic hydrocarbons with some biologically active compounds (BAC)

    Energy Technology Data Exchange (ETDEWEB)

    Gubergrits, M.Y.

    1978-09-01

    Oxidation of benzo(a)pyrene (BP) initiated by UV or gamma irradiation was promoted by benz(a)anthracene and 7,12-dimethylbenz(a)anthracene (DMBA) and inhibited by pyrene, dibenz(a,c)anthracene, and asymmetric benz(a)antharacene. The effects of these BAC commonly occurring together with BP in industrial wastes, increased with their concentrations. Phenol and 3-methylcholanthrene strongly promoted BP oxidation when present at low concentrations and inhibited it at high concentrations. Consistent promoting effect was also observed in BP co-oxidation with adipic acid, ..cap alpha..-naphthoflavon, and vitamin E, whereas succinic, azelaic, ferulic, gallic, and chlorogenic acids, rutin, and vitamin C acted as inhibitors. Most saturated dicarboxylic acids studied did not affect BP oxidation at 1:1 acid-BP molar ratio. The kinetics of 7,12-DMBA photooxidation inhibition by some metabolic intermediates, e.g., DMBA endo-peroxide, were also studied.

  1. Multicenter Clinical Evaluation of BacT/Alert Virtuo Blood Culture System.

    Science.gov (United States)

    Jacobs, Michael R; Mazzulli, Tony; Hazen, Kevin C; Good, Caryn E; Abdelhamed, Ayman M; Lo, Pauline; Shum, Bianche; Roman, Katharine P; Robinson, Danielle C

    2017-08-01

    BacT/Alert Virtuo is an advanced, automated blood culture system incorporating improved automation and an enhanced detection algorithm to shorten time to detection. A multicenter study of the investigational Virtuo system (bioMérieux, Inc., Durham, NC) compared to BacT/Alert 3D (BTA3D) for detection of bacteremia/fungemia in four bottle types, SA and FA Plus (aerobic) and SN and FN Plus (anaerobic), was performed in a clinical setting with patient samples in a matched system design clinical trial. Blood was added to paired aerobic or anaerobic bottles, with the volume in each bottle in each pair required to be ≤10 ml and with the volumes required to be within 30% of each other. Of 5,709 bottle sets (52.5% aerobic pairs and 47.5% anaerobic pairs), 430 (7.5%) were positive for bacterial or fungal growth, with 342 (6.0%) clinically significant and 83 (1.5%) contaminated. A total of 3,539 sets (62.0%) were volume compliant, with 203 sets (5.7%) clinically significant. The positivity rates for volume-compliant bottle pairs determined by the two systems were comparable, with 68.7% of clinically significant isolates detected by both instruments, 15.7% by Virtuo only, and 15.7% by BTA3D only. Virtuo detected microbial growth nearly 2 h sooner overall than BTA3D (mean, 15.9 h versus 17.7 h). Shorter time to detection by Virtuo was related to organism group, with the time to detection being significantly shorter for enteric Gram-negative bacilli and enterococci (means, 3.6 h and 2.3 h shorter, respectively). This large clinical study demonstrated that the Virtuo blood culture system produced results comparable to those seen with the long-established BTA3D system, with significantly shorter time to detection. Copyright © 2017 Jacobs et al.

  2. A BAC-bacterial recombination method to generate physically linked multiple gene reporter DNA constructs

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    Gong Shiaochin

    2009-03-01

    Full Text Available Abstract Background Reporter gene mice are valuable animal models for biological research providing a gene expression readout that can contribute to cellular characterization within the context of a developmental process. With the advancement of bacterial recombination techniques to engineer reporter gene constructs from BAC genomic clones and the generation of optically distinguishable fluorescent protein reporter genes, there is an unprecedented capability to engineer more informative transgenic reporter mouse models relative to what has been traditionally available. Results We demonstrate here our first effort on the development of a three stage bacterial recombination strategy to physically link multiple genes together with their respective fluorescent protein (FP reporters in one DNA fragment. This strategy uses bacterial recombination techniques to: (1 subclone genes of interest into BAC linking vectors, (2 insert desired reporter genes into respective genes and (3 link different gene-reporters together. As proof of concept, we have generated a single DNA fragment containing the genes Trap, Dmp1, and Ibsp driving the expression of ECFP, mCherry, and Topaz FP reporter genes, respectively. Using this DNA construct, we have successfully generated transgenic reporter mice that retain two to three gene readouts. Conclusion The three stage methodology to link multiple genes with their respective fluorescent protein reporter works with reasonable efficiency. Moreover, gene linkage allows for their common chromosomal integration into a single locus. However, the testing of this multi-reporter DNA construct by transgenesis does suggest that the linkage of two different genes together, despite their large size, can still create a positional effect. We believe that gene choice, genomic DNA fragment size and the presence of endogenous insulator elements are critical variables.

  3. A BAC-bacterial recombination method to generate physically linked multiple gene reporter DNA constructs.

    Science.gov (United States)

    Maye, Peter; Stover, Mary Louise; Liu, Yaling; Rowe, David W; Gong, Shiaochin; Lichtler, Alexander C

    2009-03-13

    Reporter gene mice are valuable animal models for biological research providing a gene expression readout that can contribute to cellular characterization within the context of a developmental process. With the advancement of bacterial recombination techniques to engineer reporter gene constructs from BAC genomic clones and the generation of optically distinguishable fluorescent protein reporter genes, there is an unprecedented capability to engineer more informative transgenic reporter mouse models relative to what has been traditionally available. We demonstrate here our first effort on the development of a three stage bacterial recombination strategy to physically link multiple genes together with their respective fluorescent protein (FP) reporters in one DNA fragment. This strategy uses bacterial recombination techniques to: (1) subclone genes of interest into BAC linking vectors, (2) insert desired reporter genes into respective genes and (3) link different gene-reporters together. As proof of concept, we have generated a single DNA fragment containing the genes Trap, Dmp1, and Ibsp driving the expression of ECFP, mCherry, and Topaz FP reporter genes, respectively. Using this DNA construct, we have successfully generated transgenic reporter mice that retain two to three gene readouts. The three stage methodology to link multiple genes with their respective fluorescent protein reporter works with reasonable efficiency. Moreover, gene linkage allows for their common chromosomal integration into a single locus. However, the testing of this multi-reporter DNA construct by transgenesis does suggest that the linkage of two different genes together, despite their large size, can still create a positional effect. We believe that gene choice, genomic DNA fragment size and the presence of endogenous insulator elements are critical variables.

  4. Genomic insight into the common carp (Cyprinus carpio genome by sequencing analysis of BAC-end sequences

    Directory of Open Access Journals (Sweden)

    Wang Jintu

    2011-04-01

    Full Text Available Abstract Background Common carp is one of the most important aquaculture teleost fish in the world. Common carp and other closely related Cyprinidae species provide over 30% aquaculture production in the world. However, common carp genomic resources are still relatively underdeveloped. BAC end sequences (BES are important resources for genome research on BAC-anchored genetic marker development, linkage map and physical map integration, and whole genome sequence assembling and scaffolding. Result To develop such valuable resources in common carp (Cyprinus carpio, a total of 40,224 BAC clones were sequenced on both ends, generating 65,720 clean BES with an average read length of 647 bp after sequence processing, representing 42,522,168 bp or 2.5% of common carp genome. The first survey of common carp genome was conducted with various bioinformatics tools. The common carp genome contains over 17.3% of repetitive elements with GC content of 36.8% and 518 transposon ORFs. To identify and develop BAC-anchored microsatellite markers, a total of 13,581 microsatellites were detected from 10,355 BES. The coding region of 7,127 genes were recognized from 9,443 BES on 7,453 BACs, with 1,990 BACs have genes on both ends. To evaluate the similarity to the genome of closely related zebrafish, BES of common carp were aligned against zebrafish genome. A total of 39,335 BES of common carp have conserved homologs on zebrafish genome which demonstrated the high similarity between zebrafish and common carp genomes, indicating the feasibility of comparative mapping between zebrafish and common carp once we have physical map of common carp. Conclusion BAC end sequences are great resources for the first genome wide survey of common carp. The repetitive DNA was estimated to be approximate 28% of common carp genome, indicating the higher complexity of the genome. Comparative analysis had mapped around 40,000 BES to zebrafish genome and established over 3

  5. Genomic insight into the common carp (Cyprinus carpio) genome by sequencing analysis of BAC-end sequences

    Science.gov (United States)

    2011-01-01

    Background Common carp is one of the most important aquaculture teleost fish in the world. Common carp and other closely related Cyprinidae species provide over 30% aquaculture production in the world. However, common carp genomic resources are still relatively underdeveloped. BAC end sequences (BES) are important resources for genome research on BAC-anchored genetic marker development, linkage map and physical map integration, and whole genome sequence assembling and scaffolding. Result To develop such valuable resources in common carp (Cyprinus carpio), a total of 40,224 BAC clones were sequenced on both ends, generating 65,720 clean BES with an average read length of 647 bp after sequence processing, representing 42,522,168 bp or 2.5% of common carp genome. The first survey of common carp genome was conducted with various bioinformatics tools. The common carp genome contains over 17.3% of repetitive elements with GC content of 36.8% and 518 transposon ORFs. To identify and develop BAC-anchored microsatellite markers, a total of 13,581 microsatellites were detected from 10,355 BES. The coding region of 7,127 genes were recognized from 9,443 BES on 7,453 BACs, with 1,990 BACs have genes on both ends. To evaluate the similarity to the genome of closely related zebrafish, BES of common carp were aligned against zebrafish genome. A total of 39,335 BES of common carp have conserved homologs on zebrafish genome which demonstrated the high similarity between zebrafish and common carp genomes, indicating the feasibility of comparative mapping between zebrafish and common carp once we have physical map of common carp. Conclusion BAC end sequences are great resources for the first genome wide survey of common carp. The repetitive DNA was estimated to be approximate 28% of common carp genome, indicating the higher complexity of the genome. Comparative analysis had mapped around 40,000 BES to zebrafish genome and established over 3,100 microsyntenies, covering over 50% of

  6. Single-molecule study of full-length NaChBac by planar lipid bilayer recording.

    Directory of Open Access Journals (Sweden)

    Andrew Jo

    Full Text Available Planar lipid bilayer device, alternatively known as BLM, is a powerful tool to study functional properties of conducting membrane proteins such as ion channels and porins. In this work, we used BLM to study the prokaryotic voltage-gated sodium channel (Nav NaChBac in a well-defined membrane environment. Navs are an essential component for the generation and propagation of electric signals in excitable cells. The successes in the biochemical, biophysical and crystallographic studies on prokaryotic Navs in recent years has greatly promoted the understanding of the molecular mechanism that underlies these proteins and their eukaryotic counterparts. In this work, we investigated the single-molecule conductance and ionic selectivity behavior of NaChBac. Purified NaChBac protein was first reconstituted into lipid vesicles, which is subsequently incorporated into planar lipid bilayer by fusion. At single-molecule level, we were able to observe three distinct long-lived conductance sub-states of NaChBac. Change in the membrane potential switches on the channel mainly by increasing its opening probability. In addition, we found that individual NaChBac has similar permeability for Na+, K+, and Ca2+. The single-molecule behavior of the full-length protein is essentially highly stochastic. Our results show that planar lipid bilayer device can be used to study purified ion channels at single-molecule level in an artificial environment, and such studies can reveal new protein properties that are otherwise not observable in in vivo ensemble studies.

  7. Coarse root topology of Norway spruce (Picea abies) and its effects on slope stability

    Science.gov (United States)

    Lith, Aniek; Schmaltz, Elmar; Bogaard, Thom; Keesstra, Saskia

    2017-04-01

    The structural distribution of coarse roots and its beneficial effects on soil reinforcement has widely been assessed. However, it is still not fully understood how topological features of coarse roots (e.g. branching patterns) are affected by slope inclination and further influence the ability of young trees to reinforce soil. This study aims to analyse empirically the impact of slope gradient on the topological development of coarse roots and thus to assess its effects on soil reinforcement. We performed root system excavations on two young Picea abies: tree A on a gently inclined plane (β ≈ 12°) where slope failures are not expected; tree B on a slope (β ≈ 35°) with failure potential. The diameter (d) of the segments between distinct root nodes (root ends, branching locations, direction changes and attachments to stem) of coarse roots (d > 2mm) were measured in situ. The spatial coordinates (x,y,z) of the nodes and surface were measured on a plane raster grid, from which segment length (ls), direction and inclination towards the surface (βr) were derived. Roots and segments were classified into laterals (βr classifications (FSC), to obtain quantitative relations between the topological order and number of segments, total and average ls. The maximal root cohesion (cr) of each segment was assessed using material specific tensile forces (Tr), root area ratio (RAR) and βr, assuming that a potential slip surface would cross the root system parallel to the slope. Laterals depicted the majority of roots (57 %) for tree A orientated rather in upslope direction (76.8 %), whereas tree B showed mostly obliques (54 %) orientated rather in downslope direction (55.4 %). Vertical roots were scarcely observable for both trees. DSC showed a high r2 (> 0.84) for the segments and ls. FSC showed high r2 (> 0.95) for the number of segments and the total length. RAR values of tree B are distributed rather upslope (76.8 % of RARtot), compared to 44.5 % of RARtot for tree A

  8. The role of Nicotiana gluca Graham (paraguayan herbs) as an adjuvant in immunomodulation of Newcastle disease vaccine for broilers Estudo da ação de Nicotiana glauca Graham (erva paraguaia) como coadjuvante em vacina contra a doença de Newcastle em frangos de corte

    OpenAIRE

    Fabiane Pereira Gentilini; Telmo Vidor; Rogério Freitag; Marcos Antônio Anciuti; Caren Gularte Quincozes; Marlete Brum Cleff; Geferson Fischer; Carlos Eduardo Nogueira

    2008-01-01

    The Nicotiana glauca is a native plant species from Argentina, but found all over South América, being used against headaches, rheumatism, injuries, ulcers, and so on. Researchers have considered it as having immunomodulation effect. This study was conducted to investigate the use of a aqueous extract of Nicotiana glauca Graham as an immunomodulator (adjuvant) of a Newcastle disease vaccine.. A total of 56 broilers were distributed into 4 ...

  9. Physiological effects of autotoxicity due to DHAP stress on Picea schrenkiana regeneration.

    Directory of Open Access Journals (Sweden)

    Li Yang

    Full Text Available Picea Schrenkiana as one of the most important zonal vegetation was an endemic species in Middle Asia. Natural regeneration of P. Schrenkiana is a long existing problem troubling scientists. The autotoxicity of 3,4-dihydroxy-acetophenone (DHAP was found to be a causative factor causing the failure of P. Schrenkiana natural regeneration. The effects of concentrations of DHAP treatment on the viability of root cell, activities of antioxidant enzymes and levels of P. Schrenkiana phytohormones were performed to disclose the physiological mechanism of DHAP autotoxicity. It was observed that high concentration of DHAP could inhibit the seed germination and seedling growth, but had a hormesis at low concentrations. Analyses showed that the root cells significantly lost their viability treated with high DHAP. The enzymes activities of seedlings were significantly stimulated by the treatment of 0.5 mM DHAP to give a transient increase and then decrease as DHAP concentration increased to 1.0 mM except for GR (glutathione reductase in which DHAP treatment had little effect on its activity. Comparing with the control, an increase in the levels of phytohormones ZT (zeatin, GA3 (gibberellic acid and IAA (indole acetic acid was induced by the treatment of DHAP at low concentrations (0.1-0.25 mM, but the significant deficiency was found treated by high concentrations (0.5-1.0 mM. In addition, the ABA (abscisic acid level increased in all experimental observations. These results suggested that DHAP significantly affected indices of growth and physiology, and provided some new information about different effect in P. Schrenkiana treated with DHAP.

  10. Heterobasidion annosum root rot in Picea abies: Variability in aggressiveness and resistance

    Energy Technology Data Exchange (ETDEWEB)

    Swedjemark, G.

    1995-12-31

    In greenhouse experiments 3-4 year-old plants of Picea abies and Pinus sylvestris were inoculated with the root rot fungus Heterobasidion annosum. The growth rates of 25 S and 14 P isolates of the fungus were measured in the sapwood of about 25 rametes each of 98 Norway spruce clones and in 150 seedlings each of Norway spruce and Scots pine. Correlations were determined between fungal sapwood growth and a variety of variables, i.e. the physical stage, growth rhythm, size and condition of the cuttings, provenance and the growth capacity of the clones estimated in field tests. Genetic parameters such as phenotypic and genotypic variance, genotypic coefficient of variance and broad sense heritability were calculated. H. annosum infection incidence was close to 100 % for inoculated Norway spruce clones, and the mortality rate was about 1 %. Fungal growth in sapwood differed significantly among clones and among fungal isolates. The genotypic correlation coefficient was large (30 %) and broad sense heritability was 0.35. This suggests that good selection progress can be achieved in breeding programs. The physical stage (bud-flush, vegetative and post bud-set stage) of the clones contributed significantly to the total variation in fungal growth. The population structure of H. annosum was studied in two 25-year-old, first generation stands of Norway spruce. The stands had been thinned one and seven years earlier, respectively. All stumps and remaining trees were sampled for H. annosum isolates. The isolates were tested with somatic incompatibility to detect single genets and the distribution of genets in the stand was determined. About 70 % of the 7-year-old thinning stumps were colonized by H, annosum, and 50 % of these stumps were colonized by more than one fungal genet. Sixty-three percent of the remaining trees were infected with H. annosum but among them there was only one genet per tree. 104 refs, 12 figs

  11. CLIMATE CHANGE AND ORIENTAL SPRUCE (PICEA ORIENTALIS ECOSYSTEMS IN EASTERN BLACKSEA REGION OF TURKEY

    Directory of Open Access Journals (Sweden)

    Aydın Tüfekçioğlu

    2008-04-01

    Full Text Available Climate change has been getting more attention from scientific community recently. Eastern Black Sea Region of Turkey will get significant influences from the climate change according to regional climate model (RegCM3. Oriental spruce (Picea orientalis L. is an important tree species of Turkey and it only grows in the Eastern Black Sea Region of Turkey. With the increase in global warming, spruce forests started to have serious bark beetle problems. More than 200 000 trees died in the region recently due to bark beetle attack. We used existing literature related to oriental spruce and future climate of the region and field observations done in the different times to assess current status of the spruce stands. Future climate of the region has been predicted using RegCM3 regional climate model. Climate change could significantly influence distribution, diversity, structure and stability of the oriental spruce ecosystems. According to RegCM3 regional climate model, the temperatures will increase 2-4 °C in the region in the next century. Future climate scenarios predict 200-300 mm increases in precipitation in the eastern part of the region while the western part won't have any increase in precipitation in the next century. Temperature increases in the western part of the region can cause more stress on spruce trees and would probably increase bark beetle attacks. Also, fire could become an important threat in the western part of the region. It is possible to observe 400-800 m upward shift in the spruce belt in the western part. Treeline of spruce stands would probably move upward both in western and eastern part of the North-eastern Blacksea Region.

  12. Does carbon availability control temporal dynamics of radial growth in Norway spruce (Picea abies)?

    Science.gov (United States)

    Oberhuber, Walter; Gruber, Andreas; Swidrak, Irene

    2015-04-01

    Intra-annual dynamics of cambial activity and wood formation of coniferous species exposed to soil dryness revealed early culmination of maximum growth in late spring prior to occurrence of more favourable environmental conditions, i.e., repeated high rainfall events during summer (Oberhuber et al. 2014). Because it is well known that plants can adjust carbon allocation patterns to optimize resource uptake under prevailing environmental constraints, we hypothesize that early decrease in radial stem growth is an adaptation to cope with drought stress, which might require an early switch of carbon allocation to belowground organs. Physical blockage of carbon transport in the phloem through girdling causes accumulation and depletion of carbohydrates above and below the girdle, respectively, making this method quite appropriate to investigate carbon relationships in trees. Hence, in a common garden experiment we will manipulate the carbon status of Norway spruce (Picea abies) saplings by phloem blockage at different phenological stages during the growing season. We will present the methodological approach and first results of the study aiming to test the hypothesis that carbon status of the tree affects temporal dynamics of cambial activity and wood formation in conifers under drought. Acknowledgment The research is funded by the Austrian Science Fund (FWF): P25643-B16 "Carbon allocation and growth of Scots pine". Reference Oberhuber W, A Gruber, W Kofler, I Swidrak (2014) Radial stem growth in response to microclimate and soil moisture in a drought-prone mixed coniferous forest at an inner Alpine site. Eur J For Res 133:467-479.

  13. INVESTIGATION OF THE ESSENTIAL OIL OF EUROPEAN SPRUCE ROOTS (LAT. PICEA ABIES H. KARST, PINACEAE FAMILY

    Directory of Open Access Journals (Sweden)

    D. K. Guljaev

    2017-01-01

    Full Text Available The article is devoted to the study of essential oil extracted from the roots of European Spruce (Lat. Picea abies.The aim is to establish the component composition of the essential oil and the peculiarities of its localization in the roots of European Spruce.Materials and methods. The objects of the study are the roots of European Spruce not longer than two centimeters in diameter, peeled and dried. The study of their anatomical signs was carried out according to the methodology of the State Pharmacopoeia of the Russian Federation (the XIII-th edition with the”Biomed-6” microscope using the DCN 510 nozzle. The essential oil was obtained by hydrodistillation of European Spruce roots using the device of Clevenger by Method 2 of the State Pharmacopoeia of the Russian Federation (the XIII-th edition. The component composition of the essential oil was determined by an Agilent 7890A gas-liquid chromatograph with an Agilent 5975C mass-selective detector.Results and discussion. As a result of the microscopic examination of the roots of European Spruce, it was established that the essential oil is localized mainly in resinous courses located in the wood of the root. In the central part of the root, resin moves are of a larger diameter. More than 18 components were found in the essential oil of European Spruce roots, 14 of them were identifi ed. The main component of the essential oil is sesquiterpene lactone – tanbergol.Conclusion. The essential oil of European Spruce roots has a unique component composition that includes components not characteristic for the essential oil of spruce greenery. The difference in the component composition indicates the difference in properties and pharmacological activity. Further studies are of interest for determining the prospects of using European Spruce roots.

  14. Multi-Objective Optimal Design of Renewable Energy Integrated CCHP System Using PICEA-g

    Directory of Open Access Journals (Sweden)

    Guozheng Li

    2018-03-01

    Full Text Available The integration of renewable energies into combined cooling, heating, and power (CCHP systems has become increasingly popular in recent years. However, the optimization of renewable energies integrated CCHP (RECCHP systems (i.e., optimal component configurations is far from being well addressed, especially in isolated mode. This study aims to fill this research gap. A multi-objective optimization model characterizing the system reliability, system cost, and environmental sustainability is constructed. In this model, the objectives include minimization of annual total cost (ATC, carbon dioxide emission (CDE, and loss of energy supply probability (LESP. The decision variables representing the configuration of the RECCHP system include the number of photovoltaic (PV panels and wind turbines (WTs, the tilt angle of PV panels, the height of WTs, the maximum fuel consumption, and the capacity of battery and heat storage tanks (HSTs. The multi-objective model is solved by a multi-objective evolutionary algorithm, namely, the preference-inspired coevolutionary algorithm (PICEA-g, resulting in a set of Pareto optimal (trade-off solutions. Then, a decision-making process is demonstrated, selecting a preferred solution amongst those trade-off solutions by further considering the decision-maker preferences. Furthermore, on the optimization of the RECCHP system, operational strategies (i.e., following electric load, FEL, and following thermal load, FTL are considered, respectively. Experimental results show that the FEL and FTL strategies lead to different optimal configurations. In general, the FTL is recommended in summer and winter, while the FEL is more suitable for spring and autumn. Compared with traditional energy systems, RECCHP has better economic and environmental advantages.

  15. Mercúrio total em músculo de cação Prionace glauca (Linnaeus, 1758 e de espadarte Xiphias gladius Linnaeus, 1758, na costa sul-sudeste do Brasil e suas implicações para a saúde pública Total mercury in muscle of the shark Prionace glauca (Linnaeus, 1758 and swordfish Xiphias gladius Linnaeus, 1758, from the South-Southeast coast of Brazil and the implications for public health

    Directory of Open Access Journals (Sweden)

    Adriana C. L. Dias

    2008-09-01

    Full Text Available Foram analisadas as concentrações de mercúrio total (THg em tecido muscular do tubarão azul Prionace glauca e do teleósteo Xiphias gladius, vulgarmente conhecido como espadarte, provenientes das regiões sul e sudeste da costa brasileira, para verificar se estas se encontram dentro dos padrões legais para consumo humano. As amostras foram obtidas utilizando-se o programa REVIZEE, de agosto a setembro de 2001, e por intermédio de uma empresa de pesca em Itajaí, Santa Catarina. Foi analisado um total de 95 espécimes, testando-se as correlações entre THg, comprimento (cm e peso (kg. As concentrações de mercúrio total em todas as amostras variaram de 0,13 a 2,26µgg-1 (peso úmido. A média de mercúrio total em P. glauca foi de 0,76 ± 0,48µgg-1 (p.u., e em X. gladius foi de 0,62 ± 0,31µgg-1 (p.u. com diferença não significativa (teste Mann-Whitney, p Total mercury (THg was analyzed in muscle tissue from the blue shark Prionace glauca and the swordfish Xiphias gladius, obtained from the South and Southeast coast of Brazil, to verify compliance with current limits for human consumption. Samples were obtained through the REVIZEE Program and a commercial fishery in Itajaí, Santa Catarina State. A total of 95 specimens were analyzed (48 X. gladius and 47 P. glauca, and correlations were checked between THg and fish length and weight. THg ranged from 0.13 to 2.26µgg-1 (fresh weight, and there was no significant difference between the means for P. glauca, 0.76 ± 0.48µgg-1 (f.w. and X. gladius, 0.62 ± 0.31 (Mann-Whitney test, p < 0.05. In 16% of samples, THg was above the limits set by the National Health Surveillance Agency (ANVISA, namely 1µgg-1, and 62% exceeded the World Health Organization (WHO limit of 0.5µgg-1. The ingestion of 100g/ day-1 of P. glauca or X. gladius would result in a daily THg intake of more than twice the WHO (1990 suggested limit.

  16. Rapid production of trees. [Acer platanoides, Fraxinus excelsior, Quercus robus, Sorbus, Picea, and Abies spp. , Betula verrucose, Populus trichocarpa

    Energy Technology Data Exchange (ETDEWEB)

    Haarjorg, A.

    1976-01-01

    Seedlings of Acer platanoides approximately 2 m tall were produced in southern Norway in one year by seed stratification indoors. Similar results were obtained with Fraxinus excelsior, Quercus robus and Sorbus spp. Trails were also carried out with Betula verrucose (B. pendula), Populus trichocarpa, Picea spp., Abies spp., and other conifers. In all trials growth was increased when plants were raised in a plastic house, and depended on the time that Spring growth was started or whether supplementary light was given and also depended on the seed strain. For northern and high altitude strains it was important to maintain critical day length.

  17. Effect of high dose SO2 and ethylene exposure on the structure of epicuticular wax of picea pungens

    International Nuclear Information System (INIS)

    Patrie, J.; Berg, V.

    1994-01-01

    Conifers in polluted air generally exhibit accelerated degradation of epicuticular wax, but it is not clear whether the change is due to direct exposure to the pollutant or some other mechanism. Needles from blue spruce (Picea pungens) were exposed to sulfur dioxide or ethylene gas at 0 to 10,000 microliters per liter for 2 to 196 h; samples were examined by scanning electron microscopy. Neither gas caused changes in the wax crystals, although late in the growing season a fungal infestation was associated with degradation of wax structures. This supports hypotheses explaining accelerated epicuticular wax degradation by indirect effects of exposure to air pollutants. (orig.)

  18. Multielement analysis of Picea rubens Sarg. Three-rings by proton-induced x-ray emission

    International Nuclear Information System (INIS)

    Hall, G.S.

    1984-01-01

    A 3.7 MeV external proton beam was employed to produce X-rays in Picea rubens Sarg. (red spruce). Intra-annual growth rings and several elements were quantitatively determined (P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Pb, Sr). The PIXE experimental set-up, rapid sample preparation methods, and some interesting observations are discussed. In addition, statistical analysis of the tree-ring element concentrations revealed that K, Cl, and Ca were slightly translocated whereas the other elements detected were not. 15 references, 1 figure, 1 table

  19. Total Stem and Merchantable Volume Equations of Norway Spruce (Picea abies (L.) Karst.) Growing on Former Farmland in Sweden

    OpenAIRE

    Johansson, Tord

    2014-01-01

    An equation was constructed to estimate the stem volume of Norway spruce (Picea abies (L.) Karst.) in 145 stands growing on former farmland in Sweden (Latitude 56-63 degrees N). The mean total age was 40 +/- 13 (range 17-91) years, the mean diameter at breast height (ob) was 15 +/- 4 (range 5-27) cm and the mean density was 1621 +/- 902 (range 100-7600) stems ha(-1). The equation which fits the data best used the diameter at breast height and total stem height as predictive variables. Merchan...

  20. Primeiro registro de Fidicinoides picea (Walker, 1850 e Fidicinoides poulaini Boulard & Martinelli, 1996 no Brasil First report of Fidicinoides picea (Walker, 1850 and Fidicinoides poulaini Boulard & Martinelli, 1996 in Brazil

    Directory of Open Access Journals (Sweden)

    Rodrigo Souza Santos

    2009-04-01

    Full Text Available As cigarras são importantes pragas de diversas culturas, entre plantas cultivadas e ornamentais. Estudos de taxonomia e morfologia de insetos são primordiais para pesquisas etológicas e para o emprego de métodos de controle. Ao estudar coleções entomológicas de cigarras, foi constatado o primeiro registro de ocorrência de Fidicinoides picea e Fidicinoides poulaini (Hemiptera: Cicadidae no Brasil, espécies coletadas nos Estados do Pará e de Manaus, respectivamente. Neste trabalho, são apresentadas as descrições morfológicas da cabeça, do tórax, do abdome e da asa.The cicadas are important pests of several cultivations, since cultivated until ornamental plants. Taxonomy and morphology studies of insects are primordial in researches of behavior and for the employment of control methods. Studying entomological collections of cicadas, it was verified the first report of occurrence of Fidicinoides picea and Fidicinoides poulaini (Hemiptera: Cicadidae in Brazil, species collected in States of Pará and Manaus and are presented, morphologic descriptions of the head, thorax, abdomen and wing.

  1. Final cutting of shelterwood. Harvesting techniques and effects on the Picea abies regeneration

    International Nuclear Information System (INIS)

    Gloede, Dan

    2001-01-01

    During the last decade, environmental and biological aspects have grown increasingly important in forestry. At the same time conventional planting after clear-cutting has failed on many sites with a high ground water table, abundant competitive vegetation and frequent frosts. Therefore, on these sites the use of the shelterwood system for regeneration of Norway spruce (Picea abies [L.] Karst.) has increased in Sweden. The main objective of the thesis is to study if it is possible to final-cut shelterwoods at acceptable harvesting costs, logging damage and release effects in the regeneration. Final cutting of three shelterwoods (180-200 m 3 /ha) in Sweden were carried out with single- and double-grip harvester systems in 1-1.5 m high regeneration (6 400-26 700 seedlings/ha). In a fourth shelterwood (140-165 m 3 /ha), also situated in Sweden, conventional felling with a single-grip harvester was compared with a more concentrated felling according to a method named 'tossing the caber', where the trees were felled top-end first over the 1.2-1.3 m high regeneration (9 530-11 780 seedlings/ha) and into the striproad. No differences in productivity and cost between single- and double-grip harvesters in final cutting of shelterwood were found. Despite few stems/ha and extensive regeneration the harvesting cost was considered low (33.5 SEK/m 3 ). Approximately one third of the seedlings suffered mortal logging damage, which was considered acceptable. No differences between conventional felling and the tossing the caber method were found regarding productivity, cost and damage to the regeneration. However, tossing the caber may be a more productive alternative in final cutting of pine-dominated shelterwood or seed tree stands. Seedling growth and survival after shelterwood removal was not influenced by the choice of harvester system. Seedling height and vitality were found to be good estimators of post-release survival and growth which, in total, was found to be acceptable

  2. Construction of a nurse shark (Ginglymostoma cirratum) bacterial artificial chromosome (BAC) library and a preliminary genome survey

    OpenAIRE

    Luo, Meizhong; Kim, HyeRan; Kudrna, Dave; Sisneros, Nicholas B; Lee, So-Jeong; Mueller, Christopher; Collura, Kristi; Zuccolo, Andrea; Buckingham, E Bryan; Grim, Suzanne M; Yanagiya, Kazuyo; Inoko, Hidetoshi; Shiina, Takashi; Flajnik, Martin F; Wing, Rod A

    2006-01-01

    Abstract Background Sharks are members of the taxonomic class Chondrichthyes, the oldest living jawed vertebrates. Genomic studies of this group, in comparison to representative species in other vertebrate taxa, will allow us to theorize about the fundamental genetic, developmental, and functional characteristics in the common ancestor of all jawed vertebrates. Aims In order to obtain mapping and sequencing data for comparative genomics, we constructed a bacterial artificial chromosome (BAC) ...

  3. Generating resources for genomics of wheat homoeologous chromosome group 3: 3AS- and 3DS-specific BAC libraries

    Czech Academy of Sciences Publication Activity Database

    Šafář, Jan; Šimková, Hana; Kubaláková, Marie; Suchánková, Pavla; Čihalíková, Jarmila; Bartoš, Jan; Fiocchetti, F.; Roselli, M.; Gill, B. S.; Doležel, Jaroslav; Lucretti, S.

    2009-01-01

    Roč. 61, 1-2 (2009), s. 151-160 ISSN 0394-9257 R&D Projects: GA ČR GD521/05/H013; GA ČR GA521/06/1723; GA ČR GA521/07/1573; GA MŠk(CZ) LC06004; GA MŠk OC08025 Institutional research plan: CEZ:AV0Z50380511 Keywords : BAC library * Flow sorting * Homoeologous chromosomes Subject RIV: GE - Plant Breeding

  4. PLE-wu, a new member of piggyBac transposon family from insect, is active in mammalian cells.

    Science.gov (United States)

    Wu, Chunxiao; Wang, Shu

    2014-10-01

    piggyBac, a highly active transposon in insect and mammalian cells, is a very useful tool in genome manipulation. A new piggyBac-like element (PLE), named PLE-wu, was identified from a mutant baculovirus cultured in sf9 insect cells. This new transposon is 2931 bp in length and encodes two active forms of transposase, a 708-amino acid-long transposase and a short 576-residue-long transposase translated from a downstream in-frame initiation codon. PLE-wu has asymmetric terminal structures, containing 6-bp inverted terminal repeats, 32-bp imperfect inverted and direct sub-terminal repeats. Similar to piggyBac, PLE-wu exhibits traceless excision activity in both insect and mammalian cells, restoring the original TTAA target sequence upon excision. It also retains the insertion activity in mammalian cells with a plasmid to chromosome transposition rate about 10-fold higher than random integration. Plasmid rescue assays revealed that the TTAA target sequence was duplicated at the junctions of the insertion site. Deletion of the terminal sequences including the sub-terminal repeats decreased the transposition activity of the 708-residue-long transposase, while the transposition activity of the short form of transposase was not affected. With its low sequence similarity to piggyBac, PLE-wu will contribute to the understanding the mechanism of PLE transposition, as well as design of new transposon systems with higher activity. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  5. BAC-derived markers converted from RFLP linked to Phytophthora capsici resistance in pepper (Capsicum annuum L.).

    Science.gov (United States)

    Kim, Hyoun-Joung; Nahm, Seok-Hyeon; Lee, Heung-Ryul; Yoon, Gi-Bo; Kim, Ki-Taek; Kang, Byoung-Cheorl; Choi, Doil; Kweon, Oh Yeol; Cho, Myeong-Cheoul; Kwon, Jin-Kyung; Han, Jung-Heon; Kim, Jeong-Ho; Park, Minkyu; Ahn, Jong Hwa; Choi, Soon Ho; Her, Nam Han; Sung, Joo-Hee; Kim, Byung-Dong

    2008-12-01

    Phytophthora capsici Leonian, an oomycete pathogen, is a serious problem in pepper worldwide. Its resistance in pepper is controlled by quantitative trait loci (QTL). To detect QTL associated with P. capsici resistance, a molecular linkage map was constructed using 100 F(2) individuals from a cross between Capsicum annuum 'CM334' and C. annuum 'Chilsungcho'. This linkage map consisted of 202 restriction fragment length polymorphisms (RFLPs), 6 WRKYs and 1 simple sequence repeat (SSR) covering 1482.3 cM, with an average interval marker distance of 7.09 cM. QTL mapping of Phytophthora root rot and damping-off resistance was performed in F(2:3) originated from a cross between resistant Mexican landrace C. annuum 'CM334' and susceptible Korean landrace C. annuum 'Chilsungcho' using composite interval mapping (CIM) analysis. Four QTL explained 66.3% of the total phenotypic variations for root rot resistance and three 44.9% for damping-off resistance. Of these QTL loci, two were located close to RFLP markers CDI25 on chromosome 5 (P5) and CT211A on P9. A bacterial artificial chromosome (BAC) library from C. annuum 'CM334' was screened with these two RFLP probes to obtain sequence information around the RFLP marker loci for development of PCR-based markers. CDI25 and CT211 probes identified seven and eight BAC clones, respectively. Nine positive BAC clones containing probe regions were sequenced and used for cytogenetic analysis. One single-nucleotide amplified polymorphism (SNAP) for the CDI25 locus, and two SSRs and cleaved amplified polymorphic sequence (CAPS) for CT211 were developed using sequences of the positive BAC clones. These markers will be valuable for rapid selection of genotypes and map-based cloning for resistance genes against P. capsici.

  6. Driving performance on the descending limb of blood alcohol concentration (BAC) in undergraduate students: a pilot study.

    Science.gov (United States)

    Tremblay, Mathieu; Gallant, François; Lavallière, Martin; Chiasson, Martine; Silvey, Dustin; Behm, David; Albert, Wayne J; Johnson, Michel J

    2015-01-01

    Young drivers are overrepresented in collisions resulting in fatalities. It is not uncommon for young drivers to socially binge drink and decide to drive a vehicle a few hours after consumption. To better understand the risks that may be associated with this behaviour, the present study has examined the effects of a social drinking bout followed by a simulated drive in undergraduate students on the descending limb of their BAC (blood alcohol concentration) curve. Two groups of eight undergraduate students (n = 16) took part in this study. Participants in the alcohol group were assessed before drinking, then at moderate and low BAC as well as 24 hours post-acute consumption. This group consumed an average of 5.3 ± 1.4 (mean ± SD) drinks in an hour in a social context and were then submitted to a driving and a predicted crash risk assessment. The control group was assessed at the same time points without alcohol intake or social context.; at 8 a.m., noon, 3 p.m. and 8 a.m. the next morning. These multiple time points were used to measure any potential learning effects from the assessment tools (i.e. driving simulator and useful field of view test (UFOV)). Diminished driving performance at moderate BAC was observed with no increases in predicted crash risk. Moderate correlations between driving variables were observed. No association exists between driving variables and UFOV variables. The control group improved measures of selective attention after the third assessment. No learning effect was observed from multiple sessions with the driving simulator. Our results show that a moderate BAC, although legal, increases the risky behaviour. Effects of alcohol expectancy could have been displayed by the experimental group. UFOV measures and predicted crash risk categories were not sensitive enough to predict crash risk for young drivers, even when intoxicated.

  7. Association of ectomycorrhizal fungi with Picea crassifolia (Pinaceae, Piceoidae) from high-altitude stands in Mount Helan Nature Reserve, China.

    Science.gov (United States)

    Fan, Y J; Grebenc, T; Wei, J; Zhao, Y L; Yan, W; Wang, L B

    2016-09-02

    We investigated the diversity of ectomycorrhiza associated with the endemic Picea crassifolia in Mount Helan National Nature Reserve in Inner Mongolia, China. Toward this objective, we conducted morphological and molecular identification of ectomycorrhizae in soil cubes taken from pure P. crassifolia stands. Eleven types of ectomycorrhizal (ECM) organisms were separated, briefly described, and identified. Nine morphotypes belonged to the phylum Basidiomycotina [Amphinema byssoides, Cortinarius sp (cf. limonius), Cortinarius vernus, Inocybe cf. nitidiscula, Inocybe sp 1, Sebacina incrustans, Sebacina sp, Suillus luteus, and Piceirhiza tuberculata x Picea crassifolia (comb. Nov.)], and two morphotypes to the phylum Ascomycotina (Cenococcum geophilum and Helvella sp). The diversity of ECM organisms in P. crassifolia was lower than that reported by other studies on spruce or pine forests, or on sporocarp diversity in the high-mountain forests of China. Most of the fungi in the rhizosphere did not correspond to species previously recorded as sporocarps above ground. Here, several new ectomycorrhiza morphotypes are proposed and described. We also confirmed the ectomycorrhizal status of the genus Sebacina (order Sebacinales).

  8. Evaluation of chronic lymphocytic leukemia by BAC-based microarray analysis

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    McDaniel Lisa D

    2011-02-01

    Full Text Available Abstract Background Chronic lymphocytic leukemia (CLL is a highly variable disease with life expectancies ranging from months to decades. Cytogenetic findings play an integral role in defining the prognostic significance and treatment for individual patients. Results We have evaluated 25 clinical cases from a tertiary cancer center that have an established diagnosis of CLL and for which there was prior cytogenetic and/or fluorescence in situ hybridization (FISH data. We performed microarray-based comparative genomic hybridization (aCGH using a bacterial artificial chromosome (BAC-based microarray designed for the detection of known constitutional genetic syndromes. In 15 of the 25 cases, aCGH detected all copy number imbalances identified by prior cytogenetic and/or FISH studies. For the majority of those not detected, the aberrations were present at low levels of mosaicism. Furthermore, for 15 of the 25 cases, additional abnormalities were detected. Four of those cases had deletions that mapped to intervals implicated in inherited predisposition to CLL. For most cases, aCGH was able to detect abnormalities present in as few as 10% of cells. Although changes in ploidy are not easily discernable by aCGH, results for two cases illustrate the detection of additional copy gains and losses present within a mosaic tetraploid cell population. Conclusions Our results illustrate the successful evaluation of CLL using a microarray optimized for the interrogation of inherited disorders and the identification of alterations with possible relevance to CLL susceptibility.

  9. Vector modifications to eliminate transposase expression following piggyBac-mediated transgenesis

    Science.gov (United States)

    Chakraborty, Syandan; Ji, HaYeun; Chen, Jack; Gersbach, Charles A.; Leong, Kam W.

    2014-01-01

    Transgene insertion plays an important role in gene therapy and in biological studies. Transposon-based systems that integrate transgenes by transposase-catalyzed “cut-and-paste” mechanism have emerged as an attractive system for transgenesis. Hyperactive piggyBac transposon is particularly promising due to its ability to integrate large transgenes with high efficiency. However, prolonged expression of transposase can become a potential source of genotoxic effects due to uncontrolled transposition of the integrated transgene from one chromosomal locus to another. In this study we propose a vector design to decrease post-transposition expression of transposase and to eliminate the cells that have residual transposase expression. We design a single plasmid construct that combines the transposase and the transpositioning transgene element to share a single polyA sequence for termination. Consequently, the separation of the transposase element from the polyA sequence after transposition leads to its deactivation. We also co-express Herpes Simplex Virus thymidine kinase (HSV-tk) with the transposase. Therefore, cells having residual transposase expression can be eliminated by the administration of ganciclovir. We demonstrate the utility of this combination transposon system by integrating and expressing a model therapeutic gene, human coagulation Factor IX, in HEK293T cells. PMID:25492703

  10. Compare the influence of several methods dehydrate of cajuput (Melaleuca cajuputi) honey from Bac Lieu - Vietnam

    Science.gov (United States)

    Nam, Nguyen Xuan; Phuc, Nguyen Chi; Oanh, Huynh Ngoc; Hien, Phan Phuoc

    2017-09-01

    The aim of this research was exhaustive to valuate the effects of 5 methods used to dehydrate of cajuput honey from Bac Lieu - Vietnam include thermal, microwave, ultrasonic wave, silicagel, vacuum processing on water content of honey. Beside that, comparing the effects on honey quality with the industrial-scale processing. The main honey quality paramenters (reducing sugar (RS), hydroxymethylfurfural (HMF), diastase number (DN), water and colour) of cajuput honey were analyzed. RS content were analyzed by DNS method, HMF under AOAC 980.23, diastase activity based on AOAC 958.09, water content according to AOAC 969.38B and colour parameters (L*, a*, b*) were established in the CIE system. The physico-chemical characteristics of fresh honey was as follows: water content 23.18%, RS 717.42 mg/g, HMF 4.24 mg/kg, DN 4.85 mg/kg, colour parameters L*a*b* 39.51-10.51-31.81. The results of the analysis showed that excepts ultrasonic wave processing (22.93%), the other methods allowed to dehydrate of cajuput honey (thermal - 18.73%, microwave - 16.87%, silicagel - 17.86%, vacuum - 17.35% and industrial-scale processing - 16.85%).

  11. Identification of biofilm-associated cluster (bac in Pseudomonas aeruginosa involved in biofilm formation and virulence.

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    Camille Macé

    Full Text Available Biofilms are prevalent in diseases caused by Pseudomonas aeruginosa, an opportunistic and nosocomial pathogen. By a proteomic approach, we previously identified a hypothetical protein of P. aeruginosa (coded by the gene pA3731 that was accumulated by biofilm cells. We report here that a Delta pA3731 mutant is highly biofilm-defective as compared with the wild-type strain. Using a mouse model of lung infection, we show that the mutation also induces a defect in bacterial growth during the acute phase of infection and an attenuation of the virulence. The pA3731 gene is found to control positively the ability to swarm and to produce extracellular rhamnolipids, and belongs to a cluster of 4 genes (pA3729-pA3732 not previously described in P. aeruginosa. Though the protein PA3731 has a predicted secondary structure similar to that of the Phage Shock Protein, some obvious differences are observed compared to already described psp systems, e.g., this unknown cluster is monocistronic and no homology is found between the other proteins constituting this locus and psp proteins. As E. coli PspA, the amount of the protein PA3731 is enlarged by an osmotic shock, however, not affected by a heat shock. We consequently named this locus bac for biofilm-associated cluster.

  12. Germ line transformation of the yellow fever mosquito, Aedes aegypti, mediated by transpositional insertion of a piggyBac vector.

    Science.gov (United States)

    Lobo, N F; Hua-Van, A; Li, X; Nolen, B M; Fraser, M J

    2002-04-01

    Mosquito-vectored diseases such as yellow fever and dengue fever continue to have a substantial impact on human populations world-wide. Novel strategies for control of these mosquito vectored diseases can arise through the development of reliable systems for genetic manipulation of the insect vector. A piggyBac vector marked with the Drosophila melanogaster cinnabar (cn) gene was used to transform the white-eyed khw strain of Aedes aegypti. Microinjection of preblastoderm embryos resulted in four families of cinnabar transformed insects. An overall transformation frequency of 4%, with a range of 0% to as high as 13% for individual experiments, was achieved when using a heat-shock induced transposase providing helper plasmid. Southern hybridizations indicated multiple insertion events in three of four transgenic lines, while the presence of duplicated target TTAA sites at either ends of individual insertions confirmed characteristic piggyBac transposition events in these three transgenic lines. The transgenic phenotype has remained stable for more than twenty generations. The transformations effected using the piggyBac element establish the potential of this element as a germ-line transformation vector for Aedine mosquitoes.

  13. Construction of a 7-fold BAC library and cytogenetic mapping of 10 genes in the giant panda (Ailuropoda melanoleuca

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    Zhang Ying

    2006-11-01

    Full Text Available Abstract Background The giant panda, one of the most primitive carnivores, is an endangered animal. Although it has been the subject of many interesting studies during recent years, little is known about its genome. In order to promote research on this genome, a bacterial artificial chromosome (BAC library of the giant panda was constructed in this study. Results This BAC library contains 198,844 clones with an average insert size of 108 kb, which represents approximately seven equivalents of the giant panda haploid genome. Screening the library with 15 genes and 8 microsatellite markers demonstrates that it is representative and has good genome coverage. Furthermore, ten BAC clones harbouring AGXT, GHR, FSHR, IRBP, SOX14, TTR, BDNF, NT-4, LH and ZFX1 were mapped to 8 pairs of giant panda chromosomes by fluorescence in situ hybridization (FISH. Conclusion This is the first large-insert genomic DNA library for the giant panda, and will contribute to understanding this endangered species in the areas of genome sequencing, physical mapping, gene cloning and comparative genomic studies. We also identified the physical locations of ten genes on their relative chromosomes by FISH, providing a preliminary framework for further development of a high resolution cytogenetic map of the giant panda.

  14. The host antimicrobial peptide Bac71-35 binds to bacterial ribosomal proteins and inhibits protein synthesis.

    Science.gov (United States)

    Mardirossian, Mario; Grzela, Renata; Giglione, Carmela; Meinnel, Thierry; Gennaro, Renato; Mergaert, Peter; Scocchi, Marco

    2014-12-18

    Antimicrobial peptides (AMPs) are molecules from innate immunity with high potential as novel anti-infective agents. Most of them inactivate bacteria through pore formation or membrane barrier disruption, but others cross the membrane without damages and act inside the cells, affecting vital processes. However, little is known about their intracellular bacterial targets. Here we report that Bac71-35, a proline-rich AMP belonging to the cathelicidin family, can reach high concentrations (up to 340 μM) inside the E. coli cytoplasm. The peptide specifically and completely inhibits in vitro translation in the micromolar concentration range. Experiments of incorporation of radioactive precursors in macromolecules with E. coli cells confirmed that Bac71-35 affects specifically protein synthesis. Ribosome coprecipitation and crosslinking assays showed that the peptide interacts with ribosomes, binding to a limited subset of ribosomal proteins. Overall, these results indicate that the killing mechanism of Bac71-35 is based on a specific block of protein synthesis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Crystal structures and electronic properties of BaC2 isomers by theoretical study based on DFT

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Band structures and electronic properties of two BaC2 isomers were calculated by using density func-tional theory(DFT) properly.The ionic bond features are all typical between cation(Ba) and anion clusters(C2) in both structures of the isomers.However,a much stronger covalent bond exists in anion clusters which can be seen by inspecting the electron distribution contour that has a dull bell like shape between two carbon atoms.The shortest distance between Ba2+ and C22? and the bond length in anion clusters are different in these isomers of BaC2,which are 0.2945 nm and 0.1185 nm for the structure with the I4/mmm space group and 0.2744 and 0.1136 nm with the C2/c type,respectively.Band structures were clarified by combining the DOS to indicate the ionic bonding features more clearly.Population analysis provided further evidence on these ideas.Thermodynamical calculation results reveal that the transition temperature of these two polymorphs of BaC2 locates near 132 K,which is consistent with the recent experimental results.

  16. A nucleolus-predominant piggyBac transposase, NP-mPB, mediates elevated transposition efficiency in mammalian cells.

    Science.gov (United States)

    Hong, Jin-Bon; Chou, Fu-Ju; Ku, Amy T; Fan, Hsiang-Hsuan; Lee, Tung-Lung; Huang, Yung-Hsin; Yang, Tsung-Lin; Su, I-Chang; Yu, I-Shing; Lin, Shu-Wha; Chien, Chung-Liang; Ho, Hong-Nerng; Chen, You-Tzung

    2014-01-01

    PiggyBac is a prevalent transposon system used to deliver transgenes and functionally explore the mammalian untouched genomic territory. The important features of piggyBac transposon are the relatively low insertion site preference and the ability of seamless removal from genome, which allow its potential uses in functional genomics and regenerative medicine. Efforts to increase its transposition efficiency in mammals were made through engineering the corresponding transposase (PBase) codon usage to enhance its expression level and through screening for mutant PBase variants with increased enzyme activity. To improve the safety for its potential use in regenerative medicine applications, site-specific transposition was achieved by using engineered zinc finger- and Gal4-fused PBases. An excision-prone PBase variant has also been successfully developed. Here we describe the construction of a nucleolus-predominant PBase, NP-mPB, by adding a nucleolus-predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon-optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nucleolar organizer regions is not detected. Instead a 3-4 fold increase in piggyBac transposition efficiency is reproducibly observed in mouse and human cells.

  17. Ozone fumigation under dark/light conditions of Norway Spruce (Picea Abies) and Scots Pine (Pinus Sylvestris)

    Science.gov (United States)

    Canaval, Eva; Jud, Werner; Hansel, Armin

    2015-04-01

    Norway Spruce (Picea abies) and Scots Pine (Pinus sylvestris) represent dominating tree species in the northern hemisphere. Thus, the understanding of their ozone sensitivity in the light of the expected increasing ozone levels in the future is of great importance. In our experiments we investigated the emissions of volatile organic compounds (VOCs) of 3-4 year old Norway Spruce and Scots Pine seedlings under ozone fumigation (50-150 ppbv) and dark/light conditions. For the experiments the plants were placed in a setup with inert materials including a glass cuvette equipped with a turbulent air inlet and sensors for monitoring a large range of meteorological parameters. Typical conditions were 20-25°C and a relative humidity of 70-90 % for both plant species. A fast gas exchange rate was used to minimize reactions of ozone in the gas phase. A Switchable-Reagent-Ion-Time-of-Flight-MS (SRI-ToF-MS) was used to analyze the VOCs at the cuvette outlet in real-time during changing ozone and light levels. The use of H3O+ and NO+ as reagent ions allows the separation of certain isomers (e.g. aldehydes and ketones) due to different reaction pathways depending on the functional groups of the molecules. Within the Picea abies experiments the ozone loss, defined as the difference of the ozone concentration between cuvette inlet and outlet, remained nearly constant at the transition from dark to light. This indicates that a major part of the supplied ozone is depleted non-stomatally. In contrast the ozone loss increased by 50 % at the transition from dark to light conditions within Pinus sylvestris experiments. In this case the stomata represent the dominant loss channel. Since maximally 0.1% of the ozone loss could be explained by gas phase reactions with monoterpenes and sesquiterpenes, we suggest that ozone reactions on the surface of Picea abies represent the major sink in this case and lead to an light-independent ozone loss. This is supported by the fact that we detected

  18. Sur la filabilité d'une laine teinte en bac-ouvert et d'une laine teinte à la continue

    NARCIS (Netherlands)

    Werker, W.; Mulder, D.; Stomph, J.; Schartman, A.F.

    1966-01-01

    Afin de constater, si la teinture selon le procédé à la continue ou selon le procédé en bac-ouvert influence les résultats dans Ia filature, on a teint deux lots homogènes à 18OO kg par lot en quatre parties: 450 kg, procédé à la continue, teints en rouge, 450 kg, procédé bac-ouvert, teints en

  19. Experimental tolerance to boron of the plant species Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea in Argentina; Tolerancia experimental de las especies vegetales Nicotiana glauca, Jacaranda mimosifolia, Tecoma stans, Medicago sativa y Spinacea oleracea al boro, en Argentina

    Energy Technology Data Exchange (ETDEWEB)

    Viana, Marta L. de; Albarracin Franco, Silvia [Univ. Nacional de Salta, Inst. de Ecologia y Ambiente Humano, CIUNSa, Buenos Aires No. 177, 4400, Salta (Argentina)], E-mail: mldeviana@arnet.com.ar

    2008-09-15

    The activity of the borate deposits industries constitutes a point source and diffuse pollution of air, soil and water. Therefore, the study and experimentation on possible ways to offset this impact is a priority. A relatively new technique to decontaminate soils is phytoremediation, which uses plants and associated microorganisms. The first step is to identify tolerant plant species, which is the focus of this work. An experiment was conducted in the laboratory to evaluate the germination, survival and growth of different species in different concentrations of boron. At the beginning and end of the experiment was determined concentration of boron in the substrate for each treatment and for substrates with and without vegetation. Significant differences due to treatment, the species and species-treatment interaction. M. sativa, N. glauca and J. mimosifolia were the species most tolerant to boron. The other species showed a decrease in all variables-response function of the concentration of the contaminant. All had low survival in the highest concentration. The decrease of boron was highest in the treatment of 30 ppm of boron with M. sativa and the lowest was recorded in the treatment of 20 ppm of boron with J. mimosifolia and 30 ppm of boron with T. stans and S. oleracea. It is concluded that N. glauca, M. sativa and J. mimosifolia could be considered as promising remediation. (author) [Spanish] La actividad de las industrias borateras constituye una fuente puntual y difusa de contaminacion del aire, suelo y aguas superficiales y profundas. Por lo tanto, el estudio y experimentacion acerca de las posibles formas de contrarrestar este impacto constituye una prioridad. Una tecnica relativamente nueva para descontaminar suelos es la fitorremediacion, que emplea plantas y microorganismos asociados. El primer paso es detectar las especies vegetales tolerantes, lo que constituye el objetivo de este trabajo. Se realizo un experimento en laboratorio para evaluar la

  20. Isolation of a 97-kb minimal essential MHC B locus from a new reverse-4D BAC library of the golden pheasant.

    Directory of Open Access Journals (Sweden)

    Qing Ye

    Full Text Available The bacterial artificial chromosome (BAC system is widely used in isolation of large genomic fragments of interest. Construction of a routine BAC library requires several months for picking clones and arraying BACs into superpools in order to employ 4D-PCR to screen positive BACs, which might be time-consuming and laborious. The major histocompatibility complex (MHC is a cluster of genes involved in the vertebrate immune system, and the classical avian MHC-B locus is a minimal essential one, occupying a 100-kb genomic region. In this study, we constructed a more effective reverse-4D BAC library for the golden pheasant, which first creates sub-libraries and then only picks clones of positive sub-libraries, and identified several MHC clones within thirty days. The full sequencing of a 97-kb reverse-4D BAC demonstrated that the golden pheasant MHC-B locus contained 20 genes and showed good synteny with that of the chicken. The notable differences between these two species were the numbers of class II B loci and NK genes and the inversions of the TAPBP gene and the TAP1-TAP2 region. Furthermore, the inverse TAP2-TAP1 was unique in the golden pheasant in comparison with that of chicken, turkey, and quail. The newly defined genomic structure of the golden pheasant MHC will give an insight into the evolutionary history of the avian MHC.

  1. The canine sarcoglycan delta gene: BAC clone contig assembly, chromosome assignment and interrogation as a candidate gene for dilated cardiomyopathy in Dobermann dogs.

    Science.gov (United States)

    Stabej, P; Leegwater, P A J; Imholz, S; Versteeg, S A; Zijlstra, C; Stokhof, A A; Domanjko-Petriè, A; van Oost, B A

    2005-01-01

    Dilated cardiomyopathy (DCM) is a common disease of the myocardium recognized in human, dog and experimental animals. Genetic factors are responsible for a large proportion of cases in humans, and 17 genes with DCM causing mutations have been identified. The genetic origin of DCM in the Dobermann dogs has been suggested, but no disease genes have been identified to date. In this paper, we describe the characterization and evaluation of the canine sarcoglycan delta (SGCD), a gene implicated in DCM in human and hamster. Bacterial artificial chromosomes (BACs) containing the canine SGCD gene were isolated with probes for exon 3 and exons 4-8 and were characterized by Southern blot analysis. BAC end sequences were obtained for four BACs. Three of the BACs overlapped and could be ordered relative to each other and the end sequences of all four BACs could be anchored on the preliminary assembly of the dog genome sequence (www. ensembl.org). One of the BACs of the partial contig was localized by fluorescent in situ hybridization to canine chromosome 4q22, in agreement with the dog genome sequence. Two highly informative polymorphic microsatellite markers in intron 7 of the SGCD gene were identified. In 25 DCM-affected and 13 non DCM-affected dogs seven different haplotypes could be distinguished. However, no association between any of the SGCD variants and the disease locus was apparent.

  2. Development and microstructure of tooth histotypes in the blue shark, Prionace glauca (Carcharhiniformes: Carcharhinidae) and the great white shark, Carcharodon carcharias (Lamniformes: Lamnidae).

    Science.gov (United States)

    Moyer, Joshua K; Riccio, Mark L; Bemis, William E

    2015-07-01

    Elasmobranchs exhibit two distinct arrangements of mineralized tissues in the teeth that are known as orthodont and osteodont histotypes. Traditionally, it has been said that orthodont teeth maintain a pulp cavity throughout tooth development whereas osteodont teeth are filled with osteodentine and lack a pulp cavity when fully developed. We used light microscopy, scanning electron microscopy, and high-resolution micro-computed tomography to compare the structure and development of elasmobranch teeth representing the two histotypes. As an example of the orthodont histotype, we studied teeth of the blue shark, Prionace glauca (Carcharhiniformes: Carcharhinidae). For the osteodont histotype, we studied teeth of the great white shark, Carcharodon carcharias (Lamniformes: Lamnidae). We document similarities and differences in tooth development and the microstructure of tissues in these two species and review the history of definitions and interpretations of elasmobranch tooth histotypes. We discuss a possible correlation between tooth histotype and tooth replacement and review the history of histotype differentiation in sharks. We find that contrary to a long held misconception, there is no orthodentine in the osteodont teeth of C. carcharias. © 2015 Wiley Periodicals, Inc.

  3. Effects of Harvesting Systems and Bole Moisture Loss on Weight Scaling of Douglas-Fir Sawlogs (Pseudotsuga Menziesii var. glauca Franco

    Directory of Open Access Journals (Sweden)

    Jarred D. Saralecos

    2014-09-01

    Full Text Available Characterizing the moisture loss from felled trees is essential for determining weight-to-volume (W-V relationships in softwood sawlogs. Several factors affect moisture loss, but research to quantify the effects of bole size and harvest method is limited. This study was designed to test whether bole size, harvest method, environmental factors, and the associated changes in stem moisture content of felled Douglas-fir (Pseudotsuga menziesii var. glauca Franco affected the weight-to-volume relationship of sawlogs. Thirty trees in three size classes (12.7–25.4 cm, 25.5–38.1 cm, 38.2–50.8 cm were felled and treated with one of two harvesting processing methods. Moisture content was sampled every two days for four weeks. Results showed 6% greater moisture loss in the crowns of stems that retained limbs after felling compared to stems with limbs removed after harvesting. Additionally, moisture loss rate increased as stem size decreased. The smallest size class lost 58% moisture content compared to 34% in the largest size class throughout the study duration. These stem moisture content changes showed a 17% reduction in average sawlog weight within the largest size class, shifting current W-V relationships from 2.33 tons m−3 to 1.94 tons m−3 during the third seasonal quarter for northern Idaho Douglas-fir and potentially altering relationships year-round.

  4. ALCALOIDES BISBENCILISOQUINOLINICOS AISLADOS DE LA ESPECIE NATIVA Berberís glauca (HBK Y EVALUACIÓN DE SU ACTIVIDAD ANTIALIMENTARIA

    Directory of Open Access Journals (Sweden)

    Barbara Moreno Murillo

    2010-09-01

    Full Text Available Durante el estudio de la potencial actividad tóxica del extracto alcohólico de ios tallos de la especie nativa Berberís glauca (HBK (Berberidaceae. Se realizaron bioensayos con larvas del crustáceo Artemia salina, y de los insectos Musca domestica (Díptera y Spodoptera sunia (Lepidoptera. La fracción alcaloidal soluble en CHCl, obtenida por extracción ácido-base del extracto etanólico mostró notable acción antialimentaria frente a larvas de 3° y 4° estadio de la polilla S. sunia. La separación cromatográfica guiada por bioensayos permitió aislar y purificar de la fracción de mayor actividad varios alcaloides benciltctrahidroisoquinolínicos. Por métodos espectroscópicos se identificaron berbamina, isotalicberina. O- metilisotalicberina y 7- 0-demetilisotalicberina. Este es el primer reporte sobre la presencia de bases diméricas bisbenciltetrahidroisoquinolínicas (bbi en especies de Berberís colombianas. Además se detectó la presencia de una base dimérica novedosa de PM 712 cuya dilucidación estructural no se ha establecido en forma completa.

  5. Alcaloides bisbencilisoquinolinicos aislados de la especie nativa berberís glauca (HBK y evaluación de su actividad antialimentaria

    Directory of Open Access Journals (Sweden)

    Bárbara Moreno-Murillo

    2010-07-01

    Full Text Available Durante el estudio de la potencial actividad tóxica del extracto alcohólico de ios tallos de la especie nativa Berberís glauca (HBK (Berberidaceae. se realizaron bioensayos con larvas del crustáceo Artemia salina, y de los insectos Musca domestica (Díptera y Spodoptera sunia (Lepidoptera. La fracción alcaloidal soluble en CHCl, obtenida por extracción ácido-base del extracto etanólico mostró notable acción antialimentaria frente a larvas de 3° y 4° estadio de la polilla S. sunia. La separación cromatográfica guiada por bioensayos permitió aislar y purificar de la fracción de mayor actividad varios alcaloides benciltctrahidroisoquinolínicos. Por métodos espectroscópicos se identificaron berbamina, isotalicberina. O- metilisotalicberina y 7- 0-demetilisotalicberina. Este es el primer reporte sobre la presencia de bases diméricas bisbenciltetrahidroisoquinolínicas (bbi en especies de Berberís colombianas. Además se detectó la presencia de una base dimérica novedosa de PM 712 cuya dilucidación estructural no se ha establecido en forma completa.

  6. Functions for biomass and basic density of stem, crown and root system of Norway spruce (Picea abies (L.) Karst.) in Denmark

    DEFF Research Database (Denmark)

    Skovsgaard, Jens Peter; Bald, Caroline; Nord-Larsen, Thomas

    2011-01-01

    Models for predicting the biomass of forest trees are becoming increasingly important for assessing forest resources and carbon sequestration in forests. We developed functions for predicting the biomass and basic density of above- and below-ground parts of Norway spruce (Picea abies (L.) Karst.)...

  7. Migration and population expansion of Abies, Fagus, Picea, and Quercus since 15000 years in and across the Alps, based on pollen-percentage threshold values

    NARCIS (Netherlands)

    Knaap, W.O. van der; Leeuwen, J.F.N. van; Finsinger, W.; Gobet, E.; Pini, R.; Schweizer, A.; Valsecchi, V.; Ammann, B.

    2005-01-01

    Aims: The aim of this study is to explore the migration (colonization of new areas) and subsequent population expansion (within an area) since 15 ka cal BP of Abies, Fagus, Picea, and Quercus into and through the Alps solely on the basis of high-quality pollen data. Methods: Chronologies of 101

  8. Bacterial and abiotic decay in waterlogged archaeological Picea abies (L.) Karst studied by confocal Raman imaging and ATR-FTIR spectroscopy

    DEFF Research Database (Denmark)

    Pedersen, Nanna Bjerregaard; Gierlinger, Notburga; Thygesen, Lisbeth Garbrecht

    2015-01-01

    Waterlogged archaeological Norway spruce [Picea abies (L.) Karst] poles were studied by means of confocal Raman imaging (CRI) and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) analysis to determine lignin and polysaccharide composition and distribution in the cell......, and minor oxidation of the lignin polymer compared to recent reference material. This is evidence for abiotic decay in the course of waterlogging....

  9. A short and efficient enantioselective synthesis of (+) and (-)-(Z)-7,15-hexadecadien-4-olide. The sex pheromone of the yellowish elongate chafer, Heptophylla picea

    Energy Technology Data Exchange (ETDEWEB)

    Clososki, Giuliano C.; Ricci, Luis C.; Costa, Carlos E.; Comasseto, Joao V. [Sao Paulo Univ., SP (Brazil). Inst. de Quimica]. E-mail: jvcomass@iq.usp.br

    2004-12-01

    The (R) and the (S) enantiomers of the Z-7,15-hexadecadien-4-olide (4), the sex pheromone of Heptophylla picea, were synthesized. A known lipase-catalysed enantiolactonization in the key step afforded a common precursor for both enantiomers of the pheromone in 92% e.e. (author)

  10. A short and efficient enantioselective synthesis of (+) and (-)-(Z)-7,15-hexadecadien-4-olide. The sex pheromone of the yellowish elongate chafer, Heptophylla picea

    International Nuclear Information System (INIS)

    Clososki, Giuliano C.; Ricci, Luis C.; Costa, Carlos E.; Comasseto, Joao V.

    2004-01-01

    The (R) and the (S) enantiomers of the Z-7,15-hexadecadien-4-olide (4), the sex pheromone of Heptophylla picea, were synthesized. A known lipase-catalysed enantiolactonization in the key step afforded a common precursor for both enantiomers of the pheromone in 92% e.e. (author)

  11. CITOGENETICS EFFECTS INDUCED BY THE ADMINISTRATION OF SUBSTANCES WITH REDUCTION POTENTIAL AT LARIX DECIDUA MILL. SSP. CARPATICA, PICEA ABIES (L. KARST. AND THUJA ORIENTALIS L

    Directory of Open Access Journals (Sweden)

    Ioana Anca Ieremia

    2007-08-01

    Full Text Available The paper present the influence of ascorbic acid, riboflavin, citric acid and sodium bisulphit upon the mitotic division of Larix decidua Mill ssp. carpatica, Picea abies (L. Karst and Thuja orientalis L. The treatment was applied in one variant, germinated seeds in ascorbic acid, riboflavin, citric acid and sodium bisulphit in 3 concentrations.

  12. Genetic variation in carbon isotope discrimination and its relationship to growth under field conditions in full-sib families of Picea mariana

    Science.gov (United States)

    Lawrence B. Flanagan; Kurt H. Johnsen

    1995-01-01

    Measurements of the stable carbon isotope composition of leaf tissue were made on Picea mariana (Mill.) B.S.P. trees from four full-sib families grown on three different field sites at the Petawawa National Forestry Institute, Ontario, Canada. The four families chosen exhibited genetic variation for growth characteristics. Genetic...

  13. Liming effects on the chemical composition of the organic surface layer of a mature Norway spruce stand (Picea abies [L.] Karst.)

    NARCIS (Netherlands)

    Rosenberg, W.; Nierop, K.G.J.; Knicker, H.; Jager, de P.A.; Kreutzer, K.; Weiá, T.

    2003-01-01

    The application of lime in a mature Norway spruce (Picea abies [L.] Karst.) forest in southern Germany induced major changes in the activity of soil organisms and root growth. Since this may influence the chemical compostion of the soil organic matter (SOM) of the organic surface layer, its

  14. Calcium addition at the Hubbard Brook Experimental Forest increases the capacity for stress tolerance and carbon capture in red spruce (Picea rubens) trees during the cold season

    Science.gov (United States)

    Paul G. Schaberg; Rakesh Minocha; Stephanie Long; Joshua M. Halman; Gary J. Hawley; Christopher. Eagar

    2011-01-01

    Red spruce (Picea rubens Sarg.) trees are uniquely vulnerable to foliar freezing injury during the cold season (fall and winter), but are also capable of photosynthetic activity if temperatures moderate. To evaluate the influence of calcium (Ca) addition on the physiology of red spruce during the cold season, we measured concentrations of foliar...

  15. Changes in phytochelatins and their biosynthetic intermediates in red spruce (Picea rubens Sarg.) cell suspension cultures under cadmium and zinc stress

    Science.gov (United States)

    P. Thangavel; Stephanie Long; Rakesh Minocha

    2007-01-01

    Cell suspension cultures of red spruce (Picea rubens Sarg.) were selected to study the effects of cadmium (Cd) and zinc (Zn) on phytochelatins (PCs) and related metabolites after 24 h exposure. The PC2 and its precursor, γ-glutamylcysteine (γ-EC) increased two to fourfold with Cd concentrations ranging from 12...

  16. Suicidal autointegration of sleeping beauty and piggyBac transposons in eukaryotic cells.

    Directory of Open Access Journals (Sweden)

    Yongming Wang

    2014-03-01

    Full Text Available Transposons are discrete segments of DNA that have the distinctive ability to move and replicate within genomes across the tree of life. 'Cut and paste' DNA transposition involves excision from a donor locus and reintegration into a new locus in the genome. We studied molecular events following the excision steps of two eukaryotic DNA transposons, Sleeping Beauty (SB and piggyBac (PB that are widely used for genome manipulation in vertebrate species. SB originates from fish and PB from insects; thus, by introducing these transposons to human cells we aimed to monitor the process of establishing a transposon-host relationship in a naïve cellular environment. Similarly to retroviruses, neither SB nor PB is capable of self-avoidance because a significant portion of the excised transposons integrated back into its own genome in a suicidal process called autointegration. Barrier-to-autointegration factor (BANF1, a cellular co-factor of certain retroviruses, inhibited transposon autointegration, and was detected in higher-order protein complexes containing the SB transposase. Increasing size sensitized transposition for autointegration, consistent with elevated vulnerability of larger transposons. Both SB and PB were affected similarly by the size of the transposon in three different assays: excision, autointegration and productive transposition. Prior to reintegration, SB is completely separated from the donor molecule and followed an unbiased autointegration pattern, not associated with local hopping. Self-disruptive autointegration occurred at similar frequency for both transposons, while aberrant, pseudo-transposition events were more frequently observed for PB.

  17. [Interest of a new instrument to assess cognition in schizophrenia: The Brief Assessment of Cognition in Schizophrenia (BACS)].

    Science.gov (United States)

    Bralet, M C; Navarre, M; Eskenazi, A M; Lucas-Ross, M; Falissard, B

    2008-12-01

    SCHIZOPHRENIA: It is therefore of great interest to create an available and easily used battery of validated tests. This would enable one to measure the different cognitive deficits and to repeat the tests, and assess evolution through longitudinal follow up of the patients. The BACS is a new instrument developed by Keefe et al. in the Department of Psychiatry and Behavioural Sciences at the University of Duke Medical Centre. It evaluates the cognitive dimensions specifically altered in schizophrenia and correlated with the evolution of the disease. This test is simple to use, requiring only paper, pencils and a stopwatch. It can be administered by different carers. The duration of the test session is approximately 35min. This battery of tests was validated on a sample of 150 patients compared with a sample of 50 controls, matched for age, parent education and ethnic groups. This aim of this study is to create a French adaptation of the BACS (translation and back translation approved by the Department of Psychiatry and Behavioural Sciences at the University of Duke Medical Centre) and then to test its easiness of administration and its sensitivity, performing correlation analysis between the French Version of the BACS (version A) and a standard battery. Its adaptation and validation in French would at first be useful for the French-speaking areas and then would add some new data for the pertinence of using the BACS. 35 French stabilized schizophrenic patients were recruited from the inpatient and outpatient facilities at the Clermont-de-L'Oise Mental Health Hospital (Picardie area, France) in Dr Boitard's Psychiatric Department (FJ 5.) Patients were required to meet DSM-IV criteria for schizophrenia or schizoaffective illness. The patients were tested on two separate days by two independent clinicians with less than two weeks between the two assessments. During the first test session, subjects received the French A version of the BACS and during the second session, they were

  18. Sequencing of a QTL-rich region of the Theobroma cacao genome using pooled BACs and the identification of trait specific candidate genes

    Directory of Open Access Journals (Sweden)

    Blackmon Barbara P

    2011-07-01

    Full Text Available Abstract Background BAC-based physical maps provide for sequencing across an entire genome or a selected sub-genomic region of biological interest. Such a region can be approached with next-generation whole-genome sequencing and assembly as if it were an independent small genome. Using the minimum tiling path as a guide, specific BAC clones representing the prioritized genomic interval are selected, pooled, and used to prepare a sequencing library. Results This pooled BAC approach was taken to sequence and assemble a QTL-rich region, of ~3 Mbp and represented by twenty-seven BACs, on linkage group 5 of the Theobroma cacao cv. Matina 1-6 genome. Using various mixtures of read coverages from paired-end and linear 454 libraries, multiple assemblies of varied quality were generated. Quality was assessed by comparing the assembly of 454 reads with a subset of ten BACs individually sequenced and assembled using Sanger reads. A mixture of reads optimal for assembly was identified. We found, furthermore, that a quality assembly suitable for serving as a reference genome template could be obtained even with a reduced depth of sequencing coverage. Annotation of the resulting assembly revealed several genes potentially responsible for three T. cacao traits: black pod disease resistance, bean shape index, and pod weight. Conclusions Our results, as with other pooled BAC sequencing reports, suggest that pooling portions of a minimum tiling path derived from a BAC-based physical map is an effective method to target sub-genomic regions for sequencing. While we focused on a single QTL region, other QTL regions of importance could be similarly sequenced allowing for biological discovery to take place before a high quality whole-genome assembly is completed.

  19. A simple, high throughput method to locate single copy sequences from Bacterial Artificial Chromosome (BAC libraries using High Resolution Melt analysis

    Directory of Open Access Journals (Sweden)

    Caligari Peter DS

    2010-05-01

    Full Text Available Abstract Background The high-throughput anchoring of genetic markers into contigs is required for many ongoing physical mapping projects. Multidimentional BAC pooling strategies for PCR-based screening of large insert libraries is a widely used alternative to high density filter hybridisation of bacterial colonies. To date, concerns over reliability have led most if not all groups engaged in high throughput physical mapping projects to favour BAC DNA isolation prior to amplification by conventional PCR. Results Here, we report the first combined use of Multiplex Tandem PCR (MT-PCR and High Resolution Melt (HRM analysis on bacterial stocks of BAC library superpools as a means of rapidly anchoring markers to BAC colonies and thereby to integrate genetic and physical maps. We exemplify the approach using a BAC library of the model plant Arabidopsis thaliana. Super pools of twenty five 384-well plates and two-dimension matrix pools of the BAC library were prepared for marker screening. The entire procedure only requires around 3 h to anchor one marker. Conclusions A pre-amplification step during MT-PCR allows high multiplexing and increases the sensitivity and reliability of subsequent HRM discrimination. This simple gel-free protocol is more reliable, faster and far less costly than conventional PCR screening. The option to screen in parallel 3 genetic markers in one MT-PCR-HRM reaction using templates from directly pooled bacterial stocks of BAC-containing bacteria further reduces time for anchoring markers in physical maps of species with large genomes.

  20. INTERACTION BETWEEN DIFFERENT MOLECULAR FORMS OF IMMUNOGLOBULIN A AND RECOMBINANT DERIVATIVES POLYPEPTIDES OF BAC RECEPTOR PROTEINS FROM GROUP B STREPTOCOCCI

    Directory of Open Access Journals (Sweden)

    A. S. Korzhueva

    2008-01-01

    Full Text Available Abstract. The article concerns interactions between immunoglobulin A and recombinant P6, P7, P8 polypeptides, designed on the basis of externally localized Bac protein of the Group B streptococci, possessing IgA-binding activity.There is a current demand for immunochemical reagents that are strictly specific for IgA, in order to develop antigenic standards for detection of IgA levels in biological fluids, as well as for affinity purification of IgA and its fragments.To analyze an opportunity of the abovementioned application ways for these proteins, a special study was performed to assay an interaction capability of recombinant P6, P7, P8 polypeptides binding to Fc regions of different IgA forms (serum IgA, secretory IgA, subclasses of serum IgA – IgA1, IgA2. Selectivity of ligand binding was specially confirmed.It was found out that, among three presented polypeptides, the structure of recombinant P6 derivative proved to be optimal for IgA-binding ability of Bac protein.Structural features of IgA-binding fragments of Bac protein, i.e., binding site position on the IgA molecule (proximity to epitopes for three monoclonal antibodies, variability of the site structure, as well as resistance of binding site for P6, P7, P8 in IgA molecule against partial disulfide bonds reduction. (Med. Immunol., vol. 10, N 4-5, pp 327-336.

  1. Toxicity and genotoxicity of the quaternary ammonium compound benzalkonium chloride (BAC) using Daphnia magna and Ceriodaphnia dubia as model systems

    International Nuclear Information System (INIS)

    Lavorgna, Margherita; Russo, Chiara; D'Abrosca, Brigida; Parrella, Alfredo; Isidori, Marina

    2016-01-01

    The toxicity and genotoxicity of the cationic surfactant benzalkonium chloride (BAC) were studied using Daphnia magna and Ceriodaphnia dubia as model systems. Acute and chronic toxicity testing were performed according to the international standard guidelines and the genotoxicity was detected through the comet assay on cells from whole organisms in vivo exposed. Acute effects occurred at concentrations in the order of tens of μg/L in D. magna and hundreds of μg/L in C. dubia. Chronic effects were found at one order of magnitude less than short-term effects maintaining the same difference in sensitivity between D. magna and C. dubia. BAC induced relevant DNA damage, in both cladocerans; the lowest adverse effect levels were 0.4 and 4 ng/L for D. magna and C. dubia, respectively. As these effective concentrations are far lower than BAC occurrence in surface waters (units of μg/L) a concerning environmental risk cannot be excluded. The findings of this study showed that D. magna and C. dubia, could be used as model organisms to detect acute and chronic toxicity as well as genotoxicity at the whole organism level. - Highlights: • Benzalkonium chloride chronic effect in C. dubia was found at dozens of μg/L. • The LOAEC detected by comet assay in D. magna is in the order of hundreds of pg/L. • D. magna and C. dubia are useful model organisms to detect toxicity and genotoxicity. - Benzalkonium chloride showed chronic toxicity and genotoxicity in Daphnia magna and Ceriodaphnia dubia at concentrations of environmental concern. Daphnids are useful model organisms.

  2. Comparative BAC-based mapping in the white-throated sparrow, a novel behavioral genomics model, using interspecies overgo hybridization

    Directory of Open Access Journals (Sweden)

    Gonser Rusty A

    2011-06-01

    Full Text Available Abstract Background The genomics era has produced an arsenal of resources from sequenced organisms allowing researchers to target species that do not have comparable mapping and sequence information. These new "non-model" organisms offer unique opportunities to examine environmental effects on genomic patterns and processes. Here we use comparative mapping as a first step in characterizing the genome organization of a novel animal model, the white-throated sparrow (Zonotrichia albicollis, which occurs as white or tan morphs that exhibit alternative behaviors and physiology. Morph is determined by the presence or absence of a complex chromosomal rearrangement. This species is an ideal model for behavioral genomics because the association between genotype and phenotype is absolute, making it possible to identify the genomic bases of phenotypic variation. Findings We initiated a genomic study in this species by characterizing the white-throated sparrow BAC library via filter hybridization with overgo probes designed for the chicken, turkey, and zebra finch. Cross-species hybridization resulted in 640 positive sparrow BACs assigned to 77 chicken loci across almost all macro-and microchromosomes, with a focus on the chromosomes associated with morph. Out of 216 overgos, 36% of the probes hybridized successfully, with an average number of 3.0 positive sparrow BACs per overgo. Conclusions These data will be utilized for determining chromosomal architecture and for fine-scale mapping of candidate genes associated with phenotypic differences. Our research confirms the utility of interspecies hybridization for developing comparative maps in other non-model organisms.

  3. Polyphenolic enriched extract of Cassia glauca Lamk, improves streptozotocin-induced type-1 diabetes linked with partial insulin resistance in rats.

    Science.gov (United States)

    Veerapur, V P; Pratap, V; Thippeswamy, B S; Marietta, P; Bansal, Punit; Kulkarni, P V; Kulkarni, V H

    2017-02-23

    Traditionally Cassia glauca (CG) has been used to treat diabetes. The study was undertaken to evaluate anti-diabetic and antioxidant activity of polyphenolic enriched extract of CG in standardized streptozotocin (STZ)-induced diabetic rats. The effect of ethanol (CGE) and water (CGW) extracts of CG (200 and 400mg/kg) treatment were evaluated in STZ (50mg/kg, iv) induced diabetic rats. On 10 th day, oral glucose tolerance test and degree of insulin resistance was calculated. On 13 th day, insulin tolerance test was performed to know the peripheral utilization of glucose. On 15 th day, blood glucose, lipid profiles and endogenous antioxidant levels were estimated. In addition, the effects on oral glucose/sucrose tolerance test in normal rats. Further, HPLC fingerprinting profile of CGE and simultaneous quantification of biomarkers were carried out. Supplementation with CGE and CGW significantly reduced STZ-induced deleterious effects and improved glucose tolerance, and insulin tolerance. In addition, supplementation also decreased oxidative stress by improving endogenous antioxidant levels. Furthermore, administration significantly improves sucrose tolerance suggesting that extract possess inhibition of α-glucosidase enzyme. Further, HPLC studies revealed that CGE contains three bioactive polyphenolic compounds viz., rutin (0.10±0.01mg/g), luteolin-7-glucoside (0.06±0.01mg/g) and isorhoifolin (0.7±0.05mg/g). Observed beneficial outcome of CG might be attributed to the presence of polyphenolic compounds and mediated by interacting with multiple targets of diabetes and oxidative stress. Taken together, this study provided the scientific evidence for the traditional use of CG. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  4. Genetic differentiation and phylogeography of Mediterranean-North Eastern Atlantic blue shark (Prionace glauca, L. 1758 using mitochondrial DNA: panmixia or complex stock structure?

    Directory of Open Access Journals (Sweden)

    Agostino Leone

    2017-12-01

    Full Text Available Background The blue shark (Prionace glauca, Linnaeus 1758 is one of the most abundant epipelagic shark inhabiting all the oceans except the poles, including the Mediterranean Sea, but its genetic structure has not been confirmed at basin and interoceanic distances. Past tagging programs in the Atlantic Ocean failed to find evidence of migration of blue sharks between the Mediterranean and the adjacent Atlantic, despite the extreme vagility of the species. Although the high rate of by-catch in the Mediterranean basin, to date no genetic study on Mediterranean blue shark was carried out, which constitutes a significant knowledge gap, considering that this population is classified as “Critically Endangered”, unlike its open-ocean counterpart. Methods Blue shark phylogeography and demography in the Mediterranean Sea and North-Eastern Atlantic Ocean were inferred using two mitochondrial genes (Cytb and control region amplified from 207 and 170 individuals respectively, collected from six localities across the Mediterranean and two from the North-Eastern Atlantic. Results Although no obvious pattern of geographical differentiation was apparent from the haplotype network, Φst analyses indicated significant genetic structure among four geographical groups. Demographic analyses suggest that these populations have experienced a constant population expansion in the last 0.4–0.1 million of years. Discussion The weak, but significant, differences in Mediterranean and adjacent North-eastern Atlantic blue sharks revealed a complex phylogeographic structure, which appears to reject the assumption of panmixia across the study area, but also supports a certain degree of population connectivity across the Strait of Gibraltar, despite the lack of evidence of migratory movements observed by tagging data. Analyses of spatial genetic structure in relation to sex-ratio and size could indicate some level of sex/stage biased migratory behaviour.

  5. New genomic resources for switchgrass: a BAC library and comparative analysis of homoeologous genomic regions harboring bioenergy traits

    Directory of Open Access Journals (Sweden)

    Feltus Frank A

    2011-07-01

    Full Text Available Abstract Background Switchgrass, a C4 species and a warm-season grass native to the prairies of North America, has been targeted for development into an herbaceous biomass fuel crop. Genetic improvement of switchgrass feedstock traits through marker-assisted breeding and biotechnology approaches calls for genomic tools development. Establishment of integrated physical and genetic maps for switchgrass will accelerate mapping of value added traits useful to breeding programs and to isolate important target genes using map based cloning. The reported polyploidy series in switchgrass ranges from diploid (2X = 18 to duodecaploid (12X = 108. Like in other large, repeat-rich plant genomes, this genomic complexity will hinder whole genome sequencing efforts. An extensive physical map providing enough information to resolve the homoeologous genomes would provide the necessary framework for accurate assembly of the switchgrass genome. Results A switchgrass BAC library constructed by partial digestion of nuclear DNA with EcoRI contains 147,456 clones covering the effective genome approximately 10 times based on a genome size of 3.2 Gigabases (~1.6 Gb effective. Restriction digestion and PFGE analysis of 234 randomly chosen BACs indicated that 95% of the clones contained inserts, ranging from 60 to 180 kb with an average of 120 kb. Comparative sequence analysis of two homoeologous genomic regions harboring orthologs of the rice OsBRI1 locus, a low-copy gene encoding a putative protein kinase and associated with biomass, revealed that orthologous clones from homoeologous chromosomes can be unambiguously distinguished from each other and correctly assembled to respective fingerprint contigs. Thus, the data obtained not only provide genomic resources for further analysis of switchgrass genome, but also improve efforts for an accurate genome sequencing strategy. Conclusions The construction of the first switchgrass BAC library and comparative analysis of

  6. Comparison of BacT/Alert FAN and FAN Plus Bottles with Conventional Medium for Culturing Cerebrospinal Fluid.

    Science.gov (United States)

    Yoo, In Young; Chun, Sejong; Song, Dong Joon; Huh, Hee Jae; Lee, Nam Yong

    2016-11-01

    We compared the BacT/Alert system FAN and FAN Plus media to conventional media for culturing cerebrospinal fluid (CSF) with 2,545 samples. FAN/FAN Plus bottles showed better performance for isolating microorganisms in CSF than conventional media (positive rate, 7.2% [182/2,545] versus 3.1% [80/2,545]). The incremental recovery rate of Cryptococcus neoformans from FAN Plus bottles was higher than that from FAN bottles. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  7. A Bac Library and Paired-PCR Approach to Mapping and Completing the Genome Sequence of Sulfolobus Solfataricus P2

    DEFF Research Database (Denmark)

    She, Qunxin; Confalonieri, F.; Zivanovic, Y.

    2000-01-01

    The original strategy used in the Sulfolobus solfatnricus genome project was to sequence non overlapping, or minimally overlapping, cosmid or lambda inserts without constructing a physical map. However, after only about two thirds of the genome sequence was completed, this approach became counter......-productive because there was a high sequence bias in the cosmid and lambda libraries. Therefore, a new approach was devised for linking the sequenced regions which may be generally applicable. BAC libraries were constructed and terminal sequences of the clones were determined and used for both end mapping and PCR...

  8. Formation and loss of large, unstable tandem arrays of the piggyBac transposable element in the yellow fever mosquito, Aedes aegypti.

    Science.gov (United States)

    Adelman, Zach N; Jasinskiene, Nijole; Vally, K J M; Peek, Corrie; Travanty, Emily A; Olson, Ken E; Brown, Susan E; Stephens, Janice L; Knudson, Dennis L; Coates, Craig J; James, Anthony A

    2004-10-01

    The Class II transposable element, piggyBac, was used to transform the yellow fever mosquito, Aedes aegypti. In two transformed lines only 15-30% of progeny inherited the transgene, with these individuals displaying mosaic expression of the EGFP marker gene. Southern analyses, gene amplification of genomic DNA, and plasmid rescue experiments provided evidence that these lines contained a high copy number of piggyBac transformation constructs and that much of this DNA consisted of both donor and helper plasmids. A detailed analysis of one line showed that the majority of piggyBac sequences were unit-length donor or helper plasmids arranged in a large tandem array that could be lost en masse in a single generation. Despite the presence of a transposase source and many intact donor elements, no conservative (cut and paste) transposition of piggyBac was observed in these lines. These results reveal one possible outcome of uncontrolled and/or unexpected recombination in this mosquito, and support the conclusion that further investigation is necessary before transposable elements such as piggyBac can be used as genetic drive mechanisms to move pathogen-resistance genes into mosquito populations.

  9. Defining “Binge” Drinking as Five Drinks per Occasion or Drinking to a 0.08% BAC: Which is More Sensitive to Risk?

    Science.gov (United States)

    Fillmore, Mark T.; Jude, Rebecca

    2011-01-01

    Heavy episodic or “binge” drinking is commonly defined as drinking 4–5 drinks per occasion (5/4 definition) or drinking that results in a blood alcohol concentration (BAC) of 0.08%. The present study compared the validity of each binge definition as an indicator of at-risk, problem drinking. 251 college students were classified as non-binge drinkers or as binge drinkers based on the 5/4 definition or the 0.08% BAC definition. The two definitions of binge drinking were examined in terms of their sensitivity and specificity as indicators of alcohol-related problems as determined by scores on the Alcohol Use Disorders Identification Test (AUDIT). Over half the sample (56%) were at-risk drinkers according to the AUDIT. The 0.08% definition detected only one-half of these individuals. Gender differences were also evident. Female binge drinkers actually achieved significantly higher estimated BACs per episode than their male binge drinking counterparts. The findings suggest that drinking to a sub-threshold BAC (i.e., risk independent of the BAC achieved during drinking episodes. The findings also highlight the importance of considering frequency of consumption in determining risky drinking versus relying solely on quantity measures. PMID:21838847

  10. Spatial attributes of the four-helix bundle group of bacteriocins – The high-resolution structure of BacSp222 in solution

    KAUST Repository

    Nowakowski, Michał

    2017-11-01

    BacSp222 is a multifunctional bacteriocin produced by Staphylococcus pseudintermedius strain 222, an opportunistic pathogen of domestic animals. At micromolar concentrations, BacSp222 kills Gram-positive bacteria and is cytotoxic toward mammalian cells, while at nanomolar doses, it acts as an immunomodulatory factor, enhancing nitric oxide release in macrophage-like cell lines. The bacteriocin is a cationic, N-terminally formylated, 50-amino-acid-long linear peptide that is rich in tryptophan residues.In this study, the solution structure of BacSp222 was determined and compared to the currently known structures of similar bacteriocins. BacSp222 was isolated from a liquid culture medium in a uniformly 13C- and 15N-labeled form, and NMR data were collected. The structure was calculated based on NMR-derived constraints and consists of a rigid and tightly packed globular bundle of four alpha-helices separated by three short turns.Although the amino acid sequence of BacSp222 has no significant similarity to any known peptide or protein, a 3D structure similarity search indicates a close relation to other four-helix bundle-motif bacteriocins, such as aureocin A53, lacticin Q and enterocins 7A/7B. Assuming similar functions, biology, structure and physicochemical properties, we propose to distinguish the four-helix bundle bacteriocins as a new Type A in subclass IId of bacteriocins, containing linear, non-pediocin-like peptides.

  11. Mobilization and integration of bacterial phenotypic data-Enabling next generation biodiversity analysis through the BacDive metadatabase.

    Science.gov (United States)

    Reimer, Lorenz C; Söhngen, Carola; Vetcininova, Anna; Overmann, Jörg

    2017-11-10

    Microbial data and metadata are scattered throughout the scientific literature, databases and unpublished lab notes and thereby often are difficult to access. Hot spots of (meta)data are internal descriptions of culture collections and initial descriptions of novel taxa in primary literature. Here we describe three exemplary mobilization projects which yielded metadata published through the prokaryotic metadatabase BacDive. The Reichenbach collection of myxobacteria includes information on 12,535 typewritten index cards which were digitized. A total of 37,156 data points were extracted by text mining. In the second mobilization project, Analytical Profile Index (API) tests on paper forms were targeted. Overall 6820 API tests were digitized, which provide physiological data of 4524 microbial strains. Thirdly, the extraction of metadata from 523 new species descriptions of the International Journal of Systematic and Evolutionary Microbiology, yielding 35,651 data points, is described. All data sets were integrated and published in BacDive. Thereby these metadata not only became accessible and searchable but were also linked to strain taxonomy, isolation source, cultivation condition, and molecular biology data. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  12. Direct conversion of human pluripotent stem cells into cranial motor neurons using a piggyBac vector

    Directory of Open Access Journals (Sweden)

    Riccardo De Santis

    2018-05-01

    Full Text Available Human pluripotent stem cells (PSCs are widely used for in vitro disease modeling. One of the challenges in the field is represented by the ability of converting human PSCs into specific disease-relevant cell types. The nervous system is composed of a wide variety of neuronal types with selective vulnerability in neurodegenerative diseases. This is particularly relevant for motor neuron diseases, in which different motor neurons populations show a different susceptibility to degeneration. Here we developed a fast and efficient method to convert human induced Pluripotent Stem Cells into cranial motor neurons of the branchiomotor and visceral motor subtype. These populations represent the motor neuron subgroup that is primarily affected by a severe form of amyotrophic lateral sclerosis with bulbar onset and worst prognosis. This goal was achieved by stable integration of an inducible vector, based on the piggyBac transposon, allowing controlled activation of Ngn2, Isl1 and Phox2a (NIP. The NIP module effectively produced electrophysiologically active cranial motor neurons. Our method can be easily extended to PSCs carrying disease-associated mutations, thus providing a useful tool to shed light on the cellular and molecular bases of selective motor neuron vulnerability in pathological conditions. Keywords: Spinal motor neuron, Cranial motor neuron, Induced pluripotent stem cells, Amyotrophic lateral sclerosis, Phox2a, piggyBac

  13. Transposon Invasion of the Paramecium Germline Genome Countered by a Domesticated PiggyBac Transposase and the NHEJ Pathway

    Science.gov (United States)

    Dubois, Emeline; Bischerour, Julien; Marmignon, Antoine; Mathy, Nathalie; Régnier, Vinciane; Bétermier, Mireille

    2012-01-01

    Sequences related to transposons constitute a large fraction of extant genomes, but insertions within coding sequences have generally not been tolerated during evolution. Thanks to their unique nuclear dimorphism and to their original mechanism of programmed DNA elimination from their somatic nucleus (macronucleus), ciliates are emerging model organisms for the study of the impact of transposable elements on genomes. The germline genome of the ciliate Paramecium, located in its micronucleus, contains thousands of short intervening sequences, the IESs, which interrupt 47% of genes. Recent data provided support to the hypothesis that an evolutionary link exists between Paramecium IESs and Tc1/mariner transposons. During development of the macronucleus, IESs are excised precisely thanks to the coordinated action of PiggyMac, a domesticated piggyBac transposase, and of the NHEJ double-strand break repair pathway. A PiggyMac homolog is also required for developmentally programmed DNA elimination in another ciliate, Tetrahymena. Here, we present an overview of the life cycle of these unicellular eukaryotes and of the developmentally programmed genome rearrangements that take place at each sexual cycle. We discuss how ancient domestication of a piggyBac transposase might have allowed Tc1/mariner elements to spread throughout the germline genome of Paramecium, without strong counterselection against insertion within genes. PMID:22888464

  14. BACs-on-Beads Technology: A Reliable Test for Rapid Detection of Aneuploidies and Microdeletions in Prenatal Diagnosis

    Directory of Open Access Journals (Sweden)

    Sandra García-Herrero

    2014-01-01

    Full Text Available The risk of fetal aneuploidies is usually estimated based on high resolution ultrasound combined with biochemical determination of criterion in maternal blood, with invasive procedures offered to the population at risk. The purpose of this study was to investigate the effectiveness of a new rapid aneuploidy screening test on amniotic fluid (AF or chorionic villus (CV samples based on BACs-on-Beads (BoBs technology and to compare the results with classical karyotyping by Giemsa banding (G-banding of cultured cells in metaphase as the gold standard technique. The prenatal-BoBs kit was used to study aneuploidies involving chromosomes 13, 18, 21, X, and Y as well as nine microdeletion syndromes in 321 AF and 43 CV samples. G-banding of metaphase cultured cells was performed concomitantly for all prenatal samples. A microarray-based comparative genomic hybridization (aCGH was also carried out in a subset of samples. Prenatal-BoBs results were widely confirmed by classical karyotyping. Only six karyotype findings were not identified by Prenatal-BoBs, all of them due to the known limitations of the technique. In summary, the BACs-on-Beads technology was an accurate, robust, and efficient method for the rapid diagnosis of common aneuploidies and microdeletion syndromes in prenatal samples.

  15. Detecting single DNA copy number variations in complex genomes using one nanogram of starting DNA and BAC-array CGH.

    Science.gov (United States)

    Guillaud-Bataille, Marine; Valent, Alexander; Soularue, Pascal; Perot, Christine; Inda, Maria Mar; Receveur, Aline; Smaïli, Sadek; Roest Crollius, Hugues; Bénard, Jean; Bernheim, Alain; Gidrol, Xavier; Danglot, Gisèle

    2004-07-29

    Comparative genomic hybridization to bacterial artificial chromosome (BAC)-arrays (array-CGH) is a highly efficient technique, allowing the simultaneous measurement of genomic DNA copy number at hundreds or thousands of loci, and the reliable detection of local one-copy-level variations. We report a genome-wide amplification method allowing the same measurement sensitivity, using 1 ng of starting genomic DNA, instead of the classical 1 microg usually necessary. Using a discrete series of DNA fragments, we defined the parameters adapted to the most faithful ligation-mediated PCR amplification and the limits of the technique. The optimized protocol allows a 3000-fold DNA amplification, retaining the quantitative characteristics of the initial genome. Validation of the amplification procedure, using DNA from 10 tumour cell lines hybridized to BAC-arrays of 1500 spots, showed almost perfectly superimposed ratios for the non-amplified and amplified DNAs. Correlation coefficients of 0.96 and 0.99 were observed for regions of low-copy-level variations and all regions, respectively (including in vivo amplified oncogenes). Finally, labelling DNA using two nucleotides bearing the same fluorophore led to a significant increase in reproducibility and to the correct detection of one-copy gain or loss in >90% of the analysed data, even for pseudotriploid tumour genomes.

  16. Enhanced Expression of Anti-CD19 Chimeric Antigen Receptor in piggyBac Transposon-Engineered T Cells

    Directory of Open Access Journals (Sweden)

    Daisuke Morita

    2018-03-01

    Full Text Available Adoptive T cell therapy using chimeric antigen receptor (CAR-modified T cells is a promising cancer immunotherapy. We previously developed a non-viral method of gene transfer into T cells using a piggyBac transposon system to improve the cost-effectiveness of CAR-T cell therapy. Here, we have further improved our technology by a novel culture strategy to increase the transfection efficiency and to reduce the time of T cell manufacturing. Using a CH2CH3-free CD19-specific CAR transposon vector and combining irradiated activated T cells (ATCs as feeder cells and virus-specific T cell receptor (TCR stimulation, we achieved 51.4% ± 14% CAR+ T cells and 2.8-fold expansion after 14 culture days. Expanded CD19.CAR-T cells maintained a significant fraction of CD45RA+CCR7+ T cells and demonstrated potent antitumor activity against CD19+ leukemic cells both in vitro and in vivo. Therefore, piggyBac-based gene transfer may provide an alternative to viral gene transfer for CAR-T cell therapy.

  17. Forestry adaptation measures at the decline of Norway spruce (Picea abies Karst. stands as exemplified by the Silesian Beskids, Czech Republic

    Directory of Open Access Journals (Sweden)

    Petr Čermák

    2011-01-01

    Full Text Available At the beginning of this century, particularly after 2003, decline of Picea abies occurred at Forest District Jablunkov in the Silesian Beskids. This decline is of the complex character disease caused by the synergetic effects of abiotic, biotic and anthropogenic factors. Under conditions of climatic changes, it is possible to expect that similar episodes will repeat and appear also in other regions. Forestry will have to respond to them by changes in forest management. Measures proposed and discussed in this paper can be a starting point in their basic principles for other similar regions. Fundamental spheres of possible measures are as follows: chemical adaptations of the soil environment, i.e. liming or fertilization (if to realise them or not, changes in the species composition (particularly the rate of the decrease of Picea abies, participation of Fagus sylvatica and increasing the diversity of tree species, modification of the rotation (decrease and regeneration period (increase.

  18. Contributions of Bacău to the economic literature and culture – The life and work of Professor Vasile Pătruţ

    OpenAIRE

    Mihai Deju

    2012-01-01

    The economic culture and accounting theory in Bacău area has its beginnings in the setting up of the first practical school of agriculture, by Ion Ionescu de la Brad, who included in the curriculum an accounting course, as well. Over the years, the economic education of Bacău area education has evolved from “The School of Accounting and Co-operative Education” (1919) to the modern economic higher education, in our days. During an important period of the evolution of education and culture in B...

  19. Third-Generation Sequencing and Analysis of Four Complete Pig Liver Esterase Gene Sequences in Clones Identified by Screening BAC Library.

    Science.gov (United States)

    Zhou, Qiongqiong; Sun, Wenjuan; Liu, Xiyan; Wang, Xiliang; Xiao, Yuncai; Bi, Dingren; Yin, Jingdong; Shi, Deshi

    2016-01-01

    Pig liver carboxylesterase (PLE) gene sequences in GenBank are incomplete, which has led to difficulties in studying the genetic structure and regulation mechanisms of gene expression of PLE family genes. The aim of this study was to obtain and analysis of complete gene sequences of PLE family by screening from a Rongchang pig BAC library and third-generation PacBio gene sequencing. After a number of existing incomplete PLE isoform gene sequences were analysed, primers were designed based on conserved regions in PLE exons, and the whole pig genome used as a template for Polymerase chain reaction (PCR) amplification. Specific primers were then selected based on the PCR amplification results. A three-step PCR screening method was used to identify PLE-positive clones by screening a Rongchang pig BAC library and PacBio third-generation sequencing was performed. BLAST comparisons and other bioinformatics methods were applied for sequence analysis. Five PLE-positive BAC clones, designated BAC-10, BAC-70, BAC-75, BAC-119 and BAC-206, were identified. Sequence analysis yielded the complete sequences of four PLE genes, PLE1, PLE-B9, PLE-C4, and PLE-G2. Complete PLE gene sequences were defined as those containing regulatory sequences, exons, and introns. It was found that, not only did the PLE exon sequences of the four genes show a high degree of homology, but also that the intron sequences were highly similar. Additionally, the regulatory region of the genes contained two 720bps reverse complement sequences that may have an important function in the regulation of PLE gene expression. This is the first report to confirm the complete sequences of four PLE genes. In addition, the study demonstrates that each PLE isoform is encoded by a single gene and that the various genes exhibit a high degree of sequence homology, suggesting that the PLE family evolved from a single ancestral gene. Obtaining the complete sequences of these PLE genes provides the necessary foundation for

  20. Linking foliar chemistry to forest floor solid and solution phase organic C and N in Picea ahies [L.) Karst stands in northern Bohemia

    Czech Academy of Sciences Publication Activity Database

    Aitkenhead-Peterson, J. A.; Alexander, J.E.; Albrechtová, J.; Krám, P.; Rock, B.; Cudlín, Pavel; Hruška, J.; Lhotáková, Z.; Huntley, R.; Oulehle, F.; Polák, T.; McDowel, W.H.

    2006-01-01

    Roč. 283, 1-2 (2006), s. 187-201 ISSN 0032-079X R&D Projects: GA MŠk ME 658 Institutional research plan: CEZ:AV0Z60870520 Keywords : cellulose and nitrogen * dissolved organic carbon * dissolved organic nitrogen * forest floor C:N * foliar Iignin, * Picea abies [L.] Karst Subject RIV: GK - Forestry Impact factor: 1.495, year: 2006

  1. Predicting Effects of Climate Change on Habitat Suitability of Red Spruce (Picea rubens Sarg.) in the Southern Appalachian Mountains of the USA: Understanding Complex Systems Mechanisms through Modeling

    OpenAIRE

    Koo, Kyung; Patten, Bernard; Madden, Marguerite

    2015-01-01

    Alpine, subalpine and boreal tree species, of low genetic diversity and adapted to low optimal temperatures, are vulnerable to the warming effects of global climate change. The accurate prediction of these species’ distributions in response to climate change is critical for effective planning and management. The goal of this research is to predict climate change effects on the distribution of red spruce (Picea rubens Sarg.) in the Great Smoky Mountains National Park (GSMNP), eastern USA. Clim...

  2. Needle age and season influence photosynthetic temperature response and total annual carbon uptake in mature Picea mariana trees

    Science.gov (United States)

    Jensen, Anna M.; Warren, Jeffrey M.; Hanson, Paul J.; Childs, Joanne; Wullschleger, Stan D.

    2015-01-01

    Background and Aims The carbon (C) balance of boreal terrestrial ecosystems is sensitive to increasing temperature, but the direction and thresholds of responses are uncertain. Annual C uptake in Picea and other evergreen boreal conifers is dependent on seasonal- and cohort-specific photosynthetic and respiratory temperature response functions, so this study examined the physiological significance of maintaining multiple foliar cohorts for Picea mariana trees within an ombrotrophic bog ecosystem in Minnesota, USA. Methods Measurements were taken on multiple cohorts of needles for photosynthetic capacity, foliar respiration (Rd) and leaf biochemistry and morphology of mature trees from April to October over 4 years. The results were applied to a simple model of canopy photosynthesis in order to simulate annual C uptake by cohort age under ambient and elevated temperature scenarios. Key Results Temperature responses of key photosynthetic parameters [i.e. light-saturated rate of CO2 assimilation (Asat), rate of Rubisco carboxylation (Vcmax) and electron transport rate (Jmax)] were dependent on season and generally less responsive in the developing current-year (Y0) needles compared with 1-year-old (Y1) or 2-year-old (Y2) foliage. Temperature optimums ranged from 18·7 to 23·7, 31·3 to 38·3 and 28·7 to 36·7 °C for Asat, Vcmax and Jmax, respectively. Foliar cohorts differed in their morphology and photosynthetic capacity, which resulted in 64 % of modelled annual stand C uptake from Y1&2 cohorts (LAI 0·67 m2 m−2) and just 36 % from Y0 cohorts (LAI 0·52 m2 m−2). Under warmer climate change scenarios, the contribution of Y0 cohorts was even less; e.g. 31 % of annual C uptake for a modelled 9 °C rise in mean summer temperatures. Results suggest that net annual C uptake by P. mariana could increase under elevated temperature, and become more dependent on older foliar cohorts. Conclusions Collectively, this study illustrates the physiological and

  3. Physical mapping and BAC-end sequence analysis provide initial insights into the flax (Linum usitatissimum L. genome

    Directory of Open Access Journals (Sweden)

    Cloutier Sylvie

    2011-05-01

    Full Text Available Abstract Background Flax (Linum usitatissimum L. is an important source of oil rich in omega-3 fatty acids, which have proven health benefits and utility as an industrial raw material. Flax seeds also contain lignans which are associated with reducing the risk of certain types of cancer. Its bast fibres have broad industrial applications. However, genomic tools needed for molecular breeding were non existent. Hence a project, Total Utilization Flax GENomics (TUFGEN was initiated. We report here the first genome-wide physical map of flax and the generation and analysis of BAC-end sequences (BES from 43,776 clones, providing initial insights into the genome. Results The physical map consists of 416 contigs spanning ~368 Mb, assembled from 32,025 fingerprints, representing roughly 54.5% to 99.4% of the estimated haploid genome (370-675 Mb. The N50 size of the contigs was estimated to be ~1,494 kb. The longest contig was ~5,562 kb comprising 437 clones. There were 96 contigs containing more than 100 clones. Approximately 54.6 Mb representing 8-14.8% of the genome was obtained from 80,337 BES. Annotation revealed that a large part of the genome consists of ribosomal DNA (~13.8%, followed by known transposable elements at 6.1%. Furthermore, ~7.4% of sequence was identified to harbour novel repeat elements. Homology searches against flax-ESTs and NCBI-ESTs suggested that ~5.6% of the transcriptome is unique to flax. A total of 4064 putative genomic SSRs were identified and are being developed as novel markers for their use in molecular breeding. Conclusion The first genome-wide physical map of flax constructed with BAC clones provides a framework for accessing target loci with economic importance for marker development and positional cloning. Analysis of the BES has provided insights into the uniqueness of the flax genome. Compared to other plant genomes, the proportion of rDNA was found to be very high whereas the proportion of known transposable

  4. Physical mapping and BAC-end sequence analysis provide initial insights into the flax (Linum usitatissimum L.) genome.

    Science.gov (United States)

    Ragupathy, Raja; Rathinavelu, Rajkumar; Cloutier, Sylvie

    2011-05-09

    Flax (Linum usitatissimum L.) is an important source of oil rich in omega-3 fatty acids, which have proven health benefits and utility as an industrial raw material. Flax seeds also contain lignans which are associated with reducing the risk of certain types of cancer. Its bast fibres have broad industrial applications. However, genomic tools needed for molecular breeding were non existent. Hence a project, Total Utilization Flax GENomics (TUFGEN) was initiated. We report here the first genome-wide physical map of flax and the generation and analysis of BAC-end sequences (BES) from 43,776 clones, providing initial insights into the genome. The physical map consists of 416 contigs spanning ~368 Mb, assembled from 32,025 fingerprints, representing roughly 54.5% to 99.4% of the estimated haploid genome (370-675 Mb). The N50 size of the contigs was estimated to be ~1,494 kb. The longest contig was ~5,562 kb comprising 437 clones. There were 96 contigs containing more than 100 clones. Approximately 54.6 Mb representing 8-14.8% of the genome was obtained from 80,337 BES. Annotation revealed that a large part of the genome consists of ribosomal DNA (~13.8%), followed by known transposable elements at 6.1%. Furthermore, ~7.4% of sequence was identified to harbour novel repeat elements. Homology searches against flax-ESTs and NCBI-ESTs suggested that ~5.6% of the transcriptome is unique to flax. A total of 4064 putative genomic SSRs were identified and are being developed as novel markers for their use in molecular breeding. The first genome-wide physical map of flax constructed with BAC clones provides a framework for accessing target loci with economic importance for marker development and positional cloning. Analysis of the BES has provided insights into the uniqueness of the flax genome. Compared to other plant genomes, the proportion of rDNA was found to be very high whereas the proportion of known transposable elements was low. The SSRs identified from BES will be

  5. BAC CGH-array identified specific small-scale genomic imbalances in diploid DMBA-induced rat mammary tumors

    International Nuclear Information System (INIS)

    Samuelson, Emma; Karlsson, Sara; Partheen, Karolina; Nilsson, Staffan; Szpirer, Claude; Behboudi, Afrouz

    2012-01-01

    Development of breast cancer is a multistage process influenced by hormonal and environmental factors as well as by genetic background. The search for genes underlying this malignancy has recently been highly productive, but the etiology behind this complex disease is still not understood. In studies using animal cancer models, heterogeneity of the genetic background and environmental factors is reduced and thus analysis and identification of genetic aberrations in tumors may become easier. To identify chromosomal regions potentially involved in the initiation and progression of mammary cancer, in the present work we subjected a subset of experimental mammary tumors to cytogenetic and molecular genetic analysis. Mammary tumors were induced with DMBA (7,12-dimethylbenz[a]anthrazene) in female rats from the susceptible SPRD-Cu3 strain and from crosses and backcrosses between this strain and the resistant WKY strain. We first produced a general overview of chromosomal aberrations in the tumors using conventional kartyotyping (G-banding) and Comparative Genome Hybridization (CGH) analyses. Particular chromosomal changes were then analyzed in more details using an in-house developed BAC (bacterial artificial chromosome) CGH-array platform. Tumors appeared to be diploid by conventional karyotyping, however several sub-microscopic chromosome gains or losses in the tumor material were identified by BAC CGH-array analysis. An oncogenetic tree analysis based on the BAC CGH-array data suggested gain of rat chromosome (RNO) band 12q11, loss of RNO5q32 or RNO6q21 as the earliest events in the development of these mammary tumors. Some of the identified changes appear to be more specific for DMBA-induced mammary tumors and some are similar to those previously reported in ACI rat model for estradiol-induced mammary tumors. The later group of changes is more interesting, since they may represent anomalies that involve genes with a critical role in mammary tumor development. Genetic

  6. Growth, allocation and tissue chemistry of Picea abies seedlings affected by nutrient supply during the second growing season.

    Science.gov (United States)

    Kaakinen, Seija; Jolkkonen, Annika; Iivonen, Sari; Vapaavuori, Elina

    2004-06-01

    One-year-old Norway spruce (Picea abies (L.) Karst.) seedlings were grown hydroponically in a growth chamber to investigate the effects of low and high nutrient availability (LN; 0.25 mM N and HN; 2.50 mM N) on growth, biomass allocation and chemical composition of needles, stem and roots during the second growing season. Climatic conditions in the growth chamber simulated the mean growing season from May to early October in Flakaliden, northern Sweden. In the latter half of the growing season, biomass allocation changed in response to nutrient availability: increased root growth and decreased shoot growth led to higher root/shoot ratios in LN seedlings than in HN seedlings. At high nutrient availability, total biomass, especially stem biomass, increased, as did total nonstructural carbohydrate and nitrogen contents per seedling. Responses of stem chemistry to nutrient addition differed from those of adult trees of the same provenance. In HN seedlings, concentrations of alpha-cellulose, hemicellulose and lignin decreased in the secondary xylem. Our results illustrate the significance of retranslocation of stored nutrients to support new growth early in the season when root growth and nutrient uptake are still low. We conclude that nutrient availability alters allocation patterns, thereby influencing the success of 2-year-old Norway spruce seedlings at forest planting sites.

  7. [Responses of Picea likiangensis radial growth to climate change in the Small Zhongdian area of Yunnan Province, Southwest China].

    Science.gov (United States)

    Zhao, Zhi-Jiang; Tan, Liu-Yi; Kang, Dong-Wei; Liu, Qi-Jing; Li, Jun-Qing

    2012-03-01

    Picea likiangensis (Franch. ) Pritz. primary forest is one of the dominant forest types in the Small Zhongdian area in Shangri-La County of Yunnan Province. In this paper, the responses of P. likiangensis tree-ring width to climate change were analyzed by dendrochronological methods, and the dendrochronology was built by using relatively conservative detrending negative exponential curves or linear regression. Correlation analysis and response function analysis were applied to explore the relationships between the residual chronology series (RES) and climatic factors at different time scales, and pointer year analysis was used to explain the reasons of producing narrow and wide rings. In the study area, the radial growth of P. likiangensis and the increasing air temperature from 1990 to 2008 had definite 'abruption'. The temperature and precipitation in previous year growth season were the main factors limiting the present year radial growth, and especially, the temperature in previous July played a negative feedback role in the radial growth, while the sufficient precipitation in previous July promoted the radial growth. The differences in the temperature variation and precipitation variation in previous year were the main reasons for the formation of narrow and wide rings. P. likiangensis radial growth was not sensitive to the variation of PDSI.

  8. Impact of soil drainage to the radial stem growth of Norway spruce (Picea Abies L. Karst. in peatland forests

    Directory of Open Access Journals (Sweden)

    Klempířová Barbora

    2013-12-01

    Full Text Available Peatland Norway spruce (Picea abies L. Karst. forests represent very valuable ecosystems with considerable importance for nature conservation. However, a lot of peatland forests have been drained or used for opencast mining of peat. Since dendrochronological and dendroecological studies on trees growing on peatlands in Europe are not many, this study aimed to reconstruct the impact of drainage to the growth of trees in forest stands older than 100 years in the moment of drainage. Dendrochronological analysis was performed on two 0.25-ha square sampling plots (50*50 m in two pre-selected stands (control site vs. drained site with similar natural conditions and age. The mean-value functions of the ring indices, comparing the drained site with the control site, in the period after 1940 revealed very similar radial-growth trends. After the year 1992, when one site was substantially drained, the radial-growth trends not showed any significant change. Likewise, the result of the independent two sample t-test for the period after 1992 has not revealed any substantial statistically important difference in the mean index between the control site and the drained site.

  9. Kinetics, isotherms, and thermodynamic studies of lead, chromium, and cadmium bio-adsorption from aqueous solution onto Picea smithiana sawdust.

    Science.gov (United States)

    Mahmood-Ul-Hassan, Muhammad; Yasin, Muhammad; Yousra, Munazza; Ahmad, Rizwan; Sarwar, Sair

    2018-05-01

    Lead (Pb), chromium (Cr), and cadmium (Cd) removal capacity of sawdust (Picea smithiana) from aqueous solution was investigated by conducting batch experiments. Thermodynamic parameters, like change in standard free energy (ΔG Θ ), enthalpy (ΔH Θ ) and entropy (ΔS Θ ) during bio-adsorption process were estimated using the Van't Hoff equation. The maximum metals adsorption was observed at pH 8, 20 g L -1 bio-adsorbent and at 60 min of contact time. The metal adsorption kinetics was examined by fitting the pseudo-first-order as well as four forms of pseudo-second-order kinetic models. Type 1 pseudo-second-order equation described adsorption kinetics better than others. Langmuir model and Freundlich equations were used for calculation of sorption parameters. The Langmuir maximum adsorption capacity of Pb, Cr, and Cd was 6.35, 3.37, and 2.87 mg g -1 at room temperature, respectively. The values of the separation factor (RL) were in between 0 and 1, indicating that bio-adsorption was favorable. Thermodynamics study revealed that the Pb, Cr, and Cd uptake reactions were endothermic and spontaneous. Results of the study asserted that the removal of heavy metal ions from aqueous solution is viable and the sawdust could be used in the treatment of effluents from industries, thereby reducing the level of water pollution.

  10. Wood density variations of Norway spruce (Picea abies (L. Karst. under contrasting climate conditions in southwestern Germany

    Directory of Open Access Journals (Sweden)

    Marieke van der Maaten-Theunissen

    2013-05-01

    Full Text Available We analyzed inter-annual variations in ring width and maximumwood density of Norway spruce (Picea abies (L. Karst. at different altitudes in Baden-Württemberg, southwestern Germany, to determine the climate response of these parameters under contrasting climate conditions. In addition, we compared maximum, average and minimum wood density between sites. Bootstrapped correlation coefficients of ring width and maximum wood density with monthly temperature and precipitation, revealed a different climate sensitivity of both parameters. Ring width showed strong correlations with climate variables in the previous year and in the first half of the growingseason. Further, a negative relationship with summer temperature was observed at the low-altitude sites. Maximum wood density correlated best with temperature during the growing season, whereby strongest correlations were found between September temperature and maximum wood density at the high-altitude sites. Observed differences in maximum, average and minimum wood density are suggested to relate to the local climate; with lower temperature and higher water availability having a negative effect on wood density.

  11. Wood density variations of Norway spruce (Picea abies (L. Karst. under contrasting climate conditions in southwestern Germany

    Directory of Open Access Journals (Sweden)

    Marieke van der Maaten-Theunissen

    2013-07-01

    Full Text Available We analyzed inter-annual variations in ring width and maximum wood density of Norway spruce (Picea abies (L. Karst. at different altitudes in Baden-Württemberg, southwestern Germany, to determine the climate response of these parameters under contrasting climate conditions. In addition, we compared maximum, average and minimum wood density between sites. Bootstrapped correlation coefficients of ring width and maximum wood density with monthly temperature and precipitation, revealed a different climate sensitivity of both parameters. Ring width showed strong correlations with climate variables in the previous year and in the first half of the growing season. Further, a negative relationship with summer temperature was observed at the low-altitude sites. Maximum wood density correlated best with temperature during the growing season, whereby strongest correlations were found between September temperature and maximum wood density at the high-altitude sites. Observed differences in maximum, average and minimum wood density are suggested to relate to the local climate; with lower temperatures and higher water availability having a negative effect on wood density. 

  12. Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, and growth of young spruce trees (Picea abies)

    International Nuclear Information System (INIS)

    Thomas, V.F.D.; Braun, S.; Flueckiger, W.

    2005-01-01

    Spruce saplings were grown under different nitrogen fertilization regimes in eight chamberless fumigation systems, which were fumigated with either charcoal-filtered (F) or ambient air (O 3 ). After the third growing season trees were harvested for biomass and non-structural carbohydrate analysis. Nitrogen had an overall positive effect on the investigated plant parameters, resulting in increased shoot elongation, biomass production, fine root soluble carbohydrate concentrations, and also slightly increased starch concentrations of stems and roots. Only needle starch concentrations and fine root sugar alcohol concentrations were decreased. Ozone fumigation resulted in needle discolorations and affected most parameters negatively, including decreased shoot elongation and decreased starch concentrations in roots, stems, and needles. In fine roots, however, soluble carbohydrate concentrations remained unaffected or increased by ozone fumigation. The only significant interaction was an antagonistic effect on root starch concentrations, where higher nitrogen levels alleviated the negative impact of ozone. - Simultaneous ozone fumigation and nitrogen fertilization have no synergistic impacts on carbohydrate concentrations, biomass, or growth of Picea abies saplings

  13. Static and dynamic bending has minor effects on xylem hydraulics of conifer branches (Picea abies, Pinus sylvestris).

    Science.gov (United States)

    Mayr, Stefan; Bertel, Clara; Dämon, Birgit; Beikircher, Barbara

    2014-09-01

    The xylem hydraulic efficiency and safety is usually measured on mechanically unstressed samples, although trees may be exposed to combined hydraulic and mechanical stress in the field. We analysed changes in hydraulic conductivity and vulnerability to drought-induced embolism during static bending of Picea abies and Pinus sylvestris branches as well as the effect of dynamic bending on the vulnerability. We hypothesized this mechanical stress to substantially impair xylem hydraulics. Intense static bending caused an only small decrease in hydraulic conductance (-19.5 ± 2.4% in P. abies) but no shift in vulnerability thresholds. Dynamic bending caused a 0.4 and 0.8 MPa decrease of the water potential at 50 and 88% loss of conductivity in P. sylvestris, but did not affect vulnerability thresholds in P. abies. With respect to applied extreme bending radii, effects on plant hydraulics were surprisingly small and are thus probably of minor eco-physiological importance. More importantly, results indicate that available xylem hydraulic analyses (of conifers) sufficiently reflect plant hydraulics under field conditions. © 2014 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.

  14. Modeling the Potential Distribution of Picea chihuahuana Martínez, an Endangered Species at the Sierra Madre Occidental, Mexico

    Directory of Open Access Journals (Sweden)

    Victor Aguilar-Soto

    2015-03-01

    Full Text Available Species distribution models (SDMs help identify areas for the development of populations or communities to prevent extinctions, especially in the face of the global environmental change. This study modeled the potential distribution of the tree Picea chihuahuana Martínez, a species in danger of extinction, using the maximum entropy modeling method (MaxEnt at three scales: local, state and national. We used a total of 38 presence data from the Sierra Madre Occidental. At the local scale, we compared MaxEnt with the reclassification and overlay method integrated in a geographic information system. MaxEnt generated maps with a high predictive capability (AUC > 0.97. The distribution of P. chihuahuana is defined by vegetation type and minimum temperature at national and state scales. At the local scale, both models calculated similar areas for the potential distribution of the species; the variables that better defined the species distribution were vegetation type, aspect and distance to water flows. Populations of P. chihuahuana have always been small, but our results show potential habitat greater than the area of the actual distribution. These results provide an insight into the availability of areas suitable for the species’ regeneration, possibly through assisted colonization.

  15. Normalizing gene expression by quantitative PCR during somatic embryogenesis in two representative conifer species: Pinus pinaster and Picea abies.

    Science.gov (United States)

    de Vega-Bartol, José J; Santos, Raquen Raissa; Simões, Marta; Miguel, Célia M

    2013-05-01

    Suitable internal control genes to normalize qPCR data from different stages of embryo development and germination were identified in two representative conifer species. Clonal propagation by somatic embryogenesis has a great application potentiality in conifers. Quantitative PCR (qPCR) is widely used for gene expression analysis during somatic embryogenesis and embryo germination. No single reference gene is universal, so a systematic characterization of endogenous genes for concrete conditions is fundamental for accuracy. We identified suitable internal control genes to normalize qPCR data obtained at different steps of somatic embryogenesis (embryonal mass proliferation, embryo maturation and germination) in two representative conifer species, Pinus pinaster and Picea abies. Candidate genes included endogenous genes commonly used in conifers, genes previously tested in model plants, and genes with a lower variation of the expression along embryo development according to genome-wide transcript profiling studies. Three different algorithms were used to evaluate expression stability. The geometric average of the expression values of elongation factor-1α, α-tubulin and histone 3 in P. pinaster, and elongation factor-1α, α-tubulin, adenosine kinase and CAC in P. abies were adequate for expression studies throughout somatic embryogenesis. However, improved accuracy was achieved when using other gene combinations in experiments with samples at a single developmental stage. The importance of studies selecting reference genes to use in different tissues or developmental stages within one or close species, and the instability of commonly used reference genes, is highlighted.

  16. Building Resilience into Sitka Spruce (Picea sitchensis (Bong. Carr. Forests in Scotland in Response to the Threat of Climate Change

    Directory of Open Access Journals (Sweden)

    Andrew D. Cameron

    2015-02-01

    Full Text Available It is expected that a warming climate will have an impact on the future productivity of European spruce forests. In Scotland, Sitka spruce (Picea sitchensis (Bong. Carr. dominates the commercial forestry sector and there is growing pressure to develop alternative management strategies to limit potential economic losses through climate change. This review considers management options to increase the resilience of Sitka spruce dominated forests in Scotland. Given the considerable uncertainty over the potential long-term impacts of climate change, it is recommended that Sitka spruce should continue to be planted where it already grows well. However, new planting and restocking should be established in mixtures where silviculturally practicable, even if no-thin regimes are adopted, to spread future risks of damage. Three potentially compatible species with Sitka spruce are western hemlock (Tsuga heterophylla (Raf. Sarg., grand fir (Abies grandis (Lamb. Lindl. and Douglas fir (Pseudotsuga menziesii (Mirb. Franco and all form natural mixtures in its native range in North America. The predicted windier climate will require a range of management inputs, such as early cutting of extraction racks and early selective thinning, to improve stability. The potential to improve resilience to particularly abiotic damage through transforming even-aged stands into irregular structures and limiting the overall size of the growing stock is discussed.

  17. Laboratory and field studies of biogenic volatile organic compound emissions from Sitka spruce (Picea sitchensis Bong.) in the United Kingdom

    Science.gov (United States)

    Street, Rachel A.; Duckham, S. Craig; Hewitt, C. Nicholas

    1996-10-01

    Isoprene and monoterpene emission rates were measured from Sitka spruce (Picea sitchensis Bong.) with a dynamic flow-through branch enclosure, both in the laboratory and in the field in the United Kingdom. In the laboratory, emission rates of isoprene comprised over 94% of the identified VOC species, and were exponentially related to temperature over a period of 1 day. This exponential relationship broke down at ˜33°C. Field measurements were taken on five sampling days in 1992 and 1993, in Grizedale Forest, Cumbria. Total emission rates were in the range 36-3771 ng g-1 h-1. Relative emissions were more variable than suggested by laboratory measurements, with monoterpenes contributing at least 64% to the total emissions in most cases. There was a significant variation in the basal emission rate both across the growing season and between different ages of vegetation, the causes of which are as yet unknown. Total emission rates, in July 1993, were estimated to be between 0.01 and 0.27% of assimilated carbon.

  18. Comportamiento en flexión estática de vigas de madera antigua de Picea abies

    Directory of Open Access Journals (Sweden)

    Javier Ramón Sotomayor-Castellanos

    2014-01-01

    Full Text Available Las tipologías arquitecturales encontradas en el contexto del análisis estructural de construcciones con significado histórico y cultural son complejas. El modelado numérico es propuesto como una solución para diagnosticar la seguridad estructural de estas edificaciones. En esta investigación se realizaron pruebas de flexión estática de acuerdo a la norma Europea EN 408 y se calcularon los módulos de elasticidad y de ruptura en 5 vigas antiguas de Picea abies. La densidad de la madera fue de 422 kg/m3 para las vigas con un contenido de humedad de 12 %. El módulo de elasticidad promedio fue de 9,059 MPa y el módulo de ruptura fue de 29.18 Pa. Los módulos presentaron una variabilidad aceptable. Los resultados experimentales se contrastaron con las predicciones numéricas empleando el método del elemento finito. Las predicciones numéricas en los dominios lineal y no lineal mostraron buena aproximación con los resultados experimentales.

  19. Spatial attributes of the four-helix bundle group of bacteriocins – The high-resolution structure of BacSp222 in solution

    KAUST Repository

    Nowakowski, Michał; Jaremko, Łukasz; Wladyka, Benedykt; Dubin, Grzegorz; Ejchart, Andrzej; Mak, Paweł

    2017-01-01

    collected. The structure was calculated based on NMR-derived constraints and consists of a rigid and tightly packed globular bundle of four alpha-helices separated by three short turns.Although the amino acid sequence of BacSp222 has no significant

  20. Restoring San Xavier del Bac, "Our Church": Tohono O'odham Work to Restore the 200-Year-Old Church Built by Their Ancestors.

    Science.gov (United States)

    Fontana, Bernard L.

    1995-01-01

    The Tohono O'odham built Mission San Xavier del Bac for Franciscan missionaries in the late 1700s and have protected and cared for it through changing circumstances ever since. As part of a massive restoration project, outstanding experts have been restoring the church's painted and sculpted interior and training local Tohono O'odham to be…

  1. Physical Analysis of the Complex Rye (Secale cereale L.) Alt4 Aluminium (Aluminum) Tolerance Locus Using a Whole-Genome BAC Library of Rye cv. Blanco

    Science.gov (United States)

    Rye is a diploid crop species with many outstanding qualities, and is also important as a source of new traits for wheat and triticale improvement. Here we describe a BAC library of rye cv. Blanco, representing a valuable resource for rye molecular genetic studies. The library provides a 6 × genome ...

  2. Rhipicephalus (Boophilus) microplus strain Deutsch, 5 BAC clone sequencing, including two encoding Cytochrome P450s and one encoding CzEst9 carboxylesterase

    Science.gov (United States)

    The cattle tick, Rhipicephalus (Boophilus) microplus, has a genome over 2.4 times the size of the human genome, and with over 70% of repetitive DNA, this genome would prove very costly to sequence at today's prices and difficult to assemble and analyze. BAC clones give insight into the genome struct...

  3. Inducible targeting of CNS astrocytes in Aldh1l1-CreERT2 BAC transgenic mice.

    Science.gov (United States)

    Winchenbach, Jan; Düking, Tim; Berghoff, Stefan A; Stumpf, Sina K; Hülsmann, Swen; Nave, Klaus-Armin; Saher, Gesine

    2016-01-01

    Background: Studying astrocytes in higher brain functions has been hampered by the lack of genetic tools for the efficient expression of inducible Cre recombinase throughout the CNS, including the neocortex. Methods: Therefore, we generated BAC transgenic mice, in which CreERT2 is expressed under control of the Aldh1l1 regulatory region. Results: When crossbred to Cre reporter mice, adult Aldh1l1-CreERT2 mice show efficient gene targeting in astrocytes. No such Cre-mediated recombination was detectable in CNS neurons, oligodendrocytes, and microglia. As expected, Aldh1l1-CreERT2 expression was evident in several peripheral organs, including liver and kidney. Conclusions: Taken together, Aldh1l1-CreERT2 mice are a useful tool for studying astrocytes in neurovascular coupling, brain metabolism, synaptic plasticity and other aspects of neuron-glia interactions.

  4. The IRIDICA BAC BSI Assay: Rapid, Sensitive and Culture-Independent Identification of Bacteria and Candida in Blood

    Science.gov (United States)

    Rothman, Richard E.; Peterson, Stephen; Carroll, Karen C.; Zhang, Sean X.; Avornu, Gideon D.; Rounds, Megan A.; Carolan, Heather E.; Toleno, Donna M.; Moore, David; Hall, Thomas A.; Massire, Christian; Richmond, Gregory S.; Gutierrez, Jose R.; Sampath, Rangarajan; Ecker, David J.; Blyn, Lawrence B.

    2016-01-01

    Bloodstream infection (BSI) and sepsis are rising in incidence throughout the developed world. The spread of multi-drug resistant organisms presents increasing challenges to treatment. Surviving BSI is dependent on rapid and accurate identification of causal organisms, and timely application of appropriate antibiotics. Current culture-based methods used to detect and identify agents of BSI are often too slow to impact early therapy and may fail to detect relevant organisms in many positive cases. Existing methods for direct molecular detection of microbial DNA in blood are limited in either sensitivity (likely the result of small sample volumes) or in breadth of coverage, often because the PCR primers and probes used target only a few specific pathogens. There is a clear unmet need for a sensitive molecular assay capable of identifying the diverse bacteria and yeast associated with BSI directly from uncultured whole blood samples. We have developed a method of extracting DNA from larger volumes of whole blood (5 ml per sample), amplifying multiple widely conserved bacterial and fungal genes using a mismatch- and background-tolerant PCR chemistry, and identifying hundreds of diverse organisms from the amplified fragments on the basis of species-specific genetic signatures using electrospray ionization mass spectrometry (PCR/ESI-MS). We describe the analytical characteristics of the IRIDICA BAC BSI Assay and compare its pre-clinical performance to current standard-of-care methods in a collection of prospectively collected blood specimens from patients with symptoms of sepsis. The assay generated matching results in 80% of culture-positive cases (86% when common contaminants were excluded from the analysis), and twice the total number of positive detections. The described method is capable of providing organism identifications directly from uncultured blood in less than 8 hours. Disclaimer: The IRIDICA BAC BSI Assay is not available in the United States. PMID:27384540

  5. Exploiting BAC-end sequences for the mining, characterization and utility of new short sequences repeat (SSR) markers in Citrus.

    Science.gov (United States)

    Biswas, Manosh Kumar; Chai, Lijun; Mayer, Christoph; Xu, Qiang; Guo, Wenwu; Deng, Xiuxin

    2012-05-01

    The aim of this study was to develop a large set of microsatellite markers based on publicly available BAC-end sequences (BESs), and to evaluate their transferability, discriminating capacity of genotypes and mapping ability in Citrus. A set of 1,281 simple sequence repeat (SSR) markers were developed from the 46,339 Citrus clementina BAC-end sequences (BES), of them 20.67% contained SSR longer than 20 bp, corresponding to roughly one perfect SSR per 2.04 kb. The most abundant motifs were di-nucleotide (16.82%) repeats. Among all repeat motifs (TA/AT)n is the most abundant (8.38%), followed by (AG/CT)n (4.51%). Most of the BES-SSR are located in the non-coding region, but 1.3% of BES-SSRs were found to be associated with transposable element (TE). A total of 400 novel SSR primer pairs were synthesized and their transferability and polymorphism tested on a set of 16 Citrus and Citrus relative's species. Among these 333 (83.25%) were successfully amplified and 260 (65.00%) showed cross-species transferability with Poncirus trifoliata and Fortunella sp. These cross-species transferable markers could be useful for cultivar identification, for genomic study of Citrus, Poncirus and Fortunella sp. Utility of the developed SSR marker was demonstrated by identifying a set of 118 markers each for construction of linkage map of Citrus reticulata and Poncirus trifoliata. Genetic diversity and phylogenetic relationship among 40 Citrus and its related species were conducted with the aid of 25 randomly selected SSR primer pairs and results revealed that citrus genomic SSRs are superior to genic SSR for genetic diversity and germplasm characterization of Citrus spp.

  6. [Climate-growth relationships of Picea koraiensis and causes of its recent decline in Xiaoxing' an Mountains, China].

    Science.gov (United States)

    Yao, Qi-chao; Wang, Xiao-chun; Xiao, Xing-wei

    2015-07-01

    Two tree-ring width chronologies of Picea koraiensis at two altitudes in Fenglin National Nature Reserve of Xiaoxing'an Mountains, China were developed by using dendrochronological methods. To identify main limiting factors of P. koraiensis radial growth at the two altitudes, the relationships between the chronologies and local temperature, precipitation, Palmer drought severity index (PDSI), and large-scale climatic factors were investigated. Meanwhile, the reasons of P. koraiensis growth decline in recent years were also explored. Results showed that radial growth of P. koraiensis in Xiaoxing'an Mountains was mainly limited by temperatures, especially by the minimum temperature in growing season, while the limiting effect of precipitation was relatively weak. Climate responses of P. koraiensis growth at the different altitudes showed significant differences. Radial growths of P. koraiensis at the low altitude were positively correlated with precipitation in the current growth season (June-September) and whole year, and negatively correlated with soil temperatures at different depths, especially at 80 cm depth in growing season. Meanwhile, it was signi-ficantly positively correlated with PDSI in growing season. However, the relationships between radial growth of P. koraiensis at the high altitude and precipitation, air and soil temperatures, and PDSI were not significant as that at the low altitude. Growth decline of P. koraiensis in Xiaoxing'an Mountains could be related to the phase changes in Atlantic multidecadal oscillation (AMO) and Pacific decadal oscillation (PDO) and the significant global warming since 1980. The coupling effects of the above changes might result in increased soil evaporation and exacerbated warming and drying phenomena, consequently causing the growth decline of P. koraiensis at the low altitude.

  7. Water deficit affects mesophyll limitation of leaves more strongly in sun than in shade in two contrasting Picea asperata populations.

    Science.gov (United States)

    Duan, Baoli; Li, Yan; Zhang, Xiaolu; Korpelainen, Helena; Li, Chunyang

    2009-12-01

    The aim of this study was to examine the response of internal conductance to CO(2) (g(i)) to soil water deficit and contrasting light conditions, and their consequences on photosynthetic physiology in two Picea asperata Mast. populations originating from wet and dry climate regions of China. Four-year-old trees were subjected to two light treatments (30% and 100% of full sunlight) and two watering regimes (well watered, drought) for 2 years. In both tested populations, drought significantly decreased g(i) and the net photosynthesis rate (A) and increased carbon isotope composition (delta(13)C) values in both light treatments, in particular in the sun. Moreover, drought resulted in a significantly higher relative limitation due to stomatal conductance (L(s)) in both light treatments and higher relative limitation due to internal conductance (L(i)) and abscisic acid (ABA) in the sun plants. The results also showed that L(i) (0.26-0.47) was always greater than L(s) (0.12-0.28). On the other hand, drought significantly decreased the ratio of chloroplastic to internal CO(2) concentration (C(c)/C(i)), photosynthetic nitrogen utilization efficiency (PNUE) and total biomass in the sun plants of the wet climate population, whereas there were no significant changes in these parameters in the dry climate population. Our results also showed that the dry climate population possessed higher delta(13)C values with higher ratio of internal conductance to stomatal conductance (g(i)/g(s)), suggesting that increasing the g(i)/g(s) ratio enhances water-use efficiency (WUE) in plants evolved in arid environments. Thus, we propose that the use of the g(i)/g(s) parameter to screen P. asperata plants with higher water deficit tolerance is certainly worthy of consideration. Furthermore, g(i) is an important variable, which reflects the population differences in PNUE, and it should thus be included in plant physiological investigations related to leaf economics.

  8. Variation in spread of Heterobasidion annosum in clones of Picea abies grown at different vegetation phases under greenhouse conditions

    Energy Technology Data Exchange (ETDEWEB)

    Svedjemark, G.; Stenlid, J. [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Forest Mycology and Pathology

    1996-06-01

    Forty-nine Picea abies (L.) Karst clones were inoculated under greenhouse conditions with a Heterobasidion annosum (Fr.) Bref. isolate of the S intersterility group. The cuttings were inoculated at the following vegetation stages; bud-flushing stage, vegetative stage and after bud-set. Fungal growth in sapwood and leison length in the inner bark were measured after 34 days. The susceptibility of the various clones to H. annosum was strongly correlated among the three vegetation stages, both in terms of mean growth and mean growth ranking. Partitioning of variance components showed that variation in growth was explained by physiological stages and clone to 4% and 24%, respectively, and for interaction between clone and physiological stage to 9%. Corresponding values for leison length in the inner bark were 3%, 14% and 5%, respectively. Fungal growth in wood and leison length in the inner bark were strongly correlated (r{sup 2} ranging between 0.23 and 0.36). When cuttings were inoculated during bud-flushing, leison length and fungal growth in wood were both strongly correlated with bud-flushing index of the cuttings (r{sup 2} = 0.03 and 0.04 respectively) but that was not the case for the other stages. The number of active fine-roots and the degree of wilting of the cuttings were negatively correlated with leison length and fungal growth (r{sup 2} ranging between 0.01 and 0.13). Height and diameter varied greatly between the clones and both were negatively correlated with fungal extension (r{sup 2} ranging between 0.01 and 0.09). 33 refs, 2 figs, 3 tabs

  9. Total Stem and Merchantable Volume Equations of Norway Spruce (Picea abies (L. Karst. Growing on Former Farmland in Sweden

    Directory of Open Access Journals (Sweden)

    Tord Johansson

    2014-08-01

    Full Text Available An equation was constructed to estimate the stem volume of Norway spruce (Picea abies (L. Karst. in 145 stands growing on former farmland in Sweden (Latitude 56–63° N. The mean total age was 40 ± 13 (range 17–91 years, the mean diameter at breast height (ob was 15 ± 4 (range 5–27 cm and the mean density was 1621 ± 902 (range 100–7600 stems ha−1. The equation which fits the data best used the diameter at breast height and total stem height as predictive variables. Merchantable volume equations for the estimation of commercial volume for any top diameter and bole length were developed. Soil types in the stands were sediments (coarse sand, fine sand and silt and heavy, medium and light clay, tills (sandy, fine sandy and silty and peat. The standing volume was calculated; the mean was 253 ± 103 (range 26–507 m3 ha−1 with a MAI (mean annual increment of 6.9±3.5 (range 1.3–16.7 m3 ha−1 year−1. There were statistically significant differences between MAI and coarse sand, sand and silt, light clay, peat and silty till soils. Spruce stands growing on silty tills had the lowest MAI (4.94 ± 2.27 m3 ha−1 year−1 and light clay, fine sand and silt and peat the highest (7.62 ± 4.24, 7.46 ± 3.33 and 8.67 ± 2.83 m3 ha−1 year−1.

  10. Influence of ambient air toxics in open-top chambers on the monoterpene emission of Picea abies. Diurnal and seasonal variation of emissions, and differentiation of needles and bark as emission sources. Der Einfluss natuerlich-phytotoxischer Luft auf die Monoterpen-Emission bei Picea abies in Open-Top-Kammern. Tages- und Jahresgang der Emission und Differenzierung von Nadel- und Rindenemissionen

    Energy Technology Data Exchange (ETDEWEB)

    Juettner, F. (Max-Planck-Institut fuer Limnologie, Ploen (Germany, F.R.). Abt. Oekophysiologie)

    1990-04-01

    Open-top chambers, in which each a 19-years old spruce tree (Picea abies) was growing, were used to determine monoterpene emissions by mass fragmentography. The annual dynamics of the monoterpene emissions corresponded to the air temperature. However, the diurnal dynamics did not follow the course of the temperature. Physiological reactions of the needles are responsible for the temperature independent emission of monoterpenes during the day. (orig.).

  11. Feasibility of a novel positive feedback effect of 131I-promoted Bac-Egr1-hNIS expression in malignant glioma via baculovirus

    International Nuclear Information System (INIS)

    Guo Rui; Tian Lipeng; Han Bing; Xu Haoping; Zhang Miao; Li Biao

    2011-01-01

    Purpose: As intracellular iodine is released rapidly, increased expression of sodium/iodide symporter (NIS) is required for effective radioiodine treatment of tumor. As Egr1 promoter is activated by 131 I and may promote human NIS (hNIS) expression, hNIS also induces 131 I uptake and activates Egr1, so the existence of a positive feedback effect of 131 I-promoted Egr1-hNIS expression is possible. Our purpose was to investigate the possible existence of this positive feedback effect through a series of in vitro pioneer studies. Method: Recombinant baculovirus (Bac-Egr1-hNIS) encoding the hNIS gene under the control of a radiation-inducible Egrl promoter was constructed. To test 131 I-promoted hNIS expression, human malignant glioma U87 cells were transfected with Bac-Egr1-hNIS, stimulated with or without 131 I; the expression of hNIS protein was detected by immunofluorescence and flow cytometry test. In addition, the uptake and efflux of 131 I were determined after the incubation of Bac-Egr1-hNIS-transfected U87 cells with or without 131 I. Results: Immunocytochemical staining and flow cytometry test showed a higher hNIS protein expression in Bac-Egr1-hNIS-transfected U87 cells with 131 I stimulation than in cells without stimulation. Bac-Egr1-hNIS-transfected U87 cells accumulated up to about 4.05 times of 131 I after 131 I stimulation. The amount of 131 I uptake in both groups showed a baculovirus dose-dependent manner. However, rapid efflux of radioactivity was observed in both groups, with 50% lost during the first 2 min after the 131 I-containing medium had been replaced by a nonradioactive medium. Conclusion: Our results indicated that an improved transgene expression of 131 I-stimulated hNIS in U87 cells using a baculovirus vector containing the Egr1 promoter is possible, and the increased expression of hNIS is responsible for a higher 131 I uptake. It might provide a reference for the existence of a positive feedback effect in 131 I-promoted Bac-Egr1-h

  12. Re-annotation of the physical map of Glycine max for polyploid-like regions by BAC end sequence driven whole genome shotgun read assembly

    Directory of Open Access Journals (Sweden)

    Shultz Jeffry

    2008-07-01

    Full Text Available Abstract Background Many of the world's most important food crops have either polyploid genomes or homeologous regions derived from segmental shuffling following polyploid formation. The soybean (Glycine max genome has been shown to be composed of approximately four thousand short interspersed homeologous regions with 1, 2 or 4 copies per haploid genome by RFLP analysis, microsatellite anchors to BACs and by contigs formed from BAC fingerprints. Despite these similar regions,, the genome has been sequenced by whole genome shotgun sequence (WGS. Here the aim was to use BAC end sequences (BES derived from three minimum tile paths (MTP to examine the extent and homogeneity of polyploid-like regions within contigs and the extent of correlation between the polyploid-like regions inferred from fingerprinting and the polyploid-like sequences inferred from WGS matches. Results Results show that when sequence divergence was 1–10%, the copy number of homeologous regions could be identified from sequence variation in WGS reads overlapping BES. Homeolog sequence variants (HSVs were single nucleotide polymorphisms (SNPs; 89% and single nucleotide indels (SNIs 10%. Larger indels were rare but present (1%. Simulations that had predicted fingerprints of homeologous regions could be separated when divergence exceeded 2% were shown to be false. We show that a 5–10% sequence divergence is necessary to separate homeologs by fingerprinting. BES compared to WGS traces showed polyploid-like regions with less than 1% sequence divergence exist at 2.3% of the locations assayed. Conclusion The use of HSVs like SNPs and SNIs to characterize BACs wil improve contig building methods. The implications for bioinformatic and functional annotation of polyploid and paleopolyploid genomes show that a combined approach of BAC fingerprint based physical maps, WGS sequence and HSV-based partitioning of BAC clones from homeologous regions to separate contigs will allow reliable de

  13. Laser microprobe mass analysis (LAMMA) of aluminum and lead in fine roots and their ectomycorrhizal mantles of Norway spruce (Picea abies (L.) Karst.).

    Science.gov (United States)

    Eeckhaoudt, S; Vandeputte, D; Van Praag, H; Van Grieken, R; Jacob, W

    1992-03-01

    Fine roots and ectomycorrhizal root tips were sampled in a Norway spruce (Picea abies (L.) Karst.) stand in the eastern part of the Belgian Ardennes. The cellular and partly subcellular localizations of aluminum and lead were identified by the micro-analytical laser microprobe mass analysis (LAMMA) technique. In fine roots with secondary structure, localization of aluminum was limited to the peripheral cell layers. Lead was found in the outer layers, and also in the primary phloem. Aluminum penetrated the mycorrhizal mantle, but lead was seldom detected in ectomycorrhizae.

  14. Evaluation of BacT/Alert 3D Liquid Culture System for Recovery of Mycobacteria from Clinical Specimens Using Sodium Dodecyl (Lauryl) Sulfate-NaOH Decontamination

    Science.gov (United States)

    Carricajo, A.; Fonsale, N.; Vautrin, A. C.; Aubert, G.

    2001-01-01

    A total of 52 mycobacterial isolates were recovered from 1,197 clinical specimens decontaminated by a sodium dodecyl (lauryl) sulfate (SDS)-NaOH protocol. Of these, 94% were recovered with the BacT/Alert 3D system (Organon Teknika, Durham, N.C.) and 79% were recovered on Löwenstein-Jensen (LJ) medium. Mean times to detection of organisms of the Mycobacterium tuberculosis complex (n = 47) were 22.8 days with LJ medium and 16.2 days with the system. The BacT/Alert 3D system is a rapid and efficient detection system which can be used with an SDS-NaOH decontamination procedure. PMID:11574623

  15. A review of reagents for fluorescence microscopy of cellular compartments and structures, part I: BacMam labeling and reagents for vesicular structures.

    Science.gov (United States)

    Dolman, Nick J; Kilgore, Jason A; Davidson, Michael W

    2013-07-01

    Fluorescent labeling of vesicular structures in cultured cells, particularly for live cells, can be challenging for a number of reasons. The first challenge is to identify a reagent that will be specific enough where some structures have a number of potential reagents and others very few options. The emergence of BacMam constructs has allowed more easy-to-use choices. Presented here is a discussion of BacMam constructs as well as a review of commercially-available reagents for labeling vesicular structures in cells, including endosomes, peroxisomes, lysosomes, and autophagosomes, complete with a featured reagent for each structure, recommended protocol, troubleshooting guide, and example image. © 2013 by John Wiley & Sons, Inc.

  16. A first survey of the rye (Secale cereale) genome composition through BAC end sequencing of the short arm of chromosome 1R

    Czech Academy of Sciences Publication Activity Database

    Bartoš, Jan; Paux, E.; Kofler, R.; Havránková, Miroslava; Kopecký, David; Suchánková, Pavla; Šafář, Jan; Šimková, Hana; Town, C.D.; Lelley, T.; Feuillet, C.; Doležel, Jaroslav

    2008-01-01

    Roč. 8, č. 95 (2008), s. 1-12 ISSN 1471-2229 R&D Projects: GA ČR GP521/06/P412; GA ČR GD521/05/H013; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : BAC libraries * flow-sorted chromosomes Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.030, year: 2008

  17. Sequence-specific DNA binding activity of the cross-brace zinc finger motif of the piggyBac transposase

    Science.gov (United States)

    Morellet, Nelly; Li, Xianghong; Wieninger, Silke A; Taylor, Jennifer L; Bischerour, Julien; Moriau, Séverine; Lescop, Ewen; Bardiaux, Benjamin; Mathy, Nathalie; Assrir, Nadine; Bétermier, Mireille; Nilges, Michael; Hickman, Alison B; Dyda, Fred; Craig, Nancy L; Guittet, Eric

    2018-01-01

    Abstract The piggyBac transposase (PB) is distinguished by its activity and utility in genome engineering, especially in humans where it has highly promising therapeutic potential. Little is known, however, about the structure–function relationships of the different domains of PB. Here, we demonstrate in vitro and in vivo that its C-terminal Cysteine-Rich Domain (CRD) is essential for DNA breakage, joining and transposition and that it binds to specific DNA sequences in the left and right transposon ends, and to an additional unexpectedly internal site at the left end. Using NMR, we show that the CRD adopts the specific fold of the cross-brace zinc finger protein family. We determine the interaction interfaces between the CRD and its target, the 5′-TGCGT-3′/3′-ACGCA-5′ motifs found in the left, left internal and right transposon ends, and use NMR results to propose docking models for the complex, which are consistent with our site-directed mutagenesis data. Our results provide support for a model of the PB/DNA interactions in the context of the transpososome, which will be useful for the rational design of PB mutants with increased activity. PMID:29385532

  18. Tree growth and competition in an old-growth Picea abies forest of boreal Sweden: influence of tree spatial patterning

    Science.gov (United States)

    Fraver, Shawn; D'Amato, Anthony W.; Bradford, John B.; Jonsson, Bengt Gunnar; Jönsson, Mari; Esseen, Per-Anders

    2013-01-01

    Question: What factors best characterize tree competitive environments in this structurally diverse old-growth forest, and do these factors vary spatially within and among stands? Location: Old-growth Picea abies forest of boreal Sweden. Methods: Using long-term, mapped permanent plot data augmented with dendrochronological analyses, we evaluated the effect of neighbourhood competition on focal tree growth by means of standard competition indices, each modified to include various metrics of trees size, neighbour mortality weighting (for neighbours that died during the inventory period), and within-neighbourhood tree clustering. Candidate models were evaluated using mixed-model linear regression analyses, with mean basal area increment as the response variable. We then analysed stand-level spatial patterns of competition indices and growth rates (via kriging) to determine if the relationship between these patterns could further elucidate factors influencing tree growth. Results: Inter-tree competition clearly affected growth rates, with crown volume being the size metric most strongly influencing the neighbourhood competitive environment. Including neighbour tree mortality weightings in models only slightly improved descriptions of competitive interactions. Although the within-neighbourhood clustering index did not improve model predictions, competition intensity was influenced by the underlying stand-level tree spatial arrangement: stand-level clustering locally intensified competition and reduced tree growth, whereas in the absence of such clustering, inter-tree competition played a lesser role in constraining tree growth. Conclusions: Our findings demonstrate that competition continues to influence forest processes and structures in an old-growth system that has not experienced major disturbances for at least two centuries. The finding that the underlying tree spatial pattern influenced the competitive environment suggests caution in interpreting traditional tree

  19. Monitoring intra-annual dynamics of wood formation with microcores and dendrometers in Picea abies at two different altitudes.

    Science.gov (United States)

    Cocozza, Claudia; Palombo, Caterina; Tognetti, Roberto; La Porta, Nicola; Anichini, Monica; Giovannelli, Alessio; Emiliani, Giovanni

    2016-07-01

    Seasonal analyses of cambial cell production and day-by-day stem radial increment can help to elucidate how climate modulates wood formation in conifers. Intra-annual dynamics of wood formation were determined with microcores and dendrometers and related to climatic signals in Norway spruce (Picea abies (L.) Karst.). The seasonal dynamics of these processes were observed at two sites of different altitude, Savignano (650 m a.s.l.) and Lavazè (1800 m a.s.l.) in the Italian Alps. Seasonal dynamics of cambial activity were found to be site specific, indicating that the phenology of cambial cell production is highly variable and plastic with altitude. There was a site-specific trend in the number of cells in the wall thickening phase, with the maximum cell production in early July (DOY 186) at Savignano and in mid-July (DOY 200) at Lavazè. The formation of mature cells showed similar trends at the two sites, although different numbers of cells and timing of cell differentiation were visible in the model shapes; at the end of ring formation in 2010, the number of cells was four times higher at Savignano (106.5 cells) than at Lavazè (26.5 cells). At low altitudes, microcores and dendrometers described the radial growth patterns comparably, though the dendrometer function underlined the higher upper asymptote of maximum growth in comparison with the cell production function. In contrast, at high altitude, these functions exhibited different trends. The best model was obtained by fitting functions of the Gompertz model to the experimental data. By combining radial growth and cambial activity indices we defined a model system able to synchronize these processes. Processes of adaptation of the pattern of xylogenesis occurred, enabling P. abies to occupy sites with contrasting climatic conditions. The use of daily climatic variables in combination with plant functional traits obtained by sensors and/or destructive sampling could provide a suitable tool to better

  20. A PiggyBac mediated approach for lactoferricin gene transfer in bovine mammary epithelial stem cells for management of bovine mastitis.

    Science.gov (United States)

    Sharma, Neelesh; Huynh, Do Luong; Kim, Sung Woo; Ghosh, Mrinmoy; Sodhi, Simrinder Singh; Singh, Amit Kumar; Kim, Nam Eun; Lee, Sung Jin; Hussain, Kafil; Oh, Sung Jong; Jeong, Dong Kee

    2017-11-28

    The antibacterial and anti-inflammatory properties of lactoferricin have been ascribed to its ability to sequester essential iron. The objective of the study was to clone bovine lactoferricin ( LFcinB ) gene into PiggyBac Transposon vector, expression study in the bovine mammary epithelial stem cells (bMESCs) and also to determine the antimicrobial property of recombinant LFcinB against bovine mastitis-causing organisms. The PiggyBac-LFcinB was transfected into bMESCs by electroporation and a three fold of LFcinB secretion was observed in the transfected bMESCs medium by ELISA assay. Furthermore, the assessment of antimicrobial activity against mastitis causing pathogens Staphylococcus aureus and Escherichia coli demonstrated convincing evidence to prove strong antibacterial activity of LFcinB with 14.0±1.0 mm and 18.0±1.5 mm zone of inhibition against both organisms, respectively. The present study provides the convincing evidence to suggest the potential of PiggyBac transposon system to transfer antibacterial peptide into bMESCs or cow mammary gland and also pave the way to use bovine mammary gland as the bioreactors. Simultaneously, it also suggest toward commercial utilization of LFcinB bioreactor system in pharmaceutical industry.

  1. An all-in-one, Tet-On 3G inducible PiggyBac system for human pluripotent stem cells and derivatives.

    Science.gov (United States)

    Randolph, Lauren N; Bao, Xiaoping; Zhou, Chikai; Lian, Xiaojun

    2017-05-08

    Human pluripotent stem cells (hPSCs) offer tremendous promise in tissue engineering and cell-based therapies due to their unique combination of two properties: pluripotency and unlimited proliferative capacity. However, directed differentiation of hPSCs to clinically relevant cell lineages is needed to achieve the goal of hPSC-based therapies. This requires a deep understanding of how cell signaling pathways converge on the nucleus to control differentiation and the ability to dissect gene function in a temporal manner. Here, we report the use of the PiggyBac transposon and a Tet-On 3G drug-inducible gene expression system to achieve versatile inducible gene expression in hPSC lines. Our new system, XLone, offers improvement over previous Tet-On systems with significantly reduced background expression and increased sensitivity to doxycycline. Transgene expression in hPSCs is tightly regulated in response to doxycycline treatment. In addition, the PiggyBac elements in our XLone construct provide a rapid and efficient strategy for generating stable transgenic hPSCs. Our inducible gene expression PiggyBac transposon system should facilitate the study of gene function and directed differentiation in human stem cells.

  2. Time-to-detection of bacteria and yeast with the BACTEC FX versus BacT/Alert Virtuo blood culture systems.

    Science.gov (United States)

    Somily, Ali Mohammed; Habib, Hanan Ahmed; Torchyan, Armen Albert; Sayyed, Samina B; Absar, Muhammed; Al-Aqeel, Rima; Binkhamis, A Khalifa

    2018-01-01

    Bloodstream infections are associated with high rates of morbidity and mortality. Rapid detection of bloodstream infections is important in achieving better patient outcomes. Compare the time-to-detection (TTD) of the new BacT/Alert Virtuo and the BACTEC FX automated blood culture systems. Prospective simulated comparison of two instruments using seeded samples. Medical microbiology laboratory. Blood culture bottles were seeded in triplicate with each of the standard ATCC strains of aerobes, anaerobes and yeast. TTD was calculated as the length of time from the beginning of culture incubation to the detection of bacterial growth. TTD for the various tested organisms on the two microbial detection systems. The 99 bottles of seeded blood cultures incubated in each of the blood culture systems included 21 anaerobic, 39 aerobic and 39 pediatric bottles. The BacT/Alert Virtuo system exhibited significantly shorter TTD for 72.7 % of the tested organisms compared to BACTEC FX system with a median difference in mean TTD of 2.1 hours (interquartile range: 1.5-3.5 hours). The BACTEC FX system was faster in 15.2% (5/33) of microorganisms, with a median difference in mean TTD of 25.9 hours (IQR: 9.1-29.2 hours). TTD was significantly shorter for most of the microorganisms tested on the new BacT/Alert Virtuo system compared to the BACTEC FX system. Use of simulated cultures to assess TTD may not precisely represent clinical blood cultures. None.

  3. De novo 454 sequencing of barcoded BAC pools for comprehensive gene survey and genome analysis in the complex genome of barley

    Directory of Open Access Journals (Sweden)

    Scholz Uwe

    2009-11-01

    Full Text Available Abstract Background De novo sequencing the entire genome of a large complex plant genome like the one of barley (Hordeum vulgare L. is a major challenge both in terms of experimental feasibility and costs. The emergence and breathtaking progress of next generation sequencing technologies has put this goal into focus and a clone based strategy combined with the 454/Roche technology is conceivable. Results To test the feasibility, we sequenced 91 barcoded, pooled, gene containing barley BACs using the GS FLX platform and assembled the sequences under iterative change of parameters. The BAC assemblies were characterized by N50 of ~50 kb (N80 ~31 kb, N90 ~21 kb and a Q40 of 94%. For ~80% of the clones, the best assemblies consisted of less than 10 contigs at 24-fold mean sequence coverage. Moreover we show that gene containing regions seem to assemble completely and uninterrupted thus making the approach suitable for detecting complete and positionally anchored genes. By comparing the assemblies of four clones to their complete reference sequences generated by the Sanger method, we evaluated the distribution, quality and representativeness of the 454 sequences as well as the consistency and reliability of the assemblies. Conclusion The described multiplex 454 sequencing of barcoded BACs leads to sequence consensi highly representative for the clones. Assemblies are correct for the majority of contigs. Though the resolution of complex repetitive structures requires additional experimental efforts, our approach paves the way for a clone based strategy of sequencing the barley genome.

  4. Effect of 3 years' free-air exposure to elevated ozone on mature Norway spruce (Picea abies (L.) Karst.) needle epicuticular wax physicochemical characteristics

    International Nuclear Information System (INIS)

    Percy, Kevin E.; Manninen, Sirkku; Haeberle, Karl-Heinz; Heerdt, C.; Werner, H.; Henderson, Gary W.; Matyssek, Rainer

    2009-01-01

    We examined the effect of ozone (O 3 ) on Norway spruce (Picea abies) needle epicuticular wax over three seasons at the Kranzberg Ozone Fumigation Experiment. Exposure to 2x ambient O 3 ranged from 64.5 to 74.2 μl O 3 l -1 h AOT40, and 117.1 to 123.2 nl O 3 l -1 4th highest daily maximum 8-h average O 3 concentration. The proportion of current-year needle surface covered by wax tubes, tube aggregates, and plates decreased (P = 0.011) under 2x O 3 . Epistomatal chambers had increased deposits of amorphous wax. Proportion of secondary alcohols varied due to year (P = 0.004) and O 3 treatment (P = 0.029). Secondary alcohols were reduced by 9.1% under 2x O 3 . Exposure to 2x O 3 increased (P = 0.037) proportions of fatty acids by 29%. Opposing trends in secondary alcohols and fatty acids indicate a direct action of O 3 on wax biosynthesis. These results demonstrate O 3 -induced changes in biologically important needle surface characteristics of 50-year-old field-grown trees. - Free-air ozone exposure induced changes in needle wax characteristics of mature Picea abies.

  5. Effect of 3 years' free-air exposure to elevated ozone on mature Norway spruce (Picea abies (L.) Karst.) needle epicuticular wax physicochemical characteristics

    Energy Technology Data Exchange (ETDEWEB)

    Percy, Kevin E., E-mail: kpercy@nbnet.nb.c [Natural Resources Canada, Canadian Forest Service-Atlantic Forestry Centre, 1350 Regent Street, Fredericton, NB, E3B 5P7 (Canada); Manninen, Sirkku [Department of Biological and Environmental Sciences, P.O. Box 56, University of Helsinki, 00014 Helsinki (Finland); Department of Biology, P.O. Box 3000, University of Oulu, 90014 Oulu (Finland); Haeberle, Karl-Heinz [Ecophysiology of Plants, Department of Ecology, Technische Universitaet Muenchen, Am Hochanger 13, 85354 Freising (Germany); Heerdt, C.; Werner, H. [Ecoclimatology, Department of Ecology, Technische Universitaet Muenchen, Am Hochanger 13, 85354 Freising (Germany); Henderson, Gary W. [Natural Resources Canada, Canadian Forest Service-Atlantic Forestry Centre, 1350 Regent Street, Fredericton, NB, E3B 5P7 (Canada); Matyssek, Rainer [Ecophysiology of Plants, Department of Ecology, Technische Universitaet Muenchen, Am Hochanger 13, 85354 Freising (Germany)

    2009-05-15

    We examined the effect of ozone (O{sub 3}) on Norway spruce (Picea abies) needle epicuticular wax over three seasons at the Kranzberg Ozone Fumigation Experiment. Exposure to 2x ambient O{sub 3} ranged from 64.5 to 74.2 mul O{sub 3} l{sup -1} h AOT40, and 117.1 to 123.2 nl O{sub 3} l{sup -1} 4th highest daily maximum 8-h average O{sub 3} concentration. The proportion of current-year needle surface covered by wax tubes, tube aggregates, and plates decreased (P = 0.011) under 2x O{sub 3}. Epistomatal chambers had increased deposits of amorphous wax. Proportion of secondary alcohols varied due to year (P = 0.004) and O{sub 3} treatment (P = 0.029). Secondary alcohols were reduced by 9.1% under 2x O{sub 3}. Exposure to 2x O{sub 3} increased (P = 0.037) proportions of fatty acids by 29%. Opposing trends in secondary alcohols and fatty acids indicate a direct action of O{sub 3} on wax biosynthesis. These results demonstrate O{sub 3}-induced changes in biologically important needle surface characteristics of 50-year-old field-grown trees. - Free-air ozone exposure induced changes in needle wax characteristics of mature Picea abies.

  6. Seasonal differences and within-canopy variations of antioxidants in mature spruce (Picea abies) trees under elevated ozone in a free-air exposure system

    International Nuclear Information System (INIS)

    Hofer, Nora; Alexou, Maria; Heerdt, Christian; Loew, Markus; Werner, Herbert; Matyssek, Rainer; Rennenberg, Heinz; Haberer, Kristine

    2008-01-01

    The effect of free-air ozone fumigation and crown position on antioxidants were determined in old-growth spruce (Picea abies) trees in the seasonal course of two consecutive years (2003 and 2004). Levels of total ascorbate and its redox state in the apoplastic washing fluid (AWF) were increased under double ambient ozone concentrations (2 x O 3 ), whilst ascorbate concentrations in needle extracts were unchanged. Concentrations of apoplastic and symplastic ascorbate were significantly higher in 2003 compared to 2004 indicating a combined effect of the drought conditions in 2003 with enhanced ozone exposure. Elevated ozone had only weak effects on total glutathione levels in needle extracts, phloem exudates and xylem saps. Total and oxidised glutathione concentrations were higher in 2004 compared to 2003 and seemed to be more affected by enhanced ozone influx in the more humid year 2004 compared to the combined effect of elevated ozone and drought in 2003 as observed for ascorbate. - Antioxidant defence in sun and shade needles of Picea abies under free-air ozone fumigation in the seasonal course of two consecutive years

  7. Levels of adenine nucleotides (ATP, ADP, AMP) and of inorganic phosphate in needles of Picea abies, representing different stages of development and of pollution dependence

    Energy Technology Data Exchange (ETDEWEB)

    Benz, T; Hampp, R; Horsch, F; Filby, G; Fund, N; Gross, S; Hanisch, B; Kilz, E; Seidel, A [comps.

    1986-04-01

    Levels of adenine nucleotides (ATP, ADP, AMP) and of inorganic phosphate in needles of Picea abies, representing different stages of development and of pollution dependence. Lyophilized needles of Picea abies (Kaelbelescheuer, southern Black Forest) were analyzed for their content of adenine nucleotides (ATP, ADP, AMP: AdN) and of inorganic phosphate (Psub(i)). The metabolite levels were related to needle age, vegetation period and degree of damage (chlorophyll content). The results were as follows: 1) With increasing needle age there is a general decrease in the total AdN-pool. This decrease is most pronounced in very young needles and occurs in both healthy and damaged tissue. 2) The ATP/ADP-ratio of damaged needle is significantly higher than that of healthy ones. 3) Both phosphorylation potential (ATP.(ADP.Psub(i))/sup -1/) and adenylate energy charge ((ATP + 0.5.ADP).(AdN)/sup -1/) are significantly reduced in damaged needles. This is due to relatively higher levels of Psub(i) and of AMP. The results, although incomplete and preliminary, indicate metabolic alterations which have been described for other tissues in response to pollution by photooxidants.

  8. piggyBac transposon somatic mutagenesis with an activated reporter and tracker (PB-SMART for genetic screens in mice.

    Directory of Open Access Journals (Sweden)

    Sean F Landrette

    Full Text Available Somatic forward genetic screens have the power to interrogate thousands of genes in a single animal. Retroviral and transposon mutagenesis systems in mice have been designed and deployed in somatic tissues for surveying hematopoietic and solid tumor formation. In the context of cancer, the ability to visually mark mutant cells would present tremendous advantages for identifying tumor formation, monitoring tumor growth over time, and tracking tumor infiltrations and metastases into wild-type tissues. Furthermore, locating mutant clones is a prerequisite for screening and analyzing most other somatic phenotypes. For this purpose, we developed a system using the piggyBac (PB transposon for somatic mutagenesis with an activated reporter and tracker, called PB-SMART. The PB-SMART mouse genetic screening system can simultaneously induce somatic mutations and mark mutated cells using bioluminescence or fluorescence. The marking of mutant cells enable analyses that are not possible with current somatic mutagenesis systems, such as tracking cell proliferation and tumor growth, detecting tumor cell infiltrations, and reporting tissue mutagenesis levels by a simple ex vivo visual readout. We demonstrate that PB-SMART is highly mutagenic, capable of tumor induction with low copy transposons, which facilitates the mapping and identification of causative insertions. We further integrated a conditional transposase with the PB-SMART system, permitting tissue-specific mutagenesis with a single cross to any available Cre line. Targeting the germline, the system could also be used to conduct F1 screens. With these features, PB-SMART provides an integrated platform for individual investigators to harness the power of somatic mutagenesis and phenotypic screens to decipher the genetic basis of mammalian biology and disease.

  9. A theoretical framework for whole-plant carbon assimilation efficiency based on metabolic scaling theory: a test case using Picea seedlings.

    Science.gov (United States)

    Wang, Zhiqiang; Ji, Mingfei; Deng, Jianming; Milne, Richard I; Ran, Jinzhi; Zhang, Qiang; Fan, Zhexuan; Zhang, Xiaowei; Li, Jiangtao; Huang, Heng; Cheng, Dongliang; Niklas, Karl J

    2015-06-01

    Simultaneous and accurate measurements of whole-plant instantaneous carbon-use efficiency (ICUE) and annual total carbon-use efficiency (TCUE) are difficult to make, especially for trees. One usually estimates ICUE based on the net photosynthetic rate or the assumed proportional relationship between growth efficiency and ICUE. However, thus far, protocols for easily estimating annual TCUE remain problematic. Here, we present a theoretical framework (based on the metabolic scaling theory) to predict whole-plant annual TCUE by directly measuring instantaneous net photosynthetic and respiratory rates. This framework makes four predictions, which were evaluated empirically using seedlings of nine Picea taxa: (i) the flux rates of CO(2) and energy will scale isometrically as a function of plant size, (ii) whole-plant net and gross photosynthetic rates and the net primary productivity will scale isometrically with respect to total leaf mass, (iii) these scaling relationships will be independent of ambient temperature and humidity fluctuations (as measured within an experimental chamber) regardless of the instantaneous net photosynthetic rate or dark respiratory rate, or overall growth rate and (iv) TCUE will scale isometrically with respect to instantaneous efficiency of carbon use (i.e., the latter can be used to predict the former) across diverse species. These predictions were experimentally verified. We also found that the ranking of the nine taxa based on net photosynthetic rates differed from ranking based on either ICUE or TCUE. In addition, the absolute values of ICUE and TCUE significantly differed among the nine taxa, with both ICUE and temperature-corrected ICUE being highest for Picea abies and lowest for Picea schrenkiana. Nevertheless, the data are consistent with the predictions of our general theoretical framework, which can be used to access annual carbon-use efficiency of different species at the level of an individual plant based on simple, direct

  10. Development of fog exposure chambers for field experiments under analytically controlled conditions in order to investigate the effect of hydrogen-peroxide-containing fog on spruces (Picea abies Karst. ). Entwicklung von Benebelungskammern fuer Freilandexperimente unter analytisch kontrollierten Bedingungen zur Untersuchung der Wirkung von wasserstoffperoxidhaltigem Nebel auf Fichten (Picea abies Karst. )

    Energy Technology Data Exchange (ETDEWEB)

    Duelme, W.

    1990-01-12

    This thesis investigated the effect of hydrogen-peroxide-containing fog on spruces (Picea abies Karst.) in a field experiment. Under the influence of hydrogen-peroxide-containing fog, primary needles show an increase in wax contents. This entails reduced water loss through cuticular transpiration and premature aging of needles. The pigment contents of the spruce needles reveal an increase in the contents of xanthophylls and chlorophyll B. Especially the increase in oxygenated carotinoids must be interpreted as an indication of advanced senescence. Regarding phenols, too, changes in the composition of individual classes of substances were established. These point to a shift in biosynthesis in favour of higher phenols. (orig./MG)

  11. Gradients of the content of photosynthetic pigments and radiation as manifestations of the health condition of Norway spruce (Picea abies [L.] Karst.)

    International Nuclear Information System (INIS)

    Zemánek, T.; Martinková, M.; Štěrbová, D.

    2004-01-01

    The distribution and quantities of chlorophyll a + b and carotenoids were determined in a sample tree of Picea abies in order to evaluate its health condition. The content of photosynthetic pigments (PhP) was determined by spectrophotometric analysis. The sample tree was situated at the Rajec nad Svitavou locality, the Drahany Upland, Czech Republic (altitude 625 to 640 m). It was shown that the inner coordination of the content of PhP in the crown in relation to the age of needles and their insolation was sufficient. Thus, the tree did not show impaired health condition and its growth retardation resulted from the short crown. The extent of the photosynthetic apparatus and stability of the tree would be increased particularly after elongation of the lower part of a crown, the so-called compensating part

  12. Gradients of the content of photosynthetic pigments and radiation as manifestations of the health condition of Norway spruce (Picea abies [L.] Karst.)

    International Nuclear Information System (INIS)

    Zemánek, T.; Martinková, M.; Štěrbová, D.

    2004-01-01

    The distribution and quantities of chlorophyll a + b and carotenoids were determined in a sample tree of Picea abies in order to evaluate its health condition. The content of photosynthetic pigments (PhP) was determined by spectrophotometric analysis. The sample tree was situated at the Rajec nad Svitavou locality, the Drahany Upland, Czech Republic (altitude 625 to 640 m). It was shown that the inner coordination of the content of PhP in the crown in relation to the age of needles and their insolation was sufficient. Thus, the tree did not show impaired health condition and its growth retardation resulted from the short crown. The extent of the photosynthetic apparatus and stability of the tree would be increased particularly after elongation of the lower part of a crown, the so-called compensating part. (author)

  13. The impact of long-term CO2 enrichment on sun and shade needles of Norway spruce (Picea abies): Photosynthetic performance, needle anatomy and phenolics accumulation

    Czech Academy of Sciences Publication Activity Database

    Lhotáková, Z.; Urban, Otmar; Dubánková, M.; Cvikrová, Milena; Tomášková, Ivana; Kubínová, Lucie; Zvára, K.; Marek, Michal V.; Albrechtová, J.

    188-189, JUN (2012), s. 60-70 ISSN 0168-9452 R&D Projects: GA ČR(CZ) GAP501/10/0340; GA AV ČR(CZ) IAA600110507; GA AV ČR IAA600870701; GA MŠk(CZ) LC06063; GA MŠk(CZ) LM2010007 Grant - others:GA ČR(CZ) GD206/03/H137 Program:GD Institutional research plan: CEZ:AV0Z60870520; CEZ:AV0Z50380511; CEZ:AV0Z50110509 Keywords : Elevated CO2 * Mesophyll structure * Phenolic compounds * Photosynthesis * Picea abies * Stereological methods Subject RIV: EH - Ecology, Behaviour; ED - Physiology (FGU-C); EF - Botanics (UEB-Q) Impact factor: 2.922, year: 2012

  14. Comparative study on the turnover of quinic- and shikimic acid and of its derivatives in needles of Norway spruce, Picea abies (L.) Karst., affected by 'Waldsterben' syndrome

    International Nuclear Information System (INIS)

    Dittrich, P.; Senser, M.; Frielinghaus, J.

    1989-01-01

    A comparative study on the incorporation of 14 C into quinic- and shikimic acid in spruce needles was carried out with four different syndroms of Picea abies (L.) Karst: a tree from high altitudes of the Bayerischer Wald with the syndrom of 'acute yellowing', a light-sensitive mutant with yellowish needles (Picea abies Karst. finedonensis), a tree exhibiting 'Lametta' syndrome with a translucent crown and twigs hanging down like tinsel, and a healthy tree for control. The needles of these trees were labelled via photosynthetic fixation of 14 CO 2 in June, and the fate of labelled compounds was monitored over 125 days. In the youngest needles incorporation rates of quinic and shikimic acid reached a dominating level of about 60% of the label of soluble metabolites, which underlines the decisive role of both acids during needle development. The yellow mutant 'finedonensis' and the tree with 'Lametta' syndrome, which exhibit rather identical turnover kinetics of quinate and shikimate, show lower rates and reach, in particular in one- and two-year-old needles, only about 50% of the incorporation rates of the control spruce. The tree with the syndrome of 'acute yellowing' exhibits a significant reduction of incorporation already in the youngest needles; the older needles of this tree virtually suspend metabolism of both acids all together, though apparently high but dormant pools of shikimate and quinate are present. The degree of label incorporation into shikimate and quinate may possibly serve as a measure of needle damage. Exclusively in the yellow mutant a novel spruce constituent, 3-0-p-coumaroylquinic acid, could be detected; a related compound, 3-0-p-caffeoylshikimic acid was identified in the needles of the 'acute yellowing' tree. (orig./MG) [de

  15. Some observations on age relationships in spruce-fir regeneration

    Science.gov (United States)

    Barton M. Blum

    1973-01-01

    Measurement of the ages of seedlings of balsam fir (Abies balsamea (L) Mill.), red spruce (Picea rubens Sarg.), and white spruce (Picea glauca (Moench) Voss) 15 years after the first harvest of a two-cut shelterwood operation revealed that very few potential crop-tree seedlings in the sample occurred as advance...

  16. Structure and resilience of fungal communities in Alaskan boreal forest soils

    Science.gov (United States)

    D. Lee Taylor; Ian C. Herriott; Kelsie E. Stone; Jack W. McFarland; Michael G. Booth; Mary Beth Leigh

    2010-01-01

    This paper outlines molecular analyses of soil fungi within the Bonanza Creek Long Term Ecological Research program. We examined community structure in three studies in mixed upland, black spruce (Picea mariana (Mill.) BSP), and white spruce (Picea glauca (Moench) Voss) forests and examined taxa involved in cellulose...

  17. Dicty_cDB: CHD758 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available nce. 38 0.29 2 CO488299 |CO488299.1 GQ02511.B3_K04 GQ025: Annual flush SHOOTS diurnal cycle- trees Picea gla...511.B3_O14 GQ025: Annual flush SHOOTS diurnal cycle- trees Picea glauca cDNA clone GQ02511_O14 5', mRNA sequ

  18. Etude du risque d'inondation en aval du delta du fleuve rouge en utilisant la teledetection et les sig: Le cas du district de Bac Hung Hai

    Science.gov (United States)

    Bui, Duc Viet

    The Bac Hung Hai zone is the greatest basin in the Red River Delta in Vietnam and also one of the most densely populated regions of the planet. It is mainly a rural region and its economy is dominated by agriculture. In the context of frequent and larger floods in the Bac Hung Hai zone, causing deep socio-economical consequences, the focus of this study is to establish cartography of the high risk areas for flooding in the Bac Hung Hai region using remote sensing and GIS to assist land management. The preparation of a map describing land management in this region is more complicated because parcels for farming are very small and not homogeneous. A consistent and precise map of land use is essential for studies of flooding. The secondary objective is to improve the land use map. To this effect, a classification has been applied to the combination of the spectral bands and textures (TM and ETM+) of Landsat and a radar image (ERS). The addition of this information to the spectral bands increases the accuracy of classification by 1% to 4%, according to the dates selected. Additionally, in the study zone where there are few days without clouds, a problem related to the optical satellite image is the cloud cover. Then, the use of radar images will provide ground information for areas hidden by clouds where spectral images are not sufficient. To reach these goals, we have determined the main biophysical considerations that influence flooding. Then, these considerations have been combined in a multi-criteria analysis to evaluate the risks of flooding in the entire basin area. The results show that high to very high risks affect 47% of the area studied and that the south-east region, center, and north-east present the greatest risk. Keywords. Flood risks, remote sensing, GIS, land use, multicriteria analysis, Red river delta, Vietnam.

  19. Heterogeneous transgene expression in the retinas of the TH-RFP, TH-Cre, TH-BAC-Cre and DAT-Cre mouse lines.

    Science.gov (United States)

    Vuong, H E; Pérez de Sevilla Müller, L; Hardi, C N; McMahon, D G; Brecha, N C

    2015-10-29

    Transgenic mouse lines are essential tools for understanding the connectivity, physiology and function of neuronal circuits, including those in the retina. This report compares transgene expression in the retina of a tyrosine hydroxylase (TH)-red fluorescent protein (RFP) mouse line with three catecholamine-related Cre recombinase mouse lines [TH-bacterial artificial chromosome (BAC)-, TH-, and dopamine transporter (DAT)-Cre] that were crossed with a ROSA26-tdTomato reporter line. Retinas were evaluated and immunostained with commonly used antibodies including those directed to TH, GABA and glycine to characterize the RFP or tdTomato fluorescent-labeled amacrine cells, and an antibody directed to RNA-binding protein with multiple splicing to identify ganglion cells. In TH-RFP retinas, types 1 and 2 dopamine (DA) amacrine cells were identified by their characteristic cellular morphology and type 1 DA cells by their expression of TH immunoreactivity. In the TH-BAC-, TH-, and DAT-tdTomato retinas, less than 1%, ∼ 6%, and 0%, respectively, of the fluorescent cells were the expected type 1 DA amacrine cells. Instead, in the TH-BAC-tdTomato retinas, fluorescently labeled AII amacrine cells were predominant, with some medium diameter ganglion cells. In TH-tdTomato retinas, fluorescence was in multiple neurochemical amacrine cell types, including four types of polyaxonal amacrine cells. In DAT-tdTomato retinas, fluorescence was in GABA immunoreactive amacrine cells, including two types of bistratified and two types of monostratified amacrine cells. Although each of the Cre lines was generated with the intent to specifically label DA cells, our findings show a cellular diversity in Cre expression in the adult retina and indicate the importance of careful characterization of transgene labeling patterns. These mouse lines with their distinctive cellular labeling patterns will be useful tools for future studies of retinal function and visual processing. Published by Elsevier

  20. Evaluation of shrub and tree species used for revegetating copper mine wastes in the south-western United States. [Dodonea viscosa (L. ) Jacq. , Baccharis sarothroides Gray, Cerdicium microphyllum Torr. , and Nicotiana glauca Grah

    Energy Technology Data Exchange (ETDEWEB)

    Norem, M A; Day, A D; Ludeke, K L

    1982-12-01

    The revegetation work begun in 1970 at Cyprus Pima Mine, an open pit copper mine south-west of Tucson, Arizona, was evaluated to determine the effects of slope aspect and mining waste material on plant survival and growth. Only one shrub, Dodonea viscosa (L.) Jacq. (hopbush), survived on the east slope. Baccharis sarothroides Gray (desert broom) was prolific on the north aspect. Cerdicium microphyllum Torr. (palo verde) survived best on the east slope and Nicotiana glauca Grah. (desert tobacco) survived only on the north slope. The survival of the other tree species was not affected by slope aspect. Slope exposure did not affect tree size, except for Eucalyptus microtheca Muell. (tiny capsule eucalyptus) which grew larger on the north slope. E. rostrata Schlechtend (red gum eucalyptus) grew taller in overburden than in tailing on the east slope. Differences in soil material within each slope exposure did not significantly affect growth of the other tree species. In semi-arid regions, such as south-western Arizona, even slight differences in available moisture may determine the survival of a plant species in the area. (Refs. 11).

  1. Melanomacrophages in three species of free-ranging sharks from the northwestern Atlantic, the blue shark Prionacae glauca (L.), the shortfin mako, Isurus oxyrhinchus Rafinesque, and the thresher, Alopias vulpinus (Bonnaterre).

    Science.gov (United States)

    Borucinska, J D; Kotran, K; Shackett, M; Barker, T

    2009-10-01

    The melanomacrophage aggregates or cells (MMC) are commonly used as biomarkers of exposure to pollution in fish, albeit their numbers and morphological characteristics can be influenced not only by environmental toxins but also by a range of physiological parameters and pathological conditions. Accordingly, before we can use MMC as biomarkers in any fish species, their normal, 'background' characteristics have to be established in apparently healthy fish. The knowledge of MMC in sharks is minimal. The aim of this study was to characterize MMC from 51 free-ranging, large pelagic sharks from the northwestern Atlantic, including shortfin mako, Isurus oxyrhinchus, thresher, Alopias vulpinus and blue shark, Prionacae glauca. The rationale of this study was twofold. First, because, sharks have life histories predisposing them to the accumulation of environmental toxins they constitute good sentinel species of the health of the global marine ecosystem. Second, because many shark populations are in decline or threatened by extinction, we need to expand our understanding of their health status in order to develop appropriate protective measures. All sharks were collected by sports fishing gear between June and August in 2007. Their health condition was assessed by necropsy, morphometrics, and by microscopic examination of gonads, livers, spleens and kidneys. Routine, haematoxylin and eosin and/or Pearl's reagent-stained paraffin embedded sections were studied by light microscopy. Our results provide the first data on the morphometric and morphological characteristics of MMC in viscera of apparently healthy free-ranging sharks from the northwestern Atlantic.

  2. Sexual violence and the risk of HIV transmission in sexual partners of male injecting drug users in Tien Du district, Bac Ninh province of Vietnam.

    Science.gov (United States)

    Do, Vinh Thi; Ho, Hien Thi; Nguyen, Tri Manh; Do, Huynh Khac

    2018-04-01

    We conducted a cross-sectional study among 148 women who were regular sexual partners of male injecting drug users in Tien Du, Bac Ninh province, Vietnam to identify the rate of HIV infection and factors associated with HIV transmission among them. HIV infection rate among sexual partners was high, 11.5%. Sexual violence was prevalent, 63.5% among sexual partners; 94.1% (16/17) among those with HIV. We discovered an association between sexual violence and HIV infection. Sexual partners suffering from sexual violence caused by their regular sexual partners faced 9.24 times higher HIV risk than those who did not have sexual violence.

  3. A novel resource for genomics of Triticeae: BAC library specific for the short arm of rye (Secale cereale L.) chromosome 1R (1RS)

    Czech Academy of Sciences Publication Activity Database

    Šimková, Hana; Šafář, Jan; Suchánková, Pavla; Kovářová, Pavlína; Bartoš, Jan; Kubaláková, Marie; Janda, Jaroslav; Čihalíková, Jarmila; Mago, R.; Lelley, T.; Doležel, Jaroslav

    2008-01-01

    Roč. 9, č. 237 (2008), s. 101-109 ISSN 1471-2164 R&D Projects: GA ČR GA521/04/0607; GA ČR GP521/05/P257; GA ČR GD521/05/H013; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50380511 Keywords : flow cytometry * flow-sorted chromosomes * BAC library Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.926, year: 2008

  4. Construction and characterization of two BAC libraries representing a deep-coverage of the genome of chicory (Cichorium intybus L., Asteraceae

    Directory of Open Access Journals (Sweden)

    Gonthier Lucy

    2010-08-01

    Full Text Available Abstract Background The Asteraceae represents an important plant family with respect to the numbers of species present in the wild and used by man. Nonetheless, genomic resources for Asteraceae species are relatively underdeveloped, hampering within species genetic studies as well as comparative genomics studies at the family level. So far, six BAC libraries have been described for the main crops of the family, i.e. lettuce and sunflower. Here we present the characterization of BAC libraries of chicory (Cichorium intybus L. constructed from two genotypes differing in traits related to sexual and vegetative reproduction. Resolving the molecular mechanisms underlying traits controlling the reproductive system of chicory is a key determinant for hybrid development, and more generally will provide new insights into these traits, which are poorly investigated so far at the molecular level in Asteraceae. Findings Two bacterial artificial chromosome (BAC libraries, CinS2S2 and CinS1S4, were constructed from HindIII-digested high molecular weight DNA of the contrasting genotypes C15 and C30.01, respectively. C15 was hermaphrodite, non-embryogenic, and S2S2 for the S-locus implicated in self-incompatibility, whereas C30.01 was male sterile, embryogenic, and S1S4. The CinS2S2 and CinS1S4 libraries contain 89,088 and 81,408 clones. Mean insert sizes of the CinS2S2 and CinS1S4 clones are 90 and 120 kb, respectively, and provide together a coverage of 12.3 haploid genome equivalents. Contamination with mitochondrial and chloroplast DNA sequences was evaluated with four mitochondrial and four chloroplast specific probes, and was estimated to be 0.024% and 1.00% for the CinS2S2 library, and 0.028% and 2.35% for the CinS1S4 library. Using two single copy genes putatively implicated in somatic embryogenesis, screening of both libraries resulted in detection of 12 and 13 positive clones for each gene, in accordance with expected numbers. Conclusions This

  5. Construction and characterization of two BAC libraries representing a deep-coverage of the genome of chicory (Cichorium intybus L., Asteraceae).

    Science.gov (United States)

    Gonthier, Lucy; Bellec, Arnaud; Blassiau, Christelle; Prat, Elisa; Helmstetter, Nicolas; Rambaud, Caroline; Huss, Brigitte; Hendriks, Theo; Bergès, Hélène; Quillet, Marie-Christine

    2010-08-11

    The Asteraceae represents an important plant family with respect to the numbers of species present in the wild and used by man. Nonetheless, genomic resources for Asteraceae species are relatively underdeveloped, hampering within species genetic studies as well as comparative genomics studies at the family level. So far, six BAC libraries have been described for the main crops of the family, i.e. lettuce and sunflower. Here we present the characterization of BAC libraries of chicory (Cichorium intybus L.) constructed from two genotypes differing in traits related to sexual and vegetative reproduction. Resolving the molecular mechanisms underlying traits controlling the reproductive system of chicory is a key determinant for hybrid development, and more generally will provide new insights into these traits, which are poorly investigated so far at the molecular level in Asteraceae. Two bacterial artificial chromosome (BAC) libraries, CinS2S2 and CinS1S4, were constructed from HindIII-digested high molecular weight DNA of the contrasting genotypes C15 and C30.01, respectively. C15 was hermaphrodite, non-embryogenic, and S2S2 for the S-locus implicated in self-incompatibility, whereas C30.01 was male sterile, embryogenic, and S1S4. The CinS2S2 and CinS1S4 libraries contain 89,088 and 81,408 clones. Mean insert sizes of the CinS2S2 and CinS1S4 clones are 90 and 120 kb, respectively, and provide together a coverage of 12.3 haploid genome equivalents. Contamination with mitochondrial and chloroplast DNA sequences was evaluated with four mitochondrial and four chloroplast specific probes, and was estimated to be 0.024% and 1.00% for the CinS2S2 library, and 0.028% and 2.35% for the CinS1S4 library. Using two single copy genes putatively implicated in somatic embryogenesis, screening of both libraries resulted in detection of 12 and 13 positive clones for each gene, in accordance with expected numbers. This indicated that both BAC libraries are valuable tools for molecular

  6. Identification of an extensive gene cluster among a family of PPOs in Trifolium pratense L. (red clover using a large insert BAC library

    Directory of Open Access Journals (Sweden)

    Thomas Ann

    2009-07-01

    Full Text Available Abstract Background Polyphenol oxidase (PPO activity in plants is a trait with potential economic, agricultural and environmental impact. In relation to the food industry, PPO-induced browning causes unacceptable discolouration in fruit and vegetables: from an agriculture perspective, PPO can protect plants against pathogens and environmental stress, improve ruminant growth by increasing nitrogen absorption and decreasing nitrogen loss to the environment through the animal's urine. The high PPO legume, red clover, has a significant economic and environmental role in sustaining low-input organic and conventional farms. Molecular markers for a range of important agricultural traits are being developed for red clover and improved knowledge of PPO genes and their structure will facilitate molecular breeding. Results A bacterial artificial chromosome (BAC library comprising 26,016 BAC clones with an average 135 Kb insert size, was constructed from Trifolium pratense L. (red clover, a diploid legume with a haploid genome size of 440–637 Mb. Library coverage of 6–8 genome equivalents ensured good representation of genes: the library was screened for polyphenol oxidase (PPO genes. Two single copy PPO genes, PPO4 and PPO5, were identified to add to a family of three, previously reported, paralogous genes (PPO1–PPO3. Multiple PPO1 copies were identified and characterised revealing a subfamily comprising three variants PPO1/2, PPO1/4 and PPO1/5. Six PPO genes clustered within the genome: four separate BAC clones could be assembled onto a predicted 190–510 Kb single BAC contig. Conclusion A PPO gene family in red clover resides as a cluster of at least 6 genes. Three of these genes have high homology, suggesting a more recent evolutionary event. This PPO cluster covers a longer region of the genome than clusters detected in rice or previously reported in tomato. Full-length coding sequences from PPO4, PPO5, PPO1/5 and PPO1/4 will facilitate

  7. Conserved synteny of genes between chromosome 15 of Bombyx mori and a chromosome of Manduca sexta shown by five-color BAC-FISH

    Czech Academy of Sciences Publication Activity Database

    Sahara, K.; Yoshido, A.; Marec, František; Vrbová, Iva; Zhang, H. B.; Wu, Ch. C.; Goldsmith, M. R.; Yasukochi, Y.

    2007-01-01

    Roč. 50, č. 11 (2007), s. 1061-1065 ISSN 0831-2796 R&D Projects: GA ČR GA206/06/1860 Grant - others:Japan Society for the Promotion of Science(JP) 18380037; US National Science Foundation(US) IBN0208388 Institutional research plan: CEZ:AV0Z50070508 Source of funding: O - operačné programy ; O - operačné programy Keywords : BAC-FISH * Bombyx mori * chromosome Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.785, year: 2007

  8. A human fecal contamination score for ranking recreational sites using the HF183/BacR287 quantitative real-time PCR method.

    Science.gov (United States)

    Cao, Yiping; Sivaganesan, Mano; Kelty, Catherine A; Wang, Dan; Boehm, Alexandria B; Griffith, John F; Weisberg, Stephen B; Shanks, Orin C

    2018-01-01

    Human fecal pollution of recreational waters remains a public health concern worldwide. As a result, there is a growing interest in the application of human-associated fecal source identification quantitative real-time PCR (qPCR) technologies for water quality research and management. However, there are currently no standardized approaches for field implementation and interpretation of qPCR data. In this study, a standardized HF183/BacR287 qPCR method was combined with a water sampling strategy and a novel Bayesian weighted average approach to establish a human fecal contamination score (HFS) that can be used to prioritize sampling sites for remediation based on measured human waste levels. The HFS was then used to investigate 975 study design scenarios utilizing different combinations of sites with varying sampling intensities (daily to once per week) and number of qPCR replicates per sample (2-14 replicates). Findings demonstrate that site prioritization with HFS is feasible and that both sampling intensity and number of qPCR replicates influence reliability of HFS estimates. The novel data analysis strategy presented here provides a prescribed approach for the implementation and interpretation of human-associated HF183/BacR287 qPCR data with the goal of site prioritization based on human fecal pollution levels. In addition, information is provided for future users to customize study designs for optimal HFS performance. Published by Elsevier Ltd.

  9. Comparison of the chromosome maps around a resistance hot spot on chromosome 5 of potato and tomato using BAC-FISH painting.

    Science.gov (United States)

    Achenbach, Ute C; Tang, Xiaomin; Ballvora, Agim; de Jong, Hans; Gebhardt, Christiane

    2010-02-01

    Potato chromosome 5 harbours numerous genes for important qualitative and quantitative traits, such as resistance to the root cyst nematode Globodera pallida and the late blight fungus, Phytophthora infestans. The genes make up part of a "hot spot" for resistances to various pathogens covering a genetic map length of 3 cM between markers GP21 and GP179. We established the physical size and position of this region on chromosome 5 in potato and tomato using fluorescence in situ hybridization (FISH) on pachytene chromosomes. Five potato bacterial artificial chromosome (BAC) clones with the genetically anchored markers GP21, R1-contig (proximal end), CosA, GP179, and StPto were selected, labeled with different fluorophores, and hybridized in a five-colour FISH experiment. Our results showed the location of the BAC clones in the middle of the long arm of chromosome 5 in both potato and tomato. Based on chromosome measurements, we estimate the physical size of the GP21-GP179 interval at 0.85 Mb and 1.2 Mb in potato and tomato, respectively. The GP21-GP179 interval is part of a genome segment known to have inverted map positions between potato and tomato.

  10. [Use of the recombinant baculovirus BacVP6C for the construction of an internal positive control of rotavirus C].

    Science.gov (United States)

    Abid-Ayadi, I; Guix, S; Pintó, R M; Bosch, A

    2011-06-01

    Unlike group A, a few studies have interested other groups of the rotavirus, especially in Tunisia. The role of rotavirus C (RVC) infection is underestimated because of its sporadic nature. The aim of our study was to develop rapid diagnostic procedures of RVC by using an internal positive control of reverse transcription PCR (RT-PCR). The internal positive control (386pb) was designed from the recombinant baculovirus BacVP6C containing the full length cDNA of the Cowden strain gene 5 (1353pb). A fragment of 596pb was amplified by PCR using the BacVP6C DNA ds as template. Then, a central part of 210pb was deleted and the remaining fragment (386pb) was cloned into pGEM-3Zf(+) plasmid between SP6 and T7 RNA polymerase promoters. The obtained recombinant plasmid "pIAM1" was then used for the generation of the internal positive control by in vitro transcription. The sensibility of the RT-PCR was about 3.66×10(5) molecules of RNA/μl. The use of a shorter positive control, as compared to the wild type, allows increased specificity of the RT-PCR reaction, and could be used for efficient diagnostic and surveillance of RVC-caused diseases. Copyright © 2009 Elsevier Masson SAS. All rights reserved.

  11. Deletion of the budBAC operon in Klebsiella pneumoniae to understand the physiological role of 2,3-butanediol biosynthesis.

    Science.gov (United States)

    Jeong, Daun; Yang, Jeongmo; Lee, Soojin; Kim, Borim; Um, Youngsoon; Kim, Youngrok; Ha, Kyoung-Su; Lee, Jinwon

    2016-05-18

    Klebsiella pneumoniae is known to produce 2,3-butanediol (2,3-BDO), a valuable chemical. In K. pneumoniae, the 2,3-BDO operon (budBAC) is involved in the production of 2,3-BDO. To observe the physiological role of the 2,3-BDO operon in a mixed acid fermentation, we constructed a budBAC-deleted strain (SGSB109). The production of extracellular metabolites, CO2 emission, carbon distribution, and NADH/NAD(+) balance of SGSB109 were compared with the parent strain (SGSB100). When comparing the carbon distribution at 15 hr, four significant differences were observed: in 2,3-BDO biosynthesis, lactate and acetate production (lactate and acetate production increased 2.3-fold and 4.1-fold in SGSB109 compared to SGSB100), CO2 emission (higher in SGSB100), and carbon substrate uptake (higher in SGSB100). Previous studies on the inactivation of the 2,3-BDO operon were focused on the increase of 1,3-propanediol production. Few studies have been done observing the role of 2,3-BDO biosynthesis. This study provides a prime insight into the role of 2,3-BDO biosynthesis of K. pneumoniae.

  12. NATURAL MUTATION IN THE GENE OF RESPONSE REGULATOR BgrR RESULTING IN REPRESSION OF Bac PROTEIN SYNTHESIS, A PATHOGENICITY FACTOR OF STREPTOCOCCUS AGALACTIAE

    Directory of Open Access Journals (Sweden)

    A. S. Rozhdestvenskaya

    2013-01-01

    Full Text Available Abstract. Streptococcus agalactiae can cause variety of diseases of newborns and adults. For successful colonization of different human tissues and organs as well as for suppression of the host immune system S. agalactiae expresses numerous virulence factors. For coordinated expression of the virulence genes S. agalactiae employs regulatory molecules including regulatory proteins of two-component systems. Results of the present study demonstrated that in S. agalactiae strain A49V the natural mutation in the brgR gene encoding for BgrR regulatory protein, which is component of regulatory system BgrRS, resulted in the repression of Bac protein synthesis, a virulence factor of S. agalactiae. A single nucleotide deletion in the bgrR gene has caused a shift of the reading frame and the changes in the primary, secondary and tertiary structures of the BgrR protein. The loss of functional activity of BgrR protein in A49V strain and repression of Bac protein synthesis have increased virulence of the strain in experimental animal streptococcal infection.

  13. nanos-Driven expression of piggyBac transposase induces mobilization of a synthetic autonomous transposon in the malaria vector mosquito, Anopheles stephensi.

    Science.gov (United States)

    Macias, Vanessa M; Jimenez, Alyssa J; Burini-Kojin, Bianca; Pledger, David; Jasinskiene, Nijole; Phong, Celine Hien; Chu, Karen; Fazekas, Aniko; Martin, Kelcie; Marinotti, Osvaldo; James, Anthony A

    2017-08-01

    Transposons are a class of selfish DNA elements that can mobilize within a genome. If mobilization is accompanied by an increase in copy number (replicative transposition), the transposon may sweep through a population until it is fixed in all of its interbreeding members. This introgression has been proposed as the basis for drive systems to move genes with desirable phenotypes into target species. One such application would be to use them to move a gene conferring resistance to malaria parasites throughout a population of vector mosquitos. We assessed the feasibility of using the piggyBac transposon as a gene-drive mechanism to distribute anti-malarial transgenes in populations of the malaria vector, Anopheles stephensi. We designed synthetic gene constructs that express the piggyBac transposase in the female germline using the control DNA of the An. stephensi nanos orthologous gene linked to marker genes to monitor inheritance. Two remobilization events were observed with a frequency of one every 23 generations, a rate far below what would be useful to drive anti-pathogen transgenes into wild mosquito populations. We discuss the possibility of optimizing this system and the impetus to do so. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  14. Efficient, footprint-free human iPSC genome editing by consolidation of Cas9/CRISPR and piggyBac technologies.

    Science.gov (United States)

    Wang, Gang; Yang, Luhan; Grishin, Dennis; Rios, Xavier; Ye, Lillian Y; Hu, Yong; Li, Kai; Zhang, Donghui; Church, George M; Pu, William T

    2017-01-01

    Genome editing of human induced pluripotent stem cells (hiPSCs) offers unprecedented opportunities for in vitro disease modeling and personalized cell replacement therapy. The introduction of Cas9-directed genome editing has expanded adoption of this approach. However, marker-free genome editing using standard protocols remains inefficient, yielding desired targeted alleles at a rate of ∼1-5%. We developed a protocol based on a doxycycline-inducible Cas9 transgene carried on a piggyBac transposon to enable robust and highly efficient Cas9-directed genome editing, so that a parental line can be expeditiously engineered to harbor many separate mutations. Treatment with doxycycline and transfection with guide RNA (gRNA), donor DNA and piggyBac transposase resulted in efficient, targeted genome editing and concurrent scarless transgene excision. Using this approach, in 7 weeks it is possible to efficiently obtain genome-edited clones with minimal off-target mutagenesis and with indel mutation frequencies of 40-50% and homology-directed repair (HDR) frequencies of 10-20%.

  15. The Performance of the Four Anaerobic Blood Culture Bottles BacT/ALERT-FN, -FN Plus, BACTEC-Plus and -Lytic in Detection of Anaerobic Bacteria and Identification by Direct MALDI-TOF MS.

    Directory of Open Access Journals (Sweden)

    Mohammed Almuhayawi

    Full Text Available Detection and identification of anaerobic bacteria in blood cultures (BC is a well-recognized challenge in clinical microbiology. We studied 100 clinical anaerobic BC isolates to evaluate the performance of BacT/ALERT-FN, -FN Plus (BioMérieux, BACTEC-Plus and -Lytic (Becton Dickinson BioSciences BC bottles in detection and time to detection (TTD of anaerobic bacteria. BACTEC Lytic had higher detection rate (94/100, 94% than BacT/ALERT FN Plus (80/100, 80% (p<0.01 in the studied material. There was no significant difference in detection of anaerobic bacteria among the remaining bottle types. The 67 anaerobic bacteria that signalled positive in all four bottle types were analyzed to compare the time to detection (TTD and isolates were directly identified by MALDI-TOF MS. There was a significant difference in TTD among the four bottle types (p<0.0001. The shortest median TTD was 18 h in BACTEC Lytic followed by BacT/ALERT FN (23.5 h, BACTEC Plus (27 h and finally BacT/ALERT FN Plus (38 h bottles. In contrast, MALDI-TOF MS performed similarly in all bottle types with accurate identification in 51/67 (76% BacT/ALERT FN, 51/67 (76% BacT/ALERT FN Plus, 53/67 (79% BACTEC Plus and 50/67 (75% BACTEC Lytic bottles. In conclusion, BACTEC Lytic bottles have significantly better detection rates and shorter TTD compared to the three other bottle types. The anaerobic BC bottles are equally suitable for direct MALDI-TOF MS for rapid and reliable identification of common anaerobic bacteria. Further clinical studies are warranted to investigate the performance of anaerobic BC bottles in detection of anaerobic bacteria and identification by direct MALDI-TOF MS.

  16. Expression of the Azotobacter vinelandii Poly-β-Hydroxybutyrate Biosynthetic phbBAC Operon Is Driven by Two Overlapping Promoters and Is Dependent on the Transcriptional Activator PhbR

    OpenAIRE

    Peralta-Gil, Martín; Segura, Daniel; Guzmán, Josefina; Servín-González, Luis; Espín, Guadalupe

    2002-01-01

    The Azotobacter vinelandii phbBAC genes encode the enzymes for poly-β-hydroxybutyrate (PHB) synthesis. The phbR gene, which is located upstream of and in the opposite direction of phbBAC, encodes PhbR, a transcriptional activator which is a member of the AraC family of activators. Here we report that a mutation in phbR reduced PHB accumulation and transcription of a phbB-lacZ fusion. We also report that phbB is transcribed from two overlapping promoters, pB1 and pB2. The region corresponding ...

  17. Organizace embryí ve shlucích raných somatických embryí smrku ztepilého (Picea abies/L./Karst.)

    Czech Academy of Sciences Publication Activity Database

    Petřek, J.; Havel, L.; Víteček, J.; Vlašínová, H.; Adam, V.; Babula, P.; Bartušek, Karel; Potěšil, D.; Kizek, R.

    2005-01-01

    Roč. 99, č. 5 (2005), s. 377-378 ISSN 0009-2770. [Mezioborové setkání mladých biologů, biochemiků a chemiků /5./. 15.06.2005-18.06.2005, Devět skal - Žďárské vrchy] R&D Projects: GA AV ČR(CZ) IAA2065201 Keywords : somatic embrya * explantate cultivation * Picea abies Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering

  18. The role of Nicotiana gluca Graham (paraguayan herbs as an adjuvant in immunomodulation of Newcastle disease vaccine for broilers Estudo da ação de Nicotiana glauca Graham (erva paraguaia como coadjuvante em vacina contra a doença de Newcastle em frangos de corte

    Directory of Open Access Journals (Sweden)

    Fabiane Pereira Gentilini

    2008-07-01

    Full Text Available The Nicotiana glauca is a native plant species from Argentina, but found all over South América, being used against headaches, rheumatism, injuries, ulcers, and so on. Researchers have considered it as having immunomodulation effect. This study was conducted to investigate the use of a aqueous extract of Nicotiana glauca Graham as an immunomodulator (adjuvant of a Newcastle disease vaccine.. A total of 56 broilers were distributed into 4 experimental groups. Each one of them received 3 dosages of this vaccine with or without the addition of different concentrations of the extract Using hemmoaglutination inhibition techniques , the results have shown differences (P<0.05 in the third sera collection. An increase in the antibody titer with the inclusion of 5 mg/dosage of the extract (Treatment 3 as compared to 1 mg/dosage (Treatment 2 and 10 mg/dosage of the extract (Treatment 4 was observed, However, birds from Treatment 3 did not differ (P> 0.05 from Treatment 1. These results indicated that further investigations are required, including the use of cytotoxicity tests in vitro, to evaluate the immunomodulation effect of this extract.

     

    KEY WORDS: Immunomodulation effect, Nicotiana glauca Graham, vaccine.

    A Nicotiana glauca Graham é uma espécie nativa da Argentina, bem distribuída na América do Sul, sendo empregada, popularmente, contra dores de cabeça, dores reumáticas, cicatrização de feridas e úlceras, entre outros. Pesquisas têm avaliado a sua ação na potencialização da resposta imune. Assim, com este estudo, buscou-se avaliar a ação de um extrato aquoso de Nicotiana glauca Graham como coadjuvante imunológico em uma vacina contra a doença de Newcastle (DNC. Utilizaram-se 56 frangos de corte, distribuídos em quatro grupos experimentais, que receberam tr

  19. How does climate influence xylem morphogenesis over the growing season? Insights from long-term intra-ring anatomy in Picea abies.

    Science.gov (United States)

    Castagneri, Daniele; Fonti, Patrick; von Arx, Georg; Carrer, Marco

    2017-04-01

    During the growing season, the cambium of conifer trees produces successive rows of xylem cells, the tracheids, that sequentially pass through the phases of enlargement and secondary wall thickening before dying and becoming functional. Climate variability can strongly influence the kinetics of morphogenetic processes, eventually affecting tracheid shape and size. This study investigates xylem anatomical structure in the stem of Picea abies to retrospectively infer how, in the long term, climate affects the processes of cell enlargement and wall thickening. Tracheid anatomical traits related to the phases of enlargement (diameter) and wall thickening (wall thickness) were innovatively inspected at the intra-ring level on 87-year-long tree-ring series in Picea abies trees along a 900 m elevation gradient in the Italian Alps. Anatomical traits in ten successive tree-ring sectors were related to daily temperature and precipitation data using running correlations. Close to the altitudinal tree limit, low early-summer temperature negatively affected cell enlargement. At lower elevation, water availability in early summer was positively related to cell diameter. The timing of these relationships shifted forward by about 20 (high elevation) to 40 (low elevation) d from the first to the last tracheids in the ring. Cell wall thickening was affected by climate in a different period in the season. In particular, wall thickness of late-formed tracheids was strongly positively related to August-September temperature at high elevation. Morphogenesis of tracheids sequentially formed in the growing season is influenced by climate conditions in successive periods. The distinct climate impacts on cell enlargement and wall thickening indicate that different morphogenetic mechanisms are responsible for different tracheid traits. Our approach of long-term and high-resolution analysis of xylem anatomy can support and extend short-term xylogenesis observations, and increase our

  20. Molecular control of normal and acrocona mutant seed cone development in Norway spruce (Picea abies) and the evolution of conifer ovule-bearing organs.

    Science.gov (United States)

    Carlsbecker, Annelie; Sundström, Jens F; Englund, Marie; Uddenberg, Daniel; Izquierdo, Liz; Kvarnheden, Anders; Vergara-Silva, Francisco; Engström, Peter

    2013-10-01

    Reproductive organs in seed plants are morphologically divergent and their evolutionary history is often unclear. The mechanisms controlling their development have been extensively studied in angiosperms but are poorly understood in conifers and other gymnosperms. Here, we address the molecular control of seed cone development in Norway spruce, Picea abies. We present expression analyses of five novel MADS-box genes in comparison with previously identified MADS and LEAFY genes at distinct developmental stages. In addition, we have characterized the homeotic transformation from vegetative shoot to female cone and associated changes in regulatory gene expression patterns occurring in the acrocona mutant. The analyses identified genes active at the onset of ovuliferous and ovule development and identified expression patterns marking distinct domains of the ovuliferous scale. The reproductive transformation in acrocona involves the activation of all tested genes normally active in early cone development, except for an AGAMOUS-LIKE6/SEPALLATA (AGL6/SEP) homologue. This absence may be functionally associated with the nondeterminate development of the acrocona ovule-bearing scales. Our morphological and gene expression analyses give support to the hypothesis that the modern cone is a complex structure, and the ovuliferous scale the result of reductions and compactions of an ovule-bearing axillary short shoot in cones of Paleozoic conifers. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

  1. Recovery of photosynthesis in 1-year-old needles of unfertilized and fertilized Norway spruce (Picea abies (L.) Karst.) during spring.

    Science.gov (United States)

    Strand, M; Lundmark, T

    1995-03-01

    Photosynthetic O(2) evolution and chlorophyll a fluorescence were measured in 1-year-old needles of unfertilized and fertilized trees of Norway spruce (Picea abies (L.) Karst.) during recovery of photosynthesis from winter inhibition in northern Sweden. Measurements were made under laboratory conditions at 20 degrees C. In general, the CO(2)-saturated rate of O(2) evolution was higher in needles of fertilized trees than in needles of unfertilized trees over a wide range of incident photon flux densities. Furthermore, the maximum photochemical efficiency of photosystem (PS) II, as indicated by the ratio of variable to maximum fluorescence (F(V)/F(M)) was higher in needles of fertilized trees than in needles of unfertilized trees. The largest differences in F(V)/F(M) between the two treatments occurred before the main recovery of photosynthesis from winter inhibition in late May. The rate of O(2) evolution was higher in needles of north-facing branches than in needles of south-facing branches in the middle of May. Simultaneous measurements of O(2) exchange and chlorophyll fluorescence indicated that differences in the rate of O(2) evolution between the two treatments were paralleled by differences in the rate of PS II electron transport determined by chlorophyll fluorescence. We suggest that, during recovery of photosynthesis from winter inhibition, the balance between carbon assimilation and PS II electron transport was maintained largely by adjustments in the nonphotochemical dissipation of excitation energy within PS II.

  2. Application of elemental bioimaging using laser ablation ICP-MS in forest pathology: distribution of elements in the bark of Picea sitchensis following wounding.

    Science.gov (United States)

    Siebold, Magdalena; Leidich, Patrick; Bertini, Martina; Deflorio, Giuliana; Feldmann, Jörg; Krupp, Eva M; Halmschlager, Erhard; Woodward, Steve

    2012-04-01

    Element distribution in the bark of two 20-year-old clones of Picea sitchensis following wounding was studied using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Bark was sampled at 0, 3, and 43 days after wounding and analysed using a focused Nd:YAG laser (266 nm). Intensities of (13)C, (25)Mg, (27)Al, (31)P, (32)S, (39)K, (48)Ca, (55)Mn, (57)Fe, (63)Cu and (64)Zn were measured by ICP-MS to study elemental distribution across the bark samples during the wound repair process. A clear accumulation of Mg, P and K at the boundary zone between the lesion and healthy tissue was detected in the wounded samples and was more distinctive at 43 than at 3 days after treatment. This zone of accumulation mapped onto the position of formation of the ligno-suberised boundary zone and differentiation of the wound periderm. These accumulations suggest major roles for Mg, P and K in the non-specific response of Sitka spruce both to wounding, possibly as co-factors to enzymes and energy utilisation. The LA-ICP-MS method developed in this work proved useful to study spatial element distribution across bark samples and has great potential for applications in other areas of plant pathology research.

  3. Application of elemental bioimaging using laser ablation ICP-MS in forest pathology: distribution of elements in the bark of Picea sitchensis following wounding

    International Nuclear Information System (INIS)

    Siebold, Magdalena; Leidich, Patrick; Bertini, Martina; Feldmann, Joerg; Deflorio, Giuliana; Woodward, Steve; Krupp, Eva M.; Halmschlager, Erhard

    2012-01-01

    Element distribution in the bark of two 20-year-old clones of Picea sitchensis following wounding was studied using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Bark was sampled at 0, 3, and 43 days after wounding and analysed using a focused Nd:YAG laser (266 nm). Intensities of 13 C, 25 Mg, 27 Al, 31 P, 32 S, 39 K, 48 Ca, 55 Mn, 57 Fe, 63 Cu and 64 Zn were measured by ICP-MS to study elemental distribution across the bark samples during the wound repair process. A clear accumulation of Mg, P and K at the boundary zone between the lesion and healthy tissue was detected in the wounded samples and was more distinctive at 43 than at 3 days after treatment. This zone of accumulation mapped onto the position of formation of the ligno-suberised boundary zone and differentiation of the wound periderm. These accumulations suggest major roles for Mg, P and K in the non-specific response of Sitka spruce both to wounding, possibly as co-factors to enzymes and energy utilisation. The LA-ICP-MS method developed in this work proved useful to study spatial element distribution across bark samples and has great potential for applications in other areas of plant pathology research. (orig.)

  4. Sequestration of soil nitrogen as tannin-protein complexes may improve the competitive ability of sheep laurel (Kalmia angustifolia) relative to black spruce (Picea mariana).

    Science.gov (United States)

    Joanisse, G D; Bradley, R L; Preston, C M; Bending, G D

    2009-01-01

    The role of litter tannins in controlling soil nitrogen (N) cycling may explain the competitive ability of Kalmia relative to black spruce (Picea mariana), although this has not been demonstrated experimentally. Here, the protein-precipitation capacities of purified tannins and leaf extracts from Kalmia and black spruce were compared. The resistance to degradation of tannin-protein precipitates from both species were compared by monitoring carbon (C) and N dynamics in humus amended with protein, purified tannins or protein-tannin precipitates. The purity of the precipitates was verified using solid-state (13)C nuclear magnetic resonance (NMR) spectra. The ability of mycorrhizal fungi associated with both species to grow on media amended with tannin-protein complexes as the principal N source was also compared. The protein precipitation capacity of Kalmia tannins was superior to those of black spruce. Humus amended with protein increased both mineral and microbial N, whereas humus amended with tannin-protein precipitates increased dissolved organic N. Mycorrhizal fungi associated with Kalmia showed better growth than those associated with black spruce when N was provided as tannin-protein precipitates. These data suggest that Kalmia litter increases the amount of soil N sequestered as tannin-protein complexes, which may improve the competitive ability of Kalmia relative to black spruce by favouring N uptake by mycorrhizas associated with the former.

  5. Application of elemental bioimaging using laser ablation ICP-MS in forest pathology: distribution of elements in the bark of Picea sitchensis following wounding

    Energy Technology Data Exchange (ETDEWEB)

    Siebold, Magdalena [University of Aberdeen, Department of Plant and Soil Science, Institute of Biological and Environmental Sciences, Aberdeen (United Kingdom); University of Natural Resources and Applied Life Sciences Vienna, Department of Forest and Soil Sciences, Institute of Forest Entomology, Forest Pathology and Forest Protection, Vienna (Austria); Georg-August University Goettingen, Division of Plant Pathology and Crop Protection, Department of Crop Sciences, Goettingen (Germany); Leidich, Patrick; Bertini, Martina; Feldmann, Joerg [University of Aberdeen, TESLA, Trace Element Speciation Laboratory, Department of Chemistry, Aberdeen (United Kingdom); Deflorio, Giuliana; Woodward, Steve [University of Aberdeen, Department of Plant and Soil Science, Institute of Biological and Environmental Sciences, Aberdeen (United Kingdom); Krupp, Eva M. [University of Aberdeen, TESLA, Trace Element Speciation Laboratory, Department of Chemistry, Aberdeen (United Kingdom); University of Aberdeen, ACES, Aberdeen Centre of Environmental Sustainability, Aberdeen (United Kingdom); Halmschlager, Erhard [University of Natural Resources and Applied Life Sciences Vienna, Department of Forest and Soil Sciences, Institute of Forest Entomology, Forest Pathology and Forest Protection, Vienna (Austria)

    2012-04-15

    Element distribution in the bark of two 20-year-old clones of Picea sitchensis following wounding was studied using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Bark was sampled at 0, 3, and 43 days after wounding and analysed using a focused Nd:YAG laser (266 nm). Intensities of {sup 13} C, {sup 25}Mg, {sup 27}Al, {sup 31}P, {sup 32}S, {sup 39}K, {sup 48}Ca, {sup 55}Mn, {sup 57}Fe, {sup 63}Cu and {sup 64}Zn were measured by ICP-MS to study elemental distribution across the bark samples during the wound repair process. A clear accumulation of Mg, P and K at the boundary zone between the lesion and healthy tissue was detected in the wounded samples and was more distinctive at 43 than at 3 days after treatment. This zone of accumulation mapped onto the position of formation of the ligno-suberised boundary zone and differentiation of the wound periderm. These accumulations suggest major roles for Mg, P and K in the non-specific response of Sitka spruce both to wounding, possibly as co-factors to enzymes and energy utilisation. The LA-ICP-MS method developed in this work proved useful to study spatial element distribution across bark samples and has great potential for applications in other areas of plant pathology research. (orig.)

  6. Effects of season and urea treatment on infection of stumps of Picea abies by Heterobasidion annosum in stands on former arable land

    Energy Technology Data Exchange (ETDEWEB)

    Brandtberg, P.O. [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Ecology and Environmental Research; Johansson, Martin [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Forest Mycology and Pathology; Seeger, P. [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Statistics

    1996-09-01

    Between 1986 and 1990, a series of thinnings were made in previously unthinned first rotation stands on former arable land located in the southern half of Sweden. The aim was to evaluate the effects of season and urea treatment on the frequency of infection of stumps of Norway spruce (Picea abies (L.) Karst.) by the root-rot fungus Heterobasidion annosum (Fr.) Bref. Untreated stumps, resulting from 60 thinnings (22-100 stumps each, altogether ca 3000 stumps) made at different times of year, were investigated 3-24 months after cutting to determine whether they were infected with H. annosum. On average only 2% of the stumps from thinnings made in November-February were infected, whereas the incidence of infection among stumps thinned in June-July was 34%. Two methods of treating stumps with urea to prevent stump infection by H. annosum after thinning were evaluated in terms of effectiveness. The freshly cut stumps were treated with a 20% urea solution, transformed to a gel by adding 0.2% carboxymethyl cellulose, or with a 30% urea solution. On average, the reduction in infection rate obtained was 62% with the first method and 85% with the latter. In a separate study involving a concentration series of urea, there was a considerable drop in protection efficiency, from 89% to 58%, when the concentration was decreased from 30% to 15%. 38 refs, 3 figs, 1 tab

  7. Contamination of environment in the road surroudings - impact of road salting on Norway spruce (Picea abies) and Scots pine (Pinus sylvestris)

    Science.gov (United States)

    Hegrová, Jitka; Steiner, Oliver; Goessler, Walter; Tanda, Stefan; Anděl, Petr

    2017-09-01

    A comprehensive overview of the influence of transport on the environment is presented in this study. The complex analysis of soil and needle samples provides an extensive set of data, which presents elemental contamination of the environment near roads. Traffic pollution (including winter road treatment) has a significant negative influence on our environment. Besides sodium and chlorine from winter maintenance many other elements are emitted into the environment. Three possible sources of contamination are assumed for environmental contamination evaluation: car emission, winter maintenance and abrasion from breaks and clutches. The chemical analysis focused on the description of samples from inorganic point of view. The influence of the contamination potential on the sodium and chlorine content in the samples of 1st year-old and 2nd year-old needles of Norway spruce (Picea abies) and Scots pine (Pinus sylvestris) is discussed. Additional soil samples were taken from each sampling site and analyzed to get insight in the sodium and chlorine distribution. Statistical evaluation was used for interpretation of complex interaction patterns between element concentrations in different aged needles based on localities character including distance from the road and element concentration in soils. This species of needles were chosen because of its heightened sensitivity towards salinization. The study was conducted in different parts of the Czech Republic. The resulting database is a source of valuable information about the influence of transport on the environment.

  8. Dynamics at the treeline: differential responses of Picea mariana and Larix laricina to climate change in eastern subarctic Québec

    International Nuclear Information System (INIS)

    Dufour-Tremblay, Geneviève; Boudreau, Stéphane; Lévesque, Esther

    2012-01-01

    Treelines are known to be temperature-sensitive ecotones, and therefore could rapidly expand their range limits in response to climate warming. Observations of lack of range expansion, however, indicate that ecological constraints partly control the treeline ecotones. The main objectives of this study were to evaluate Picea mariana and Larix laricina recruitment and growth at and above the altitudinal treeline of Kangiqsualujjuaq (Nunavik), where warmer temperatures since the 1990s have already triggered shrub expansion. We mapped, harvested, dated and measured tree individuals along two altitudinal gradients from the forested stands below the treeline to hilltops. Since the 1990s, a pulse of L. laricina seedling establishment has occurred at and above the treeline. Dendrochronological analysis revealed that L. laricina underwent a rapid vertical growth and radial growth that accelerated from the 1990s. No recruitment was observed for P. mariana in response to the regional warming, suggesting a regeneration failure of this species. Our results indicated that the L. laricina colonization below and above the treeline in recent decades in response to the regional warming should modify the landscape physiognomy of the study area in the near future. (letter)

  9. The effects of wind and temperature on cuticular transpiration of Picea abies and Pinus cembra and their significance in dessication damage at the alpine treeline.

    Science.gov (United States)

    Baig, M N; Tranquillini, W

    1980-01-01

    The importance of high winter winds and plant temperatures as causes of winter desiccation damage at the alpine treeline were studied in the Austrian Alps. Samples of 1- and 2-year twigs of Picea abies and Pinus cembra were collected from the valley bottom (1,000 m a.s.l.), forestline (1,940 m a.s.l.), kampfzone (2.090 m a.s.l.), wind-protected treeline (2,140 m a.s.l.), and wind-exposed treeline (2,140 m a.s.l.). Cuticular transpiration was measured at three different levels of wind speed (4, 10, and 15 ms -1 ) and temperature (15°, 20°, and 25° C). At elevated wind speeds slight increases in water loss were observed, whereas at higher temperatures much greater increases occurred. Studies on winter water relations show a significant decline in the actual moisture content and osmotic potentials of twigs, especially in the kampfzone and at treeline. The roles of high winds and temperatures in depleting the winter water economy and causing desiccation damage in the alpine treeline environment are discussed.

  10. Topography- and Species-Dependent Climatic Responses in Radial Growth of Picea meyeri and Larix principis-rupprechtii in the Luyashan Mountains of North-Central China

    Directory of Open Access Journals (Sweden)

    Wentao Zhang

    2015-01-01

    Full Text Available Dendroecological techniques were used to examine the relationships between topographic aspects, climate factors and radial growth of Picea meyeri and Larix principis-rupprechtii in Luyashan Mountains, North-Central China. Four sites were selected at timberline and totally 67 trees and 134 cores were collected. Pearson correlation and regression surface analysis were conducted to reveal the growth-climate relationships. The results indicated that the two species both showed significant negative correlations with temperature during preceding November on the two topographic aspects. On both slope aspects, growth of P. meyeri exhibited significant negative correlations with precipitation in current June, whereas growth of L. principis-rupprechtii showed significant negative correlations with precipitation in preceding September. On north-facing slope, tree growth was limited by low temperature in early growing season, which not shown on south-facing slope. If climate warming continues, L. principis-rupprechtii may be more favored and a reverse between relationships with temperature and precipitation maybe occur in growth of trees. Treeline position on the north-facing slope may possess a greater potential for elevation shifting than the south-facing slope. Our results supply useful information for discussing the potential effect of future climate on the forest growth in North-Central China.

  11. Impact of climate change, seedling type and provenance on the risk of damage to Norway spruce (Picea abies (L.) Karst.) seedlings in Sweden due to early summer frosts

    Energy Technology Data Exchange (ETDEWEB)

    Langvall, Ola (Swedish Univ. of Agricultural Sciences, Unit for Field-based Forest Research, Asa Forest Research Station, Lammhult (Sweden))

    2011-04-15

    A model including site-specific microclimate-affecting properties of a forest regeneration area together with seedling characteristics was used to evaluate the accumulated risk of frost damage to Norway spruce (Picea abies (L.) Karst.) seedlings. Climate change in Sweden was simulated on the basis of the regional climate model RCA3. The daily average temperature, the driving factor for bud burst in the model, was adjusted using the difference between the mean of the climate model data for the years 1961-1990 and 2036-2065. The model was run for a highly frost prone, clear-cut site in which bare-rooted Norway spruce seedlings of mid-Swedish provenance were planted. Alternate runs were conducted with data for containerized seedlings and seedlings of Belarusian origin. The study showed that bud burst will occur at earlier dates throughout Sweden in the period 2036-2065 if the climate changes according to either of the climate scenarios examined, compared to the reference period 1961-1990. Furthermore, the risk of damage to Norway spruce seedlings as a result of frost events during summer will increase in southern Sweden and be unaffected or decrease in northern Sweden. The risk of frost damage was exacerbated in containerized seedlings, while the risk was lower for the seedlings of Belarusian provenance when compared with bare-rooted seedlings or seedlings of mid-Swedish origin

  12. Methylated Fatty Acids from Heartwood and Bark of Pinus sylvestris, Abies alba, Picea abies, and Larix decidua: Effect of Strong Acid Treatment

    Directory of Open Access Journals (Sweden)

    Mohamed Zidan Mohamed Salem

    2015-09-01

    Full Text Available Methylated fatty acid (FA compounds in the heartwood and bark of some softwood species, specifically Pinus sylvestris, Abies alba, Picea abies, and Larix decidua, grown in the Czech Republic were evaluated. Strong H2SO4 was used for methylation of the lipids. The highest content of lipid was found in P. abies bark (40.132 mg/g o.d. sample, and the lowest content was in A. alba wood (11.027 mg/g o.d. sample. The highest concentration of FAs was observed in L. decidua bark. The highest percentages of FAs in wood of P. sylvestris were arachidic acid and oleic acid. In bark, the highest percentages of FAs were stearic acid, palmitic acid, and oleic acid. The FAs with the highest concentrations in A. alba wood were arachidic acid, palmitic acid, pentadecanoic acid, and margarinic, and those in bark were behenic acid, lignoceric acid, and arachidic acid. P. abies wood FAs showed arachidic acid, palmitic acid, and margarinic acid, and the bark contained lignoceric acid and arachidic acid. The FAs of L. decidua wood were arachidic acid, palmitic acid, and stearic acid, and in bark they were pentacosylic acid, docosahexaenoic acid (DHA, lignoceric acid, arachidic acid, and behenic acid. The lack of typically dominant unsaturated fatty acids (e.g. 18:1, 18:2, compared to literature values were attributed to the application of strong acid for the hydrolysis.

  13. Atmospheric and geogenic CO2 within the crown and root of spruce (Picea abies L. Karst.) growing in a mofette area

    Science.gov (United States)

    Vodnik, D.; Thomalla, A.; Ferlan, M.; Levanič, T.; Eler, K.; Ogrinc, N.; Wittmann, C.; Pfanz, H.

    2018-06-01

    Mofettes are often investigated in ecology, either as extreme sites, natural analogues to future conditions under climate change, or model ecosystems for environmental impact assessments of carbon capture and storage systems. Much of this research, however, inadequately addresses the complexity of the gas environment at these sites, mainly focusing on aboveground CO2-enrichment. In the current research, the gaseous environment of Norway spruce (Picea abies (L) Karst.) trees growing at the Stavešinske slepice mofette (NE Slovenia) were studied by measuring both soil ([CO2]soil) and atmospheric CO2 concentrations ([CO2]air). Within the studied site (800 m2), soil CO2 enrichment was spatially heterogeneous; about 25% of the area was characterized by very high [CO2]soil (>40%) and hypoxic conditions. Aboveground gas measurements along vertical profiles not only revealed substantially elevated [CO2]air close to the ground (height up to 1.5 m), but also in the upper heights (20-25 m; crown layer). On the basis δ13C of CO2, it was shown that elevated CO2 relates to a geogenic source. Trees grown in high [CO2]soil were characterized by decreased radial growth; the δ13C of their wood was less negative than in trees growing in normal soil. Unfavorable gaseous soil conditions should generally be accepted as being by far the most important factor affecting (i.e. disturbing) the growth of mofette trees.

  14. Metabolite changes in conifer buds and needles during forced bud break in Norway spruce (Picea abies and European silver fir (Abies alba

    Directory of Open Access Journals (Sweden)

    Priyanka eDhuli

    2014-12-01

    Full Text Available Environmental changes such as early spring and warm spells induce bud burst and photosynthetic processes in cold-acclimated coniferous trees and consequently, cellular metabolism in overwintering needles and buds. The purpose of the study was to examine metabolism in conifers under forced deacclimation (artificially induced spring by exposing shoots of Picea abies (boreal species and Abies alba (temperate species to a greenhouse environment (22°C, 16/8 h D/N cycle over a nine week period. Each week, we scored bud opening and collected samples for GC/MS–based metabolite profiling. We detected a total of 169 assigned metabolites and 80 identified metabolites, comprising compounds such as mono- and disaccharides, Krebs cycle acids, amino acids, polyols, phenolics and phosphorylated structures. Untargeted multivariate statistical analysis based on PCA and cluster analysis segregated samples by species, tissue type, and stage of tissue deacclimations. Similar patterns of metabolic regulation in both species were observed in buds (amino acids, Krebs cycle acids and needles (hexoses, pentoses, and Krebs cycle acids. Based on correlation of bud opening score with compound levels, distinct metabolites could be associated with bud and shoot development, including amino acids, sugars and acids with known osmolyte function, and secondary metabolites. This study has shed light on how elevated temperature affects metabolism in buds and needles of conifer species during the deacclimation phase, and contributes to the discussion about how phenological characters in conifers may respond to future global warming.

  15. Morphological and ecological variation of Gremmeniella abietina var. abietina in Pinus sylvestris, Pinus contorta and Picea abies sapling stands in northern Finland and the Kola Peninsula

    Energy Technology Data Exchange (ETDEWEB)

    Kaitera, Juha; Seitamaeki, Leena; Jalkanen, Risto [Finnish Forest Research Inst., Rovaniemi (Finland). Rovaniemi Research Station

    2000-07-01

    The morphological and ecological variation of two types of Gremmeniella abietina var. abietina causing scleroderris canker on conifers was investigated in Pinus spp. and Picea sp. sapling stands in northern Finland and the Kola Peninsula. Small-tree type (STT or B type) of G. abietina was detected alone in 13 Scots pine, three lodgepole pine and two Norway spruce sapling stands out of 26 stands investigated, both STT and large-tree type (LTT or A type) were observed in six Scots pine stands, and LTT was detected alone in two Scots pine stands. For the first time, G. abietina was found to injure Norway spruce saplings in a respective plantation in northern Fennoscandia. STT isolates produced statistically significantly more conidia in vitro than LTT isolates. Morphological variation in conidia septation revealed that STT produced conidia with more than five septa more frequently than did LTT. There was a greater range in variation in septation in STT than in LTT, with overlapping between the types. Isolates of both types were equally associated with cankers, coloured wood, pycnidia or apothecia in the infected saplings.

  16. Differences in the photosynthetic UV-B response between European beech (Fagus sylvatica L.) and Norway spruce (Picea abies (L.) Karst.) saplings

    International Nuclear Information System (INIS)

    Šprtová, M.; Marek, M.V.; Urban, O.; Kalina, J.; Špunda, V.

    2008-01-01

    Cloned saplings of Norway spruce (Picea abies (L) Karst.) and beech (Fagus sylvatica L.) (7 years old) were exposed to enhanced UV-B irradiation (+25%) continuously over three growing seasons (1999-2001). Selected parameters of variable chlorophyll alpha fluorescence and pigment composition were analysed in the late summer of the third growing season to evaluate the influence of long-term elevated UV-B irradiation on broadleaf and conifer tree species. To obtain information on the xanthophyll cycle, the de-epoxidation state (DEPS) was calculated. These tree species responded differentially to the long-term effects of enhanced UV-B radiation, Norway spruce was more sensitive compared to the European beech. The results show that in Norway spruce long-term exposure to enhanced UV-B radiation under field conditions caused negative changes at the level of primary photosynthetic reactions. Contrary to the beech, this had higher degree of UV-B protective responses. UV-B radiation is not effective stressor to its primary photosynthetic reaction

  17. Development of a Dispersal Model for Balsam Woolly Adelgid, Adelges piceae Ratzeburg (Hemiptera: Adelgidae, to Facilitate Landscape-Level Management Planning

    Directory of Open Access Journals (Sweden)

    L. W. Lass

    2014-01-01

    Full Text Available The balsam woolly adelgid (Adelges piceae Ratzeburg attacks subalpine fir (Abies lasiocarpa (Hook. Nutt. in eastern Washington, Oregon, and northern Idaho. Historical balsam woolly adelgid distributions present an opportunity to understand climatic factors that influence the species’ distribution at a landscape scale. The distribution data allows for creation of predictive models that detail the likelihood of occurrence and associated geographic data allow modeling of species dispersal. Predictive variables linked to the distribution of the hosts and to abiotic environmental conditions were utilized to create a spatial probability model of occurrence. Balsam woolly adelgid predominantly disperses by wind, and hence, both wind speed and wind direction were used to create a dispersal probability model. Results from wind dispersal modeling suggested that two-thirds of the new infestations were due to July and August wind direction and speed. Average July winds ranged from 0.5 to 3.27 m/s, flowing south westerly, and August winds ranged from 0.43 to 1.55 m/s, flowing north easterly. Land managers can use the results of the predictive model to better understand where current infestations are likely to expand. Prediction of where the balsam woolly adelgid might move allows managers to adjust actions to respond to future insect movement and establishment.

  18. Isolation of BAC Clones Containing Conserved Genes from Libraries of Three Distantly Related Moths: A Useful Resource for Comparative Genomics of Lepidoptera

    Directory of Open Access Journals (Sweden)

    Yuji Yasukochi

    2011-01-01

    Full Text Available Lepidoptera, butterflies and moths, is the second largest animal order and includes numerous agricultural pests. To facilitate comparative genomics in Lepidoptera, we isolated BAC clones containing conserved and putative single-copy genes from libraries of three pests, Heliothis virescens, Ostrinia nubilalis, and Plutella xylostella, harboring the haploid chromosome number, =31, which are not closely related with each other or with the silkworm, Bombyx mori, (=28, the sequenced model lepidopteran. A total of 108–184 clones representing 101–182 conserved genes were isolated for each species. For 79 genes, clones were isolated from more than two species, which will be useful as common markers for analysis using fluorescence in situ hybridization (FISH, as well as for comparison of genome sequence among multiple species. The PCR-based clone isolation method presented here is applicable to species which lack a sequenced genome but have a significant collection of cDNA or EST sequences.

  19. BacMam Virus Transduced Cardiomyoblasts Can Be Used for Myocardial Transplantation Using AP-PEG-A Microcapsules: Molecular Cloning, Preparation, and In Vitro Analysis

    Directory of Open Access Journals (Sweden)

    Arghya Paul

    2010-01-01

    Full Text Available The potential of genetically modified cardiomyoblasts in treating damaged myocardium is well known. However, efficient delivery of these cells is of major concern during treatment. The limiting factors are the massive cell death that occurs soon after their intramyocardial transplantation into the beating heart. To address these problems, we generated recombinant baculoviruses (BacMam viruses which efficiently transduced cardiomyoblast cells under optimized conditions. These genetically modified cells were then protected in a new polymeric microcapsule using poly-ethylene-glycol (PEG, alginate, and poly-L-lysine (PLL polymers for efficient delivery. Results showed that microcapsules maintain cell viability and support cell proliferation for at least 30 days. The capsules exhibit strong immunoprotective potential and have high mechanical and osmotic stability with more than 70% intact capsules. The encased transduced cells showed a rapid transgene expression inside the capsule for at least 15 days. However, preclinical studies are needed to further explore its long-term functional benefits.

  20. BacMam virus transduced cardiomyoblasts can be used for myocardial transplantation using AP-PEG-A microcapsules: molecular cloning, preparation, and in vitro analysis.

    Science.gov (United States)

    Paul, Arghya; Khan, Afshan Afsar; Shum-Tim, Dominique; Prakash, Satya

    2010-01-01

    The potential of genetically modified cardiomyoblasts in treating damaged myocardium is well known. However, efficient delivery of these cells is of major concern during treatment. The limiting factors are the massive cell death that occurs soon after their intramyocardial transplantation into the beating heart. To address these problems, we generated recombinant baculoviruses (BacMam viruses) which efficiently transduced cardiomyoblast cells under optimized conditions. These genetically modified cells were then protected in a new polymeric microcapsule using poly-ethylene-glycol (PEG), alginate, and poly-L-lysine (PLL) polymers for efficient delivery. Results showed that microcapsules maintain cell viability and support cell proliferation for at least 30 days. The capsules exhibit strong immunoprotective potential and have high mechanical and osmotic stability with more than 70% intact capsules. The encased transduced cells showed a rapid transgene expression inside the capsule for at least 15 days. However, preclinical studies are needed to further explore its long-term functional benefits.

  1. FISH analysis of the W chromosome in Bombyx mandarina and several other species of Lepidoptera by means of B. mori W-BAC probes

    Czech Academy of Sciences Publication Activity Database

    Yoshido, A.; Yasukochi, Y.; Marec, František; Abe, H.; Sahara, K.

    2007-01-01

    Roč. 76, č. 1 (2007), s. 1-7 ISSN 1346-8073 R&D Projects: GA ČR GA206/06/1860; GA AV ČR IAA6007307 Grant - others:Japan Society for the Promotion of Science(JP) 15380227; Japan Society for the Promotion of Science(JP) 18380037; Japan Society for the Promotion of Science(JP) 15658020; Ministry of Agriculture, Forestry and Fisheries in Japan(JP) 2102(Y.Y.); Ministry of Agriculture, Forestry and Fisheries in Japan(JP) 2013(K.S.) Institutional research plan: CEZ:AV0Z50070508 Keywords : BAC-FISH * fluorescence in situ hybridization * GISH Subject RIV: EB - Genetics ; Molecular Biology

  2. Inducible targeting of CNS astrocytes in Aldh1l1-CreERT2 BAC transgenic mice [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Jan Winchenbach

    2016-12-01

    Full Text Available Background: Studying astrocytes in higher brain functions has been hampered by the lack of genetic tools for the efficient expression of inducible Cre recombinase throughout the CNS, including the neocortex. Methods: Therefore, we generated BAC transgenic mice, in which CreERT2 is expressed under control of the Aldh1l1 regulatory region. Results: When crossbred to Cre reporter mice, adult Aldh1l1-CreERT2 mice show efficient gene targeting in astrocytes. No such Cre-mediated recombination was detectable in CNS neurons, oligodendrocytes, and microglia. As expected, Aldh1l1-CreERT2 expression was evident in several peripheral organs, including liver and kidney. Conclusions: Taken together, Aldh1l1-CreERT2 mice are a useful tool for studying astrocytes in neurovascular coupling, brain metabolism, synaptic plasticity and other aspects of neuron-glia interactions.

  3. Oligonucleotide arrays vs. metaphase-comparative genomic hybridisation and BAC arrays for single-cell analysis: first applications to preimplantation genetic diagnosis for Robertsonian translocation carriers.

    Science.gov (United States)

    Ramos, Laia; del Rey, Javier; Daina, Gemma; García-Aragonés, Manel; Armengol, Lluís; Fernandez-Encinas, Alba; Parriego, Mònica; Boada, Montserrat; Martinez-Passarell, Olga; Martorell, Maria Rosa; Casagran, Oriol; Benet, Jordi; Navarro, Joaquima

    2014-01-01

    Comprehensive chromosome analysis techniques such as metaphase-Comparative Genomic Hybridisation (CGH) and array-CGH are available for single-cell analysis. However, while metaphase-CGH and BAC array-CGH have been widely used for Preimplantation Genetic Diagnosis, oligonucleotide array-CGH has not been used in an extensive way. A comparison between oligonucleotide array-CGH and metaphase-CGH has been performed analysing 15 single fibroblasts from aneuploid cell-lines and 18 single blastomeres from human cleavage-stage embryos. Afterwards, oligonucleotide array-CGH and BAC array-CGH were also compared analysing 16 single blastomeres from human cleavage-stage embryos. All three comprehensive analysis techniques provided broadly similar cytogenetic profiles; however, non-identical profiles appeared when extensive aneuploidies were present in a cell. Both array techniques provided an optimised analysis procedure and a higher resolution than metaphase-CGH. Moreover, oligonucleotide array-CGH was able to define extra segmental imbalances in 14.7% of the blastomeres and it better determined the specific unbalanced chromosome regions due to a higher resolution of the technique (≈ 20 kb). Applicability of oligonucleotide array-CGH for Preimplantation Genetic Diagnosis has been demonstrated in two cases of Robertsonian translocation carriers 45,XY,der(13;14)(q10;q10). Transfer of euploid embryos was performed in both cases and pregnancy was achieved by one of the couples. This is the first time that an oligonucleotide array-CGH approach has been successfully applied to Preimplantation Genetic Diagnosis for balanced chromosome rearrangement carriers.

  4. Oligonucleotide arrays vs. metaphase-comparative genomic hybridisation and BAC arrays for single-cell analysis: first applications to preimplantation genetic diagnosis for Robertsonian translocation carriers.

    Directory of Open Access Journals (Sweden)

    Laia Ramos

    Full Text Available Comprehensive chromosome analysis techniques such as metaphase-Comparative Genomic Hybridisation (CGH and array-CGH are available for single-cell analysis. However, while metaphase-CGH and BAC array-CGH have been widely used for Preimplantation Genetic Diagnosis, oligonucleotide array-CGH has not been used in an extensive way. A comparison between oligonucleotide array-CGH and metaphase-CGH has been performed analysing 15 single fibroblasts from aneuploid cell-lines and 18 single blastomeres from human cleavage-stage embryos. Afterwards, oligonucleotide array-CGH and BAC array-CGH were also compared analysing 16 single blastomeres from human cleavage-stage embryos. All three comprehensive analysis techniques provided broadly similar cytogenetic profiles; however, non-identical profiles appeared when extensive aneuploidies were present in a cell. Both array techniques provided an optimised analysis procedure and a higher resolution than metaphase-CGH. Moreover, oligonucleotide array-CGH was able to define extra segmental imbalances in 14.7% of the blastomeres and it better determined the specific unbalanced chromosome regions due to a higher resolution of the technique (≈ 20 kb. Applicability of oligonucleotide array-CGH for Preimplantation Genetic Diagnosis has been demonstrated in two cases of Robertsonian translocation carriers 45,XY,der(13;14(q10;q10. Transfer of euploid embryos was performed in both cases and pregnancy was achieved by one of the couples. This is the first time that an oligonucleotide array-CGH approach has been successfully applied to Preimplantation Genetic Diagnosis for balanced chromosome rearrangement carriers.

  5. Oligonucleotide Arrays vs. Metaphase-Comparative Genomic Hybridisation and BAC Arrays for Single-Cell Analysis: First Applications to Preimplantation Genetic Diagnosis for Robertsonian Translocation Carriers

    Science.gov (United States)

    Ramos, Laia; del Rey, Javier; Daina, Gemma; García-Aragonés, Manel; Armengol, Lluís; Fernandez-Encinas, Alba; Parriego, Mònica; Boada, Montserrat; Martinez-Passarell, Olga; Martorell, Maria Rosa; Casagran, Oriol; Benet, Jordi; Navarro, Joaquima

    2014-01-01

    Comprehensive chromosome analysis techniques such as metaphase-Comparative Genomic Hybridisation (CGH) and array-CGH are available for single-cell analysis. However, while metaphase-CGH and BAC array-CGH have been widely used for Preimplantation Genetic Diagnosis, oligonucleotide array-CGH has not been used in an extensive way. A comparison between oligonucleotide array-CGH and metaphase-CGH has been performed analysing 15 single fibroblasts from aneuploid cell-lines and 18 single blastomeres from human cleavage-stage embryos. Afterwards, oligonucleotide array-CGH and BAC array-CGH were also compared analysing 16 single blastomeres from human cleavage-stage embryos. All three comprehensive analysis techniques provided broadly similar cytogenetic profiles; however, non-identical profiles appeared when extensive aneuploidies were present in a cell. Both array techniques provided an optimised analysis procedure and a higher resolution than metaphase-CGH. Moreover, oligonucleotide array-CGH was able to define extra segmental imbalances in 14.7% of the blastomeres and it better determined the specific unbalanced chromosome regions due to a higher resolution of the technique (≈20 kb). Applicability of oligonucleotide array-CGH for Preimplantation Genetic Diagnosis has been demonstrated in two cases of Robertsonian translocation carriers 45,XY,der(13;14)(q10;q10). Transfer of euploid embryos was performed in both cases and pregnancy was achieved by one of the couples. This is the first time that an oligonucleotide array-CGH approach has been successfully applied to Preimplantation Genetic Diagnosis for balanced chromosome rearrangement carriers. PMID:25415307

  6. Changes in the Metabolome of Picea balfouriana Embryogenic Tissues That Were Linked to Different Levels of 6-BAP by Gas Chromatography-Mass Spectrometry Approach.

    Directory of Open Access Journals (Sweden)

    Q F Li

    Full Text Available Embryogenic cultures of Picea balfouriana, which is an important commercial species for reforestation in Southern China, easily lose their embryogenic ability during long-term culture. Embryogenic tissue that proliferated at lower concentrations (3.6 μM and 2.5 μM of 6-benzylaminopurine (6-BAP were more productive, and generated 113 ± 6 and 89 ± 3 mature embryos per 100 mg embryogenic tissue, respectively. A metabolomic approach was used to study the changes in metabolites linked to embryogenic competence related to three different 6-BAP concentrations (2.5 μM, 3.6 μM, and 5 μM. A total of 309 compounds were obtained, among which 123 metabolites mapped to Kyoto Encyclopedia of Genes and genomes (KEGG pathways. The levels of 35 metabolites were significantly differentially regulated among the three 6-BAP treatments, and 32 metabolites differed between the 2.5 μM and 5 μM treatments. A total of 17 metabolites appeared only once among the three comparisons. The combination of a score plot and a loading plot showed that in the samples with higher embryogenic ability (3.6 μM and 2.5 μM, up-regulated metabolites were mostly amino acids and down-regulated metabolites were mostly primary carbohydrates (especially sugars. These results suggested that 6-BAP may influence embryogenic competence by nitrogen metabolism, which could cause an increase in amino acid levels and higher amounts of aspartate, isoleucine, and leucine in tissues with higher embryogenic ability. Furthermore, we speculated that 6-BAP may affect the amount of tryptophan in tissues, which would change the indole-3-acetic acid levels and influence the embryogenic ability.

  7. A Picea crassifolia Tree-Ring Width-Based Temperature Reconstruction for the Mt. Dongda Region, Northwest China, and Its Relationship to Large-Scale Climate Forcing.

    Directory of Open Access Journals (Sweden)

    Yu Liu

    Full Text Available The historical May-October mean temperature since 1831 was reconstructed based on tree-ring width of Qinghai spruce (Picea crassifolia Kom. collected on Mt. Dongda, North of the Hexi Corridor in Northwest China. The regression model explained 46.6% of the variance of the instrumentally observed temperature. The cold periods in the reconstruction were 1831-1889, 1894-1901, 1908-1934 and 1950-1952, and the warm periods were 1890-1893, 1902-1907, 1935-1949 and 1953-2011. During the instrumental period (1951-2011, an obvious warming trend appeared in the last twenty years. The reconstruction displayed similar patterns to a temperature reconstruction from the east-central Tibetan Plateau at the inter-decadal timescale, indicating that the temperature reconstruction in this study was a reliable proxy for Northwest China. It was also found that the reconstruction series had good consistency with the Northern Hemisphere temperature at a decadal timescale. Multi-taper method spectral analysis detected some low- and high-frequency cycles (2.3-2.4-year, 2.8-year, 3.4-3.6-year, 5.0-year, 9.9-year and 27.0-year. Combining these cycles, the relationship of the low-frequency change with the Pacific Decadal Oscillation (PDO, North Atlantic Oscillation (NAO and Southern Oscillation (SO suggested that the reconstructed temperature variations may be related to large-scale atmospheric-oceanic variations. Major volcanic eruptions were partly reflected in the reconstructed temperatures after high-pass filtering; these events promoted anomalous cooling in this region. The results of this study not only provide new information for assessing the long-term temperature changes in the Hexi Corridor of Northwest China, but also further demonstrate the effects of large-scale atmospheric-oceanic circulation on climate change in Northwest China.

  8. Effect of Forest Management of Picea abies and Fagus sylvatica with Different Types of Felling on Carbon and Economic Balances in the Czech Republic

    Science.gov (United States)

    Plch, Radek; Pulkrab, Karel; Bukáček, Jan; Sloup, Roman; Cudlín, Pavel

    2016-10-01

    The selection of the most sustainable forest management under given site conditions needs suitable criteria and indicators. For this purpose, carbon and economic balance assessment, completed with environmental impact computation using the Life Cycle Assessment (LCA) were used. The aim of this study was to compare forestry operations and wood production of selected forest stands with different i) tree species composition (Norway spruce - Picea abies and European beech - Fagus sylvatica) and ii) type of felling (chainsaw and harvester). Carbon and economic balance methods consist in the comparison of quantified inputs (fossil fuels, electricity, used machinery, fertilizers, etc., converted into emission units of carbon in Mg of C- CO2-eq. or EUR) with quantified outputs (biomass production in Mg of carbon or EUR). In this contribution, similar forest stands (“forest site complexes”) in the 4th forest vegetation zone (in the Czech Republic approximately 400-700 m above sea-level) were selected. Forestry operations were divided into 5 main stages: i) seedling production, ii) stand establishment and pruning, iii) thinning and final cutting, iv) skidding, and v) secondary timber transport and modelled for one rotation period of timber production (ca. 100 years). The differences between Norway spruce and European beech forest stands in the carbon efficiency were relatively small while higher differences were achieved in the economic efficiency (forest stands with Norway spruce had a higher economic efficiency). Concerning the comparison of different types of felling in Norway spruce forest stands, the harvester use proved to induce significantly higher environmental impacts (emission of carbon) and lower economic costs. The comparison of forestry operation stages showed that the main part of carbon emissions, originating from fuel production and combustion, is connected with a thinning and final cutting, skidding and secondary timber transport in relations to

  9. Assessing the impacts of climate change and nitrogen deposition on Norway spruce (Picea abies L. Karst) growth in Austria with BIOME-BGC.

    Science.gov (United States)

    Eastaugh, Chris S; Pötzelsberger, Elisabeth; Hasenauer, Hubert

    2011-03-01

    The aim of this paper is to determine whether a detectable impact of climate change is apparent in Austrian forests. In regions of complex terrain such as most of Austria, climatic trends over the past 50 years show marked geographic variability. As climate is one of the key drivers of forest growth, a comparison of growth characteristics between regions with different trends in temperature and precipitation can give insights into the impact of climatic change on forests. This study uses data from several hundred climate recording stations, interpolated to measurement sites of the Austrian National Forest Inventory (NFI). Austria as a whole shows a warming trend over the past 50 years and little overall change in precipitation. The warming trends, however, vary considerably across certain regions and regional precipitation trends vary widely in both directions, which cancel out on the national scale These differences allow the delineation of 'climatic change zones' with internally consistent climatic trends that differ from other zones. This study applies the species-specific adaptation of the biogeochemical model BIOME-BGC to Norway spruce (Picea abies (L.) Karst) across a range of Austrian climatic change zones, using input data from a number of national databases. The relative influence of extant climate change on forest growth is quantified, and compared with the far greater impact of non-climatic factors. At the national scale, climate change is found to have negligible effect on Norway spruce productivity, due in part to opposing effects at the regional level. The magnitudes of the modeled non-climatic influences on aboveground woody biomass increment increases are consistent with previously reported values of 20-40 kg of added stem carbon sequestration per kilogram of additional nitrogen deposition, while climate responses are of a magnitude difficult to detect in NFI data.

  10. Online investigation of respiratory quotients in Pinus sylvestris and Picea abies during drought and shading by means of cavity-enhanced Raman multi-gas spectrometry.

    Science.gov (United States)

    Hanf, Stefan; Fischer, Sarah; Hartmann, Henrik; Keiner, Robert; Trumbore, Susan; Popp, Jürgen; Frosch, Torsten

    2015-07-07

    Photosynthesis and respiration are major components of the plant carbon balance. During stress, like drought, carbohydrate supply from photosynthesis is reduced and the Krebs cycle respiration must be fueled with other stored carbon compounds. However, the dynamics of storage use are still unknown. The respiratory quotient (RQ, CO2 released per O2 consumed during respiration) is an excellent indicator of the nature of the respiration substrate. In plant science, however, online RQ measurements have been challenging or even impossible so far due to very small gas exchange fluxes during respiration. Here we apply cavity-enhanced multi-gas Raman spectrometry (CERS) for online in situ RQ measurements in drought-tolerant pine (Pinus sylvestris [L.]) and drought-intolerant spruce (Picea abies [L. H. Karst]). Two different treatments, drought and shading, were applied to reduce photosynthesis and force dependency on stored substrates. Changes in respiration rates and RQ values were continuously monitored over periods of several days with low levels of variance. The results show that both species switched from COH-dominated respiration (RQ = 1.0) to a mixture of substrates during shading (RQ = 0.77-0.81), while during drought only pine did so (RQ = 0.75). The gas phase measurements were complemented by concentration measurements of non-structural carbohydrates and lipids. These first results suggest a physiological explanation for greater drought tolerance in pine. CERS was proven as powerful technique for non-consumptive and precise real-time monitoring of respiration rates and respirational quotients for the investigation of plant metabolism under drought stress conditions that are predicted to increase with future climate change.

  11. Predicting Effects of Climate Change on Habitat Suitability of Red Spruce (Picea rubens Sarg. in the Southern Appalachian Mountains of the USA: Understanding Complex Systems Mechanisms through Modeling

    Directory of Open Access Journals (Sweden)

    Kyung Ah Koo

    2015-04-01

    Full Text Available Alpine, subalpine and boreal tree species, of low genetic diversity and adapted to low optimal temperatures, are vulnerable to the warming effects of global climate change. The accurate prediction of these species’ distributions in response to climate change is critical for effective planning and management. The goal of this research is to predict climate change effects on the distribution of red spruce (Picea rubens Sarg. in the Great Smoky Mountains National Park (GSMNP, eastern USA. Climate change is, however, conflated with other environmental factors, making its assessment a complex systems problem in which indirect effects are significant in causality. Predictions were made by linking a tree growth simulation model, red spruce growth model (ARIM.SIM, to a GIS spatial model, red spruce habitat model (ARIM.HAB. ARIM.SIM quantifies direct and indirect interactions between red spruce and its growth factors, revealing the latter to be dominant. ARIM.HAB spatially distributes the ARIM.SIM simulations under the assumption that greater growth reflects higher probabilities of presence. ARIM.HAB predicts the future habitat suitability of red spruce based on growth predictions of ARIM.SIM under climate change and three air pollution scenarios: 10% increase, no change and 10% decrease. Results show that suitable habitats shrink most when air pollution increases. Higher temperatures cause losses of most low-elevation habitats. Increased precipitation and air pollution produce acid rain, which causes loss of both low- and high-elevation habitats. The general prediction is that climate change will cause contraction of red spruce habitats at both lower and higher elevations in GSMNP, and the effects will be exacerbated by increased air pollution. These predictions provide valuable information for understanding potential impacts of global climate change on the spatiotemporal distribution of red spruce habitats in GSMNP.

  12. Fine root biomass, necromass and chemistry during seven years of elevated aluminium concentrations in the soil solution of a middle-aged Picea abies stand.

    Science.gov (United States)

    Eldhuset, Toril D; Lange, Holger; de Wit, Helene A

    2006-10-01

    Toxic effects of aluminium (Al) on Picea abies (L.) Karst. (Norway spruce) trees are well documented in laboratory-scale experiments, but field-based evidence is scarce. This paper presents results on fine root growth and chemistry from a field manipulation experiment in a P. abies stand that was 45 years old when the experiment started in 1996. Different amounts of dissolved aluminium were added as AlCl3 by means of periodic irrigation during the growing season in the period 1997-2002. Potentially toxic concentrations of Al in the soil solution were obtained. Fine roots were studied from direct cores (1996) and sequential root ingrowth cores (1999, 2001, 2002) in the mineral soil (0-40 cm). We tested two hypotheses: (1) elevated concentration of Al in the root zone leads to significant changes in root biomass, partitioning into fine, coarse, living or dead fractions, and distribution with depth; (2) elevated Al concentration leads to a noticeable uptake of Al and reduced uptake of Ca and Mg; this results in Ca and Mg depletion in roots. Hypothesis 1 was only marginally supported, as just a few significant treatment effects on biomass were found. Hypothesis 2 was supported in part; Al addition led to increased root concentrations of Al in 1999 and 2002 and reduced Mg/Al in 1999. Comparison of roots from subsequent root samplings showed a decrease in Al and S over time. The results illustrated that 7 years of elevated Al(tot) concentrations in the soil solution up to 200 microM are not likely to affect root growth. We also discuss possible improvements of the experimental approach.

  13. Analytical approaches to the determination of simple biophenols in forest trees such as Acer (maple), Betula (birch), Coniferus, Eucalyptus, Juniperus (cedar), Picea (spruce) and Quercus (oak).

    Science.gov (United States)

    Bedgood, Danny R; Bishop, Andrea G; Prenzler, Paul D; Robards, Kevin

    2005-06-01

    Analytical methods are reviewed for the determination of simple biophenols in forest trees such as Acer (maple), Betula (birch), Coniferus, Eucalyptus, Juniperus (cedar), Picea (spruce) and Quercus (oak). Data are limited but nevertheless clearly establish the critical importance of sample preparation and pre-treatment in the analysis. For example, drying methods invariably reduce the recovery of biophenols and this is illustrated by data for birch leaves where flavonoid glycosides were determined as 12.3 +/- 0.44 mg g(-1) in fresh leaves but 9.7 +/- 0.35 mg g(-1) in air-dried samples (data expressed as dry weight). Diverse sample handling procedures have been employed for recovery of biophenols. The range of biophenols and diversity of sample types precludes general procedural recommendations. Caution is necessary in selecting appropriate procedures as the high reactivity of these compounds complicates their analysis. Moreover, our experience suggests that their reactivity is very dependent on the matrix. The actual measurement is less contentious and high performance separation methods particularly liquid chromatography dominate analyses whilst coupled techniques involving electrospray ionization are becoming routine particularly for qualitative applications. Quantitative data are still the exception and are summarized for representative species that dominate the forest canopy of various habitats. Reported concentrations for simple phenols range from trace level (<0.1 microg g(-1)) to in excess of 500 microg g(-1) depending on a range of factors. Plant tissue is one of these variables but various biotic and abiotic processes such as stress are also important considerations.

  14. GC/MS Analysis of Oil Extractives from Wood and Bark of Pinus sylvestris, Abies alba, Picea abies, and Larix decidua

    Directory of Open Access Journals (Sweden)

    Mohamed Zidan Mohamed Salem

    2015-09-01

    Full Text Available Wood and bark oil extractives components (OECs of Pinus sylvestris, Abies alba, Picea abies, and Larix decidua grown in the Czech Republic were analyzed using gas chromatography/ mass spectrometry (GC/MS. The analysis showed the presence of monoterpene, sesquiterpene, diterpenoids, and resin acids. The highest percentages of OECs in the wood of P. sylvestris were α-fenchyl alcohol (26.04%, D-fenchyl alcohol (12.39%, and L-borneol (8.81%; the OECs in the bark included α-methyl-γ-butyrolactone (31.88% and isodecyl octyl phthalate (15.85%. The most frequently occurring OEC in A. alba wood were 4-hydroxy-4-methyl-2-pentanone (73.36%, α-cedrol (10.08%, and 2,6-dimethyl-1,3,6-heptatriene (7.35%; the most OECs in the bark were di(2-ethylhexylphthalate (59.83%, methyl cyclopentane (16.63%, and 13-epimanool (6.31%. P. abies wood OECs included 4-hydroxy-4-methyl-2-pentanone (29.42%, α-cedrol (26.98%, ∆3-carene (6.08%, and terpinen-4-ol (5.42%; the most OECs in the bark were di(2-ethylhexylphthalate (30.91%, cyclohexane (12.89%, caryophyllene oxide (8.90%, and α-pinene (4.59%. OECs of L. decidua wood were α-terpineol (26.06%, isoborneol (14.12%, camphene (11.78%, D-fenchyl alcohol (10.39%, and larixol (4.85%; OECs in the bark were larixol (33.29%, phthalic acid mono-2-ethylhexyl ester (16.96%, 13-epimanool (15.40%, and cyclohexane (8.44%.

  15. Variation in light-intercepting area and photosynthetic rate of sun and shade shoots of two Picea species in relation to the angle of incoming light.

    Science.gov (United States)

    Ishii, Hiroaki; Hamada, Yoko; Utsugi, Hajime

    2012-10-01

    We investigated the effects of sun- and shade-shoot architecture on the photosynthetic rates of two Picea species by applying light from various angles in the laboratory. Compared with sun shoots, shade shoots were characterized by lower mass allocation per light-intercepting area, less leaf mass per shoot mass, less mutual shading among leaves and more efficient allocation of chlorophyll to photosynthesis. The shoot silhouette to total leaf area ratio (STAR(ϕ)) decreased with increasing shoot inclination angle (ϕ, the shoot axis angle relative to the projection plane) and was consistently higher for the shade shoots. Morphological and physiological characteristics of the shade shoots resulted in maximum rates of net photosynthesis at ϕ = 0° (P(max,0)) similar to that of the sun shoots when expressed on a leaf mass, total leaf area and chlorophyll basis. When the angle of incoming light was varied, P(max,ϕ) per total leaf area (P(max,ϕ )/A(T)) of the shade shoots increased linearly with increasing STAR(ϕ), while P(max,ϕ) per shoot silhouette area did not change. In contrast, the response of the sun shoots was non-linear, and an optimum angle of incoming light was determined. Our results suggest that shade-shoot morphology is adaptive for utilizing diffuse light incoming from various angles, while sun-shoot morphology is adaptive for avoiding the negative effects of strong direct radiation and for enhancing light diffusion into the canopy. We propose that the angle of incoming light should be taken into account when estimating photosynthetic rates of sun shoots of conifer trees in the field.

  16. BAC and RNA sequencing reveal the brown planthopper resistance gene BPH15 in a recombination cold spot that mediates a unique defense mechanism.

    Science.gov (United States)

    Lv, Wentang; Du, Ba; Shangguan, Xinxin; Zhao, Yan; Pan, Yufang; Zhu, Lili; He, Yuqing; He, Guangcun

    2014-08-11

    Brown planthopper (BPH, Nilaparvata lugens Stål), is the most destructive phloem-feeding insect pest of rice (Oryza sativa). The BPH-resistance gene BPH15 has been proved to be effective in controlling the pest and widely applied in rice breeding programs. Nevertheless, molecular mechanism of the resistance remain unclear. In this study, we narrowed down the position of BPH15 on chromosome 4 and investigated the transcriptome of BPH15 rice after BPH attacked. We analyzed 13,000 BC2F2 plants of cross between susceptible rice TN1 and the recombinant inbred line RI93 that carrying the BPH15 gene from original resistant donor B5. BPH15 was mapped to a 0.0269 cM region on chromosome 4, which is 210-kb in the reference genome of Nipponbare. Sequencing bacterial artificial chromosome (BAC) clones that span the BPH15 region revealed that the physical size of BPH15 region in resistant rice B5 is 580-kb, much bigger than the corresponding region in the reference genome of Nipponbare. There were 87 predicted genes in the BPH15 region in resistant rice. The expression profiles of predicted genes were analyzed. Four jacalin-related lectin proteins genes and one LRR protein gene were found constitutively expressed in resistant parent and considered the candidate genes of BPH15. The transcriptomes of resistant BPH15 introgression line and the susceptible recipient line were analyzed using high-throughput RNA sequencing. In total, 2,914 differentially expressed genes (DEGs) were identified. BPH-responsive transcript profiles were distinct between resistant and susceptible plants and between the early stage (6 h after infestation, HAI) and late stage (48 HAI). The key defense mechanism was related to jasmonate signaling, ethylene signaling, receptor kinase, MAPK cascades, Ca(2+) signaling, PR genes, transcription factors, and protein posttranslational modifications. Our work combined BAC and RNA sequencing to identify candidate genes of BPH15 and revealed the resistance mechanism

  17. Proceso para transferir el conocimiento en la adquisición de software en la Gerencia de Tecnologías de Información del BAC San José

    OpenAIRE

    Castillo Rivera, Roberto

    2016-01-01

    Proyecto final de graduación, Maestría profesional El presente proyecto es el resultado de la aplicación de las mejores prácticas tomando como base la metodología de ITIL en su versión 3 para la liberación final de productos de software, específicamente en cuanto a la transferencia del conocimiento obtenido durante el proceso de adquisición de software aplicativo en la Gerencia de Tecnologías de Información del Banco BAC San José. El Banco BAC San José es una entidad financiera privada,...

  18. The sensitivity of direct identification from positive BacT/ALERT™ (bioMérieux) blood culture bottles by matrix-assisted laser desorption ionization time-of-flight mass spectrometry is low.

    Science.gov (United States)

    Szabados, F; Michels, M; Kaase, M; Gatermann, S

    2011-02-01

    Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been presented as a novel method for the direct identification of bacteria from positive blood culture bottles. The rate of the MALDI TOF MS-based identification in the present study from positive BacT/ALERT (bioMérieux, Marcy l'Etoile, France) blood culture bottles was 30%, which is far below the previously reported sensitivities using the BACTEC (Becton Dickinson, Franklin Lakes, NJ, USA) system. We also found evidence that the Biotyper algorithm did not identify a second pathogen in cases of positive BacT/ALERT blood culture bottles containing two different species. © 2010 The Authors. Journal Compilation © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  19. An evaluation of three processing methods and the effect of reduced culture times for faster direct identification of pathogens from BacT/ALERT blood cultures by MALDI-TOF MS.

    Science.gov (United States)

    Loonen, A J M; Jansz, A R; Stalpers, J; Wolffs, P F G; van den Brule, A J C

    2012-07-01

    Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is a fast and reliable method for the identification of bacteria from agar media. Direct identification from positive blood cultures should decrease the time to obtaining the result. In this study, three different processing methods for the rapid direct identification of bacteria from positive blood culture bottles were compared. In total, 101 positive aerobe BacT/ALERT bottles were included in this study. Aliquots from all bottles were used for three bacterial processing methods, i.e. the commercially available Bruker's MALDI Sepsityper kit, the commercially available Molzym's MolYsis Basic5 kit and a centrifugation/washing method. In addition, the best method was used to evaluate the possibility of MALDI application after a reduced incubation time of 7 h of Staphylococcus aureus- and Escherichia coli-spiked (1,000, 100 and 10 colony-forming units [CFU]) aerobe BacT/ALERT blood cultures. Sixty-six (65%), 51 (50.5%) and 79 (78%) bottles were identified correctly at the species level when the centrifugation/washing method, MolYsis Basic 5 and Sepsityper were used, respectively. Incorrect identification was obtained in 35 (35%), 50 (49.5%) and 22 (22%) bottles, respectively. Gram-positive cocci were correctly identified in 33/52 (64%) of the cases. However, Gram-negative rods showed a correct identification in 45/47 (96%) of all bottles when the Sepsityper kit was used. Seven hours of pre-incubation of S. aureus- and E. coli-spiked aerobe BacT/ALERT blood cultures never resulted in reliable identification with MALDI-TOF MS. Sepsityper is superior for the direct identification of microorganisms from aerobe BacT/ALERT bottles. Gram-negative pathogens show better results compared to Gram-positive bacteria. Reduced incubation followed by MALDI-TOF MS did not result in faster reliable identification.

  20. Comparative analysis of chimeric ZFP-, TALE- and Cas9-piggyBac transposases for integration into a single locus in human cells.

    Science.gov (United States)

    Luo, Wentian; Galvan, Daniel L; Woodard, Lauren E; Dorset, Dan; Levy, Shawn; Wilson, Matthew H

    2017-08-21

    Integrating DNA delivery systems hold promise for many applications including treatment of diseases; however, targeted integration is needed for improved safety. The piggyBac (PB) transposon system is a highly active non-viral gene delivery system capable of integrating defined DNA segments into host chromosomes without requiring homologous recombination. We systematically compared four different engineered zinc finger proteins (ZFP), four transcription activator-like effector proteins (TALE), CRISPR associated protein 9 (SpCas9) and the catalytically inactive dSpCas9 protein fused to the amino-terminus of the transposase enzyme designed to target the hypoxanthine phosphoribosyltransferase (HPRT) gene located on human chromosome X. Chimeric transposases were evaluated for expression, transposition activity, chromatin immunoprecipitation at the target loci, and targeted knockout of the HPRT gene in human cells. One ZFP-PB and one TALE-PB chimera demonstrated notable HPRT gene targeting. In contrast, Cas9/dCas9-PB chimeras did not result in gene targeting. Instead, the HPRT locus appeared to be protected from transposon integration. Supplied separately, PB permitted highly efficient isolation of Cas9-mediated knockout of HPRT, with zero transposon integrations in HPRT by deep sequencing. In summary, these tools may allow isolation of 'targeted-only' cells, be utilized to protect a genomic locus from transposon integration, and enrich for Cas9-mutated cells. Published by Oxford University Press on behalf of Nucleic Acids Research 2017.

  1. Clustered regulatory interspaced short palindromic repeats (CRISPR)-mediated mutagenesis and phenotype rescue by piggyBac transgenesis in a nonmodel Drosophila species.

    Science.gov (United States)

    Tanaka, R; Murakami, H; Ote, M; Yamamoto, D

    2016-08-01

    How behavioural diversity emerged in evolution is an unexplored subject in biology. To tackle this problem, genes and circuits for a behaviour need to be determined in different species for phylogenetic comparisons. The recently developed clustered regulatory interspaced short palindromic repeats/CRISPR associated protein9 (CRISPR/Cas9) system made such a challenge possible by providing the means to induce mutations in a gene of interest in any organism. Aiming at elucidating diversification in genetic and neural networks for courtship behaviour, we attempted to generate a genetic tool kit in Drosophila subobscura, a nonmodel species distantly related to the genetic model Drosophila melanogaster. Here we report the generation of yellow (y) and white mutations with the aid of the CRISPR/Cas9 system, and the rescue of the y mutant phenotype by germline transformation of the newly established y mutant fly line with a y(+) -marked piggyBac vector. This successful mutagenesis and transformation in D. subobscura open up an avenue for comprehensive genetic analyses of higher functions in this and other nonmodel Drosophila species, representing a key step toward systematic comparisons of genes and circuitries underlying behaviour amongst species. © 2016 The Royal Entomological Society.

  2. Detection of an inversion in the Ty-2 region between S. lycopersicum and S. habrochaites by a combination of de novo genome assembly and BAC cloning.

    Science.gov (United States)

    Wolters, Anne-Marie A; Caro, Myluska; Dong, Shufang; Finkers, Richard; Gao, Jianchang; Visser, Richard G F; Wang, Xiaoxuan; Du, Yongchen; Bai, Yuling

    2015-10-01

    A chromosomal inversion associated with the tomato Ty - 2 gene for TYLCV resistance is the cause of severe suppression of recombination in a tomato Ty - 2 introgression line. Among tomato and its wild relatives inversions are often observed, which result in suppression of recombination. Such inversions hamper the transfer of important traits from a related species to the crop by introgression breeding. Suppression of recombination was reported for the TYLCV resistance gene, Ty-2, which has been introgressed in cultivated tomato (Solanum lycopersicum) from the wild relative S. habrochaites accession B6013. Ty-2 was mapped to a 300-kb region on the long arm of chromosome 11. The suppression of recombination in the Ty-2 region could be caused by chromosomal rearrangements in S. habrochaites compared with S. lycopersicum. With the aim of visualizing the genome structure of the Ty-2 region, we compared the draft de novo assembly of S. habrochaites accession LYC4 with the sequence of cultivated tomato ('Heinz'). Furthermore, using populations derived from intraspecific crosses of S. habrochaites accessions, the order of markers in the Ty-2 region was studied. Results showed the presence of an inversion of approximately 200 kb in the Ty-2 region when comparing S. lycopersicum and S. habrochaites. By sequencing a BAC clone from the Ty-2 introgression line, one inversion breakpoint was identified. Finally, the obtained results are discussed with respect to introgression breeding and the importance of a priori de novo sequencing of the species involved.

  3. Direct identification of bacteria from positive BacT/ALERT blood culture bottles using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry.

    Science.gov (United States)

    Mestas, Javier; Felsenstein, Susanna; Bard, Jennifer Dien

    2014-11-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry is a fast and robust method for the identification of bacteria. In this study, we evaluate the performance of a laboratory-developed lysis method (LDT) for the rapid identification of bacteria from positive BacT/ALERT blood culture bottles. Of the 168 positive bottles tested, 159 were monomicrobial, the majority of which were Gram-positive organisms (61.0% versus 39.0%). Using a cut-off score of ≥1.7, 80.4% of the organisms were correctly identified to the species level, and the identification rate of Gram-negative organisms (90.3%) was found to be significantly greater than that of Gram-positive organisms (78.4%). The simplicity and cost-effectiveness of the LDT enable it to be fully integrated into the routine workflow of the clinical microbiology laboratory, allowing for rapid identification of Gram-positive and Gram-negative bacteria within an hour of blood culture positivity. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. High hydrostatic pressure effects on Listeria monocytogenes and L. innocua: Evidence for variability in inactivation behaviour and in resistance to pediocin bacHA-6111-2.

    Science.gov (United States)

    Bruschi, Carolina; Komora, Norton; Castro, Sónia Marília; Saraiva, Jorge; Ferreira, Vânia Borges; Teixeira, Paula

    2017-06-01

    The effect of high hydrostatic pressure (HHP) on the survival of 14 strains of Listeria monocytogenes from food or clinical origins, selected to represent different pheno and genotypes, was evaluated. Stationary phase cells were submitted to 300, 400 and 500 MPa at 10 °C, for 5 min. A high variability in the resistance of L. monocytogenes to pressure was observed, and particularly two strains isolated from food were significantly more baroresistant than the rest. Strains of L. monocytogenes resistant to one or more antibiotics exhibited significantly higher levels of survival after the high pressure treatment at 400 MPa. No correlation was found between strains' origin or thermal tolerance and resistance to HHP. The suitability of two strains of L. innocua as surrogates of L. monocytogenes, was also investigated. These exhibited significantly higher sensitivities to HHP than observed for some L. monocytogenes. The antimicrobial effect of the antilisterial bacteriocin (bacHA-6111-2) increased after L. monocytogenes cells had been exposed to pressure. The data obtained underlines the importance of strain selection for studies aiming to evaluate HHP efficacy to ensure safety of HHP-treated foods. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Genome-wide comparison of paired fresh frozen and formalin-fixed paraffin-embedded gliomas by custom BAC and oligonucleotide array comparative genomic hybridization: facilitating analysis of archival gliomas.

    Science.gov (United States)

    Mohapatra, Gayatry; Engler, David A; Starbuck, Kristen D; Kim, James C; Bernay, Derek C; Scangas, George A; Rousseau, Audrey; Batchelor, Tracy T; Betensky, Rebecca A; Louis, David N

    2011-04-01

    Array comparative genomic hybridization (aCGH) is a powerful tool for detecting DNA copy number alterations (CNA). Because diffuse malignant gliomas are often sampled by small biopsies, formalin-fixed paraffin-embedded (FFPE) blocks are often the only tissue available for genetic analysis; FFPE tissues are also needed to study the intratumoral heterogeneity that characterizes these neoplasms. In this paper, we present a combination of evaluations and technical advances that provide strong support for the ready use of oligonucleotide aCGH on FFPE diffuse gliomas. We first compared aCGH using bacterial artificial chromosome (BAC) arrays in 45 paired frozen and FFPE gliomas, and demonstrate a high concordance rate between FFPE and frozen DNA in an individual clone-level analysis of sensitivity and specificity, assuring that under certain array conditions, frozen and FFPE DNA can perform nearly identically. However, because oligonucleotide arrays offer advantages to BAC arrays in genomic coverage and practical availability, we next developed a method of labeling DNA from FFPE tissue that allows efficient hybridization to oligonucleotide arrays. To demonstrate utility in FFPE tissues, we applied this approach to biphasic anaplastic oligoastrocytomas and demonstrate CNA differences between DNA obtained from the two components. Therefore, BAC and oligonucleotide aCGH can be sensitive and specific tools for detecting CNAs in FFPE DNA, and novel labeling techniques enable the routine use of oligonucleotide arrays for FFPE DNA. In combination, these advances should facilitate genome-wide analysis of rare, small and/or histologically heterogeneous gliomas from FFPE tissues.

  6. Comparative physical mapping of rice BAC clones linked to resistance genes Glh,Bph-3 and xa-5 in Oryza sativa L.and O.granulata Nees et Am.ex Watt.

    Institute of Scientific and Technical Information of China (English)

    XIONG Zhiyong; TAN Guangxuan; YOU Aiqing; HE Guangyuan; SHE Chaowen; LI Lijia; SONG Yunchun

    2004-01-01

    Oryza granulata Nees et Arn. ex Watt. is one of the three wild relatives of rice, which are the most valuable for study and utilization in China. In this study, the homology and physical locations of three rice resistance genes, Glh,Bph-3 and xa-5 are comparatively analyzed between O. sativa and O. granulata by Southern blotting and fluorescence in situ hybridization (FISH). The results of Southern blotting indicate that there exist homologous sequences of the tested RFLP markers in O. granulata. By using three bacterial artificial chromosome (BAC) clones scanned by the tested RFLP as probes, FISH signals are detected on both mitotic and pachytene chromosomes in O. sativa and O. granulata.Dual-color FISH demonstrates that two of the three BAC clones (14E16 and 38J9) are located on the short arm of the same chromosome pair in O. granulata. Additionally, colinearity is shown for the two clones between O. sativa and O.granulata. Another BAC clone 44B4 is located on the end of the short arm of other chromosome pair in these two species.Although the phylogenetic relationship between O. sativa and O. granulata is the most distinct in Oryza and these two species have evidently different biological features and ecological habits, the relative lengths and arm ratios of the detected chromosomes and the relative positions of the tested clone signals on chromosomes in O. granulata are quite similar to those in O. sativa.

  7. Factors affecting the accuracy of genomic selection for growth and wood quality traits in an advanced-breeding population of black spruce (Picea mariana).

    Science.gov (United States)

    Lenz, Patrick R N; Beaulieu, Jean; Mansfield, Shawn D; Clément, Sébastien; Desponts, Mireille; Bousquet, Jean

    2017-04-28

    Genomic selection (GS) uses information from genomic signatures consisting of thousands of genetic markers to predict complex traits. As such, GS represents a promising approach to accelerate tree breeding, which is especially relevant for the genetic improvement of boreal conifers characterized by long breeding cycles. In the present study, we tested GS in an advanced-breeding population of the boreal black spruce (Picea mariana [Mill.] BSP) for growth and wood quality traits, and concurrently examined factors affecting GS model accuracy. The study relied on 734 25-year-old trees belonging to 34 full-sib families derived from 27 parents and that were established on two contrasting sites. Genomic profiles were obtained from 4993 Single Nucleotide Polymorphisms (SNPs) representative of as many gene loci distributed among the 12 linkage groups common to spruce. GS models were obtained for four growth and wood traits. Validation using independent sets of trees showed that GS model accuracy was high, related to trait heritability and equivalent to that of conventional pedigree-based models. In forward selection, gains per unit of time were three times higher with the GS approach than with conventional selection. In addition, models were also accurate across sites, indicating little genotype-by-environment interaction in the area investigated. Using information from half-sibs instead of full-sibs led to a significant reduction in model accuracy, indicating that the inclusion of relatedness in the model contributed to its higher accuracies. About 500 to 1000 markers were sufficient to obtain GS model accuracy almost equivalent to that obtained with all markers, whether they were well spread across the genome or from a single linkage group, further confirming the implication of relatedness and potential long-range linkage disequilibrium (LD) in the high accuracy estimates obtained. Only slightly higher model accuracy was obtained when using marker subsets that were

  8. Applicability of non-destructive substitutes for leaf area in different stands of Norway spruce (Picea abies L. Karst.) focusing on traditional forest crown measures

    Science.gov (United States)

    Laubhann, Daniel; Eckmüllner, Otto; Sterba, Hubert

    2010-01-01

    Since individual tree leaf area is an important measure for productivity as well as for site occupancy, it is of high interest in many studies about forest growth. The exact determination of leaf area is nearly impossible. Thus, a common way to get information about leaf area is to use substitutes. These substitutes are often variables which are collected in a destructive way which is not feasible for long term studies. Therefore, this study aimed at testing the applicability of using substitutes for leaf area which could be collected in a non-destructive way, namely crown surface area and crown projection area. In 8 stands of Norway spruce (Picea abies L. Karst.), divided into three age classes and two thinning treatments, a total of 156 trees were felled in order to test the relationship between leaf area and crown surface area and crown projection area, respectively. Individual tree leaf area of the felled sample trees was estimated by 3P-branch sampling with an accuracy of ±10%. Crown projection area and crown surface area were compared with other, more commonly used, but destructive predictors of leaf area, namely sapwood area at different heights on the bole. Our investigations confirmed findings of several studies that sapwood area is the most precise measure for leaf area because of the high correlation between sapwood area and the leaf area. But behind sapwood area at crown base and sapwood area at three tenth of the tree height the predictive ability of crown surface area was ranked third and even better than that of sapwood area at breast height (R2 = 0.656 compared with 0.600). Within the stands leaf area is proportional to crown surface area. Using the pooled data of all stands a mixed model approach showed that additionally to crown surface area dominant height and diameter at breast height (dbh) improved the leaf area estimates. Thus, taking dominant height and dbh into account, crown surface area can be recommended for estimating the leaf area of

  9. Age-related effects on leaf area/sapwood area relationships, canopy transpiration and carbon gain of Norway spruce stands (Picea abies) in the Fichtelgebirge, Germany.

    Science.gov (United States)

    Köstner, B; Falge, E; Tenhunen, J D

    2002-06-01

    Stand age is an important structural determinant of canopy transpiration (E(c)) and carbon gain. Another more functional parameter of forest structure is the leaf area/sapwood area relationship, A(L)/A(S), which changes with site conditions and has been used to estimate leaf area index of forest canopies. The interpretation of age-related changes in A(L)/A(S) and the question of how A(L)/A(S) is related to forest functions are of current interest because they may help to explain forest canopy fluxes and growth. We conducted studies in mature stands of Picea abies (L.) Karst. varying in age from 40 to 140 years, in tree density from 1680 to 320 trees ha(-1), and in tree height from 15 to 30 m. Structural parameters were measured by biomass harvests of individual trees and stand biometry. We estimated E(c) from scaled-up xylem sap flux of trees, and canopy-level fluxes were predicted by a three-dimensional microclimate and gas exchange model (STANDFLUX). In contrast to pine species, A(L)/A(S) of P. abies increased with stand age from 0.26 to 0.48 m(2) cm(-2). Agreement between E(c) derived from scaled-up sap flux and modeled canopy transpiration was obtained with the same parameterization of needle physiology independent of stand age. Reduced light interception per leaf area and, as a consequence, reductions in net canopy photosynthesis (A(c)), canopy conductance (g(c)) and E(c) were predicted by the model in the older stands. Seasonal water-use efficiency (WUE = A(c)/E(c)), derived from scaled-up sap flux and stem growth as well as from model simulation, declined with increasing A(L)/A(S) and stand age. Based on the different behavior of age-related A(L)/A(S) in Norway spruce stands compared with other tree species, we conclude that WUE rather than A(L)/A(S) could represent a common age-related property of all species. We also conclude that, in addition to hydraulic limitations reducing carbon gain in old stands, a functional change in A(L)/A(S) that is related to

  10. Applicability of non-destructive substitutes for leaf area in different stands of Norway spruce (Picea abies L. Karst.) focusing on traditional forest crown measures.

    Science.gov (United States)

    Laubhann, Daniel; Eckmüllner, Otto; Sterba, Hubert

    2010-09-30

    Since individual tree leaf area is an important measure for productivity as well as for site occupancy, it is of high interest in many studies about forest growth. The exact determination of leaf area is nearly impossible. Thus, a common way to get information about leaf area is to use substitutes. These substitutes are often variables which are collected in a destructive way which is not feasible for long term studies. Therefore, this study aimed at testing the applicability of using substitutes for leaf area which could be collected in a non-destructive way, namely crown surface area and crown projection area. In 8 stands of Norway spruce (Picea abies L. Karst.), divided into three age classes and two thinning treatments, a total of 156 trees were felled in order to test the relationship between leaf area and crown surface area and crown projection area, respectively. Individual tree leaf area of the felled sample trees was estimated by 3P-branch sampling with an accuracy of ±10%. Crown projection area and crown surface area were compared with other, more commonly used, but destructive predictors of leaf area, namely sapwood area at different heights on the bole. Our investigations confirmed findings of several studies that sapwood area is the most precise measure for leaf area because of the high correlation between sapwood area and the leaf area. But behind sapwood area at crown base and sapwood area at three tenth of the tree height the predictive ability of crown surface area was ranked third and even better than that of sapwood area at breast height (R(2) = 0.656 compared with 0.600). Within the stands leaf area is proportional to crown surface area. Using the pooled data of all stands a mixed model approach showed that additionally to crown surface area dominant height and diameter at breast height (dbh) improved the leaf area estimates. Thus, taking dominant height and dbh into account, crown surface area can be recommended for estimating the leaf area

  11. Mechanical Properties of Picea sitchensis

    DEFF Research Database (Denmark)

    Bräuner, Lise; Hoffmeyer, Preben; Poulsson, Lise

    2000-01-01

    the requirements at the same level as Danish grown Norway spruce. The study shows that Sitka spruce and Norway spruce of the same origin exhibit highly comparable mechanical properties. Key words: annual ring width, bending strength, characteristic strength, dry density, EN 338, INSTA 142, modulus of elasticity...

  12. Tumor-directed gene therapy in mice using a composite nonviral gene delivery system consisting of the piggyBac transposon and polyethylenimine

    International Nuclear Information System (INIS)

    Kang, Yu; Zhang, Xiaoyan; Jiang, Wei; Wu, Chaoqun; Chen, Chunmei; Zheng, Yufang; Gu, Jianren; Xu, Congjian

    2009-01-01

    Compared with viral vectors, nonviral vectors are less immunogenic, more stable, safer and easier to replication for application in cancer gene therapy. However, nonviral gene delivery system has not been extensively used because of the low transfection efficiency and the short transgene expression, especially in vivo. It is desirable to develop a nonviral gene delivery system that can support stable genomic integration and persistent gene expression in vivo. Here, we used a composite nonviral gene delivery system consisting of the piggyBac (PB) transposon and polyethylenimine (PEI) for long-term transgene expression in mouse ovarian tumors. A recombinant plasmid PB [Act-RFP, HSV-tk] encoding both the herpes simplex thymidine kinase (HSV-tk) and the monomeric red fluorescent protein (mRFP1) under PB transposon elements was constructed. This plasmid and the PBase plasmid were injected into ovarian cancer tumor xenografts in mice by in vivo PEI system. The antitumor effects of HSV-tk/ganciclovir (GCV) system were observed after intraperitoneal injection of GCV. Histological analysis and TUNEL assay were performed on the cryostat sections of the tumor tissue. Plasmid construction was confirmed by PCR analysis combined with restrictive enzyme digestion. mRFP1 expression could be visualized three weeks after the last transfection of pPB/TK under fluorescence microscopy. After GCV admission, the tumor volume of PB/TK group was significantly reduced and the tumor inhibitory rate was 81.96% contrasted against the 43.07% in the TK group. Histological analysis showed that there were extensive necrosis and lymphocytes infiltration in the tumor tissue of the PB/TK group but limited in the tissue of control group. TUNEL assays suggested that the transfected cells were undergoing apoptosis after GCV admission in vivo. Our results show that the nonviral gene delivery system coupling PB transposon with PEI can be used as an efficient tool for gene therapy in ovarian cancer

  13. Superior sensitivity and decreased time to detection with the Bactec Peds Plus/F system compared to the BacT/Alert Pediatric FAN blood culture system.

    Science.gov (United States)

    Sullivan, K V; Turner, N N; Lancaster, D P; Shah, A R; Chandler, L J; Friedman, D F; Blecker-Shelly, D L

    2013-12-01

    Here, we compare the sensitivities and times to detection (TTD) of BacT/Alert Pediatric FAN (PF) and Bactec Peds Plus blood culture bottles. Test bottles were inoculated with 2 ml of banked whole blood, 1-ml aliquots of antibiotic suspension, and organisms diluted to simulate a bacteremia level of 10 to 100 CFU/ml. The control bottles were inoculated with 3 ml of banked blood and organism suspensions only. The organism-drug combinations were Staphylococcus epidermidis and vancomycin, methicillin-resistant Staphylococcus aureus and vancomycin, Streptococcus pneumoniae, vancomycin, and ceftriaxone, Streptococcus agalactiae, ampicillin, and cefotaxime, Escherichia coli, cefotaxime, and cefepime, Pseudomonas aeruginosa, piperacillin-tazobactam, cefepime, and gentamicin, Neisseria meningitidis and ceftriaxone, and Haemophilus influenzae and ceftriaxone. The control and test bottle combinations were tested in duplicate. The bottles were incubated for 5 days; 32 control and 104 test bottles were incubated. Overall, the bacterial recovery rates for the PF and Peds Plus bottles were 37% and 62%, 94% and 100% in the controls, 19% and 50% in the test bottles, and 33% and 92% in the bottles with vancomycin, respectively. No bacteria were recovered from the bottles with S. pneumoniae, S. agalactiae, E. coli, N. meningitidis, or H. influenzae in combination with cefotaxime or ceftriaxone. The Peds Plus system detected P. aeruginosa in bottles with cefepime and piperacillin-tazobactam, but the PF system recovered bacteria only in bottles with trough levels of piperacillin-tazobactam. The mean TTD were shorter in the Peds Plus system controls (14.2 versus 18.0 h; P = 0.001) and the test bottles (14.3 versus 17.8 h; P = 0.008) than in the PF bottles. Overall, we demonstrated superior sensitivity, TTD, and antibiotic neutralization in the Bactec Peds Plus system compared to those in the Pediatric FAN system.

  14. Spatial distribution of lead and lead isotopes in soil B-horizon, forest-floor humus, grass (Avenella flexuosa) and spruce (Picea abies) needles across the Czech Republic

    Energy Technology Data Exchange (ETDEWEB)

    Sucharova, Julie; Suchara, Ivan [Silva Tarouca Research Institute for Landscape and Ornamental Gardening, Kvetnove namesti 391, 252 43 Pruhonice (Czech Republic); Reimann, Clemens, E-mail: Clemens.Reimann@ngu.no [Geological Survey of Norway, P.O. Box 6315 Sluppen, 7491 Trondheim (Norway); Boyd, Rognvald [Geological Survey of Norway, P.O. Box 6315 Sluppen, 7491 Trondheim (Norway); Filzmoser, Peter [Institute for Statistics and Probability Theory, Vienna University of Technology, Wiedner Hauptstrasse 8-10, 1040 Wien (Austria); Englmaier, Peter [Faculty of Life Science, University of Vienna, Althanstr. 14, A-1090 Vienna (Austria)

    2011-07-15

    Highlights: > Pb-concentrations and {sup 206}Pb/{sup 207}Pb isotope ratios are provided for four different sample materials for the Czech Republic. > The paper demonstrates the local impact of a number of different contamination sources. > The data provide clear evidence that traffic emissions are no major source of Pb to the Czech environment. > The data demonstrate that the B-horizon provides no valid 'background' for Pb-concentration or the {sup 206}Pb/{sup 207}Pb isotope ratio. > Pb isotope ratios change during soil weathering and at the interface biosphere/pedosphere. - Abstract: Lead concentrations were determined in samples of soil B-horizon (N = 258), forest-floor humus (O-horizon, N = 259), grass (Avenella flexuosa, N = 251) and spruce (Picea abies, N = 253) needles (2nd year) collected at the same locations evenly spread over the territory of the Czech Republic at an average density of 1 site/300 km{sup 2}. Median Pb concentrations differ widely in the four materials: soil B-horizon: 27 mg/kg (3.3-220 mg/kg), humus: 78 mg/kg (19-1863 mg/kg), grass: 0.37 mg/kg (0.08-8 mg/kg) and spruce needles: 0.23 mg/kg (0.07-3 mg/kg). In the Pb distribution maps for humus, grass and spruce a number of well-known Pb-contamination sources are indicated by unusually high concentrations (e.g., the Pb smelter at Pribram, the metallurgical industry in the NE of the Czech Republic and along the Polish border, as well as the metallurgical industry in Upper Silesia and Europe's largest coal-fired power plant at Bogatynia, Poland). The ratio {sup 206}Pb/{sup 207}Pb was determined in all four materials. The median value of the {sup 206}Pb/{sup 207}Pb isotope ratio in the soil B-horizon is 1.184 (variation: 1.145-1.337). In both humus and grass the median value for the {sup 206}Pb/{sup 207}Pb isotope ratio is 1.162 (variation: 1.130-1.182), in spruce needles the median ratio is 1.159 (variation: 1.116-1.186). In humus, grass and spruce needles the known contamination

  15. Allometry of Sapwood Depth in Five Boreal Trees

    OpenAIRE

    M. Rebeca Quiñonez-Piñón; Caterina Valeo

    2017-01-01

    This paper analyzes sapwood variability and allometry within individuals of Populus tremuloides, Pinus contorta, Pinus banksiana, Picea mariana, and Picea glauca. Outside bark diameter at breast height (DBH) and sapwood depth (sd) in four cardinal directions were measured in individuals in stands in Alberta and Saskatchewan, Canada. The microscopical analysis of wood anatomy was used to measure sd, and the error associated with the measures was observed. Sapwood allometry analyses examined th...

  16. Performance of two resin-containing blood culture media in detection of bloodstream infections and in direct matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) broth assays for isolate identification: clinical comparison of the BacT/Alert Plus and Bactec Plus systems.

    Science.gov (United States)

    Fiori, Barbara; D'Inzeo, Tiziana; Di Florio, Viviana; De Maio, Flavio; De Angelis, Giulia; Giaquinto, Alessia; Campana, Lara; Tanzarella, Eloisa; Tumbarello, Mario; Antonelli, Massimo; Sanguinetti, Maurizio; Spanu, Teresa

    2014-10-01

    We compared the clinical performances of the BacT/Alert Plus (bioMérieux) and Bactec Plus (Becton Dickinson) aerobic and anaerobic blood culture (BC) media with adsorbent polymeric beads. Patients ≥ 16 years old with suspected bloodstream infections (BSIs) were enrolled in intensive care units and infectious disease wards. A single 40-ml blood sample was collected from each and used to inoculate (10 ml/bottle) one set of BacT/Alert Plus cultures and one set of Bactec Plus cultures, each set consisting of one aerobic and one anaerobic bottle. Cultures were incubated ≤ 5 days in the BacT/Alert 3D and Bactec FX instruments, respectively. A total of 128 unique BSI episodes were identified based on the recovery of clinically significant growth in 212 aerobic cultures (106 BacT/Alert and 106 Bactec) and 151 anaerobic cultures (82 BacT/Alert and 69 Bactec). The BacT/Alert aerobic medium had higher recovery rates for Gram-positive cocci (P = 0.024), whereas the Bactec aerobic medium was superior for recovery of Gram-negative bacilli (P = 0.006). BacT/Alert anaerobic medium recovery rates exceeded those of the Bactec anaerobic medium for total organisms (P = 0.003), Gram-positive cocci (P = 0.013), and Escherichia coli (P = 0.030). In terms of capacity for diagnosing the 128 septic episodes, the BacT/Alert and Bactec sets were comparable, although the former sets diagnosed more BSIs caused by Gram-positive cocci (P = 0.008). They also allowed earlier identification of coagulase-negative staphylococcal growth (mean, 2.8 h; P = 0.003) and growth in samples from patients not on antimicrobial therapy that yielded positive results (mean, 1.3 h; P direct matrix-assisted laser desorption ionization-time of flight mass spectrometry assay of BC broths. The BacT/Alert Plus media line appears to be a reliable, timesaving tool for routine detection of BSIs in the population we studied, although further studies are needed to evaluate their performance in other settings. Copyright

  17. Amélioration de la politique de sécurité des systèmes DRM introduction des notions du modèle Or-BAC dans le langage MPEG-21 REL

    Directory of Open Access Journals (Sweden)

    Morad Rafi

    2013-12-01

    Full Text Available Digital Rights Management (DRM allows controlling the access, the use and the diffusion of digital contents produced by the rights holders. With this intention, a right expression language is necessary. The MPEG-21 REL is a standardized language used to express the licenses. In this paper, we propose an improvement of the security policy and expressivity of MPEG-21 REL by introducing concepts based on the Or-BAC (Organization Based Access Control data model. This improvement will be illustrated by some use cases

  18. Rapid identification of bacteria from bioMérieux BacT/ALERT blood culture bottles by MALDI-TOF MS.

    Science.gov (United States)

    Haigh, J D; Green, I M; Ball, D; Eydmann, M; Millar, M; Wilks, M

    2013-01-01

    Several studies have reported poor results when trying to identify microorganisms directly from the bioMérieux BacT/ALERT blood culture system using matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry. The aim of this study is to evaluate two new methods, Sepsityper and an enrichment method for direct identification of microorganisms from this system. For both methods the samples were processed using the Bruker Microflex LT mass spectrometer (Biotyper) using the Microflex Control software to obtain spectra. The results from direct analysis were compared with those obtained by subculture and subsequent identification. A total of 350 positive blood cultures were processed simultaneously by the two methods. Fifty-three cultures were polymocrobial or failed to grow any organism on subculture, and these results were not included as there was either no subculture result, or for polymicrobial cultures it was known that the Biotyper would not be able to distinguish the constituent organisms correctly. Overall, the results showed that, contrary to previous reports, it is possible to identify bacteria directly from bioMérieux blood culture bottles, as 219/297 (74%) correct identifications were obtained using the Bruker Sepsityper method and 228/297 (77%) were obtained for the enrichment method when there is only one organism was present. Although the enrichment method was simpler, the reagent costs for the Sepsityper method were approximately pound 4.00 per sample compared to pound 0.50. An even simpler and cheaper method, which was less labour-intensive and did not require further reagents, was investigated. Seventy-seven specimens from positive signalled blood cultures were analysed by inoculating prewarmed blood agar plates and analysing any growth after 1-, 2- and 4-h periods of incubation at 37 degrees C, by either direct transfer or alcohol extraction. This method gave the highest number of correct identifications, 66/77 (86

  19. The Performance of the Four Anaerobic Blood Culture Bottles BacT/ALERT-FN, -FN Plus, BACTEC-Plus and -Lytic in Detection of Anaerobic Bacteria and Identification by Direct MALDI-TOF MS.

    Science.gov (United States)

    Almuhayawi, Mohammed; Altun, Osman; Abdulmajeed, Adam Dilshad; Ullberg, Måns; Özenci, Volkan

    2015-01-01

    Detection and identification of anaerobic bacteria in blood cultures (BC) is a well-recognized challenge in clinical microbiology. We studied 100 clinical anaerobic BC isolates to evaluate the performance of BacT/ALERT-FN, -FN Plus (BioMérieux), BACTEC-Plus and -Lytic (Becton Dickinson BioSciences) BC bottles in detection and time to detection (TTD) of anaerobic bacteria. BACTEC Lytic had higher detection rate (94/100, 94%) than BacT/ALERT FN Plus (80/100, 80%) (panaerobic bacteria among the remaining bottle types. The 67 anaerobic bacteria that signalled positive in all four bottle types were analyzed to compare the time to detection (TTD) and isolates were directly identified by MALDI-TOF MS. There was a significant difference in TTD among the four bottle types (panaerobic BC bottles are equally suitable for direct MALDI-TOF MS for rapid and reliable identification of common anaerobic bacteria. Further clinical studies are warranted to investigate the performance of anaerobic BC bottles in detection of anaerobic bacteria and identification by direct MALDI-TOF MS.

  20. Revisión histórica de los estudios sobre la biología, ecología y toxicología del tiburón azul Prionace glauca en aguas del Pacífico ecuatoriano

    Directory of Open Access Journals (Sweden)

    Calle-Morán, Marcos

    2016-08-01

    Full Text Available Blue shark Prionace glauca is a species that represents a great economic and ecological importance. However, the information related to its biology and others aspects of its population is limited. Nevertheless these kinds of studies are very important for a suitable management of fishery activities. The aim of this research was to develop a historical review for all biological, ecological and toxicological studies made in the coastal and marine ecosystems of the Ecuadorian Pacific. A review of the specializedliteraturewas carried out for many documents such as thesis, technical reports, scientific papers,and others. According to results obtained, in Ecuador there have been four scientific research studies from 2012: These were two of Biology (one of reproductive aspects in Santa Rosa de Salinas and another about age and growth in Manta and the others two of Ecology and Toxicology (one on mercury bioaccumulation and bio-magnification of mercury, and another on the concentration of mercury and cadmium; both conducted in Santa Rosa de Salinas.The works were developed by state and private universities through thesis, while a draft was in preparation for scientific publication as part of a research project in a private institution of higher education

  1. Response of Lutz, Sitka, and white spruce to attack by Dendroctonus rufipennis (Coleoptera: Scolytidae) and blue stain fungi

    Science.gov (United States)

    Richard A. Werner; Barbara L. Illman

    1994-01-01

    Mechanical wounding and wounding plus inoculation with a blue-stain fungus, Leptographium abietinum (Peck), associated with the spruce beetle, Dendroctonus rufipennis (Kirby), caused an induced reaction zone or lesion around the wound sites in Lutz spruce, Picea lutzii Little, Sitka spruce, P. sitchensis (Bong.) Carr., and white spruce, P. glauca (Moench) Voss, in...

  2. Phenology of Lymantria monacha (Lepidoptera:Lymantriidae) laboratory reared on spruce foliage or a newly developed artificial diet

    Science.gov (United States)

    Melody A. Keena; Alice Vandel; Oldrich. Pultar

    2010-01-01

    Lymantria monacha (L.) (Lepidoptera: Lymantriidae) is a Eurasian pest of conifers that has potential for accidental introduction into North America. The phenology over the entire life cycle for L. monacha individuals from the Czech Republic was compared on Picea glauca (Moench) Voss (white spruce) and a newly...

  3. Vulnerability of white spruce tree growth in interior Alaska in response to climate variability: dendrochronological, demographic, and experimental perspectives

    Science.gov (United States)

    A.D. McGuire; R.W. Ruess; A. Lloyd; J. Yarie; J.S. Clein; G.P. Juday

    2010-01-01

    This paper integrates dendrochronological, demographic, and experimental perspectives to improve understanding of the response of white spruce (Picea glauca (Moench) Voss) tree growth to climatic variability in interior Alaska. The dendrochronological analyses indicate that climate warming has led to widespread declines in white spruce growth...

  4. Performance of the Forest Vegetation Simulator in managed white spruce plantations influenced by eastern spruce budworm in northern Minnesota

    Science.gov (United States)

    Matthew B. Russell; Anthony W. D' Amato; Michael A. Albers; Christopher W. Woodall; Klaus J. Puettmann; Michael R. Saunders; Curtis L. VanderSchaaf

    2015-01-01

    Silvicultural strategies such as thinning may minimize productivity losses from a variety of forest disturbances, including forest insects. This study analyzed the 10-year postthinning response of stands and individual trees in thinned white spruce (Picea glauca [Moench] Voss) plantations in northern Minnesota, USA, with light to moderate defoliation...

  5. Compilation of 1992 Annual Reports of the Navy ELF Communications System Ecological Monitoring Program. Volume 2. Tabs C - F

    Science.gov (United States)

    1993-08-01

    Picea glauca, and Pinus resinosus. Centaurea maculosa has increased since 1983 until it is now the most I abundant flowering plant on the hill. Also...population of Centaurea maculosa that bloomed in late July, in spite of the drought and hot temperatures. ThisI plant was not as abundant at the F2

  6. Establishment and growth of white spruce on a boreal forest floodplain: interactions between microclimate and mammalian herbivory

    Science.gov (United States)

    Amy C. Angell; Knut. Kielland

    2009-01-01

    White spruce (Picea glauca (Moench) Voss) is a dominant species in late-successional ecosystems along the Tanana River, interior Alaska, and the most important commercial timber species in these boreal floodplain forests. Whereas white spruce commonly seed in on young terraces in early primary succession, the species does not become a conspicuous...

  7. Dicty_cDB: SHH135 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available and phloem from mature trees Picea glauca cDNA clone GenomeQuebec_Id:GQ00610_L03 5', mRNA sequence. 44 1e-0...RNA, partial cds. 1265 0.0 1 DV977319 |DV977319.1 GQ00610.B3.1_L03 GQ006: Cambium

  8. Tree seed handling, processing, testing, and storage at Hayward State Nursery, Wisconsin Department of Natural Resources

    Science.gov (United States)

    Gordon Christians

    2008-01-01

    The Hayward State Nursery, Wisconsin grows more than 40 species from seeds. Up to 6000 bushels of raw unprocessed tree and shrub seeds are collected each year, and all seeds are collected in Wisconsin or adjacent states. All white spruce (Picea glauca) and some white pine seeds (Pinus strobus) are collected from orchards containing...

  9. Non-functional plastid ndh gene fragments are present in the nuclear genome of Norway spruce (Picea abies L. Karsch): insights from in silico analysis of nuclear and organellar genomes.

    Science.gov (United States)

    Ranade, Sonali Sachin; García-Gil, María Rosario; Rosselló, Josep A

    2016-04-01

    Many genes have been lost from the prokaryote plastidial genome during the early events of endosymbiosis in eukaryotes. Some of them were definitively lost, but others were relocated and functionally integrated to the host nuclear genomes through serial events of gene transfer during plant evolution. In gymnosperms, plastid genome sequencing has revealed the loss of ndh genes from several species of Gnetales and Pinaceae, including Norway spruce (Picea abies). This study aims to trace the ndh genes in the nuclear and organellar Norway spruce genomes. The plastid genomes of higher plants contain 11 ndh genes which are homologues of mitochondrial genes encoding subunits of the proton-pumping NADH-dehydrogenase (nicotinamide adenine dinucleotide dehydrogenase) or complex I (electron transport chain). Ndh genes encode 11 NDH polypeptides forming the Ndh complex (analogous to complex I) which seems to be primarily involved in chloro-respiration processes. We considered ndh genes from the plastidial genome of four gymnosperms (Cryptomeria japonica, Cycas revoluta, Ginkgo biloba, Podocarpus totara) and a single angiosperm species (Arabidopsis thaliana) to trace putative homologs in the nuclear and organellar Norway spruce genomes using tBLASTn to assess the evolutionary fate of ndh genes in Norway spruce and to address their genomic location(s), structure, integrity and functionality. The results obtained from tBLASTn were subsequently analyzed by performing homology search for finding ndh specific conserved domains using conserved domain search. We report the presence of non-functional plastid ndh gene fragments, excepting ndhE and ndhG genes, in the nuclear genome of Norway spruce. Regulatory transcriptional elements like promoters, TATA boxes and enhancers were detected in the upstream regions of some ndh fragments. We also found transposable elements in the flanking regions of few ndh fragments suggesting nuclear rearrangements in those regions. These evidences

  10. Comparative genomic mapping of the bovine Fragile Histidine Triad (FHIT tumour suppressor gene: characterization of a 2 Mb BAC contig covering the locus, complete annotation of the gene, analysis of cDNA and of physiological expression profiles

    Directory of Open Access Journals (Sweden)

    Boussaha Mekki

    2006-05-01

    Full Text Available Abstract Background The Fragile Histidine Triad gene (FHIT is an oncosuppressor implicated in many human cancers, including vesical tumors. FHIT is frequently hit by deletions caused by fragility at FRA3B, the most active of human common fragile sites, where FHIT lays. Vesical tumors affect also cattle, including animals grazing in the wild on bracken fern; compounds released by the fern are known to induce chromosome fragility and may trigger cancer with the interplay of latent Papilloma virus. Results The bovine FHIT was characterized by assembling a contig of 78 BACs. Sequence tags were designed on human exons and introns and used directly to select bovine BACs, or compared with sequence data in the bovine genome database or in the trace archive of the bovine genome sequencing project, and adapted before use. FHIT is split in ten exons like in man, with exons 5 to 9 coding for a 149 amino acids protein. VISTA global alignments between bovine genomic contigs retrieved from the bovine genome database and the human FHIT region were performed. Conservation was extremely high over a 2 Mb region spanning the whole FHIT locus, including the size of introns. Thus, the bovine FHIT covers about 1.6 Mb compared to 1.5 Mb in man. Expression was analyzed by RT-PCR and Northern blot, and was found to be ubiquitous. Four cDNA isoforms were isolated and sequenced, that originate from an alternative usage of three variants of exon 4, revealing a size very close to the major human FHIT cDNAs. Conclusion A comparative genomic approach allowed to assemble a contig of 78 BACs and to completely annotate a 1.6 Mb region spanning the bovine FHIT gene. The findings confirmed the very high level of conservation between human and bovine genomes and the importance of comparative mapping to speed the annotation process of the recently sequenced bovine genome. The detailed knowledge of the genomic FHIT region will allow to study the role of FHIT in bovine cancerogenesis

  11. Comparative genomic mapping of the bovine Fragile Histidine Triad (FHIT) tumour suppressor gene: characterization of a 2 Mb BAC contig covering the locus, complete annotation of the gene, analysis of cDNA and of physiological expression profiles.

    Science.gov (United States)

    Uboldi, Cristina; Guidi, Elena; Roperto, Sante; Russo, Valeria; Roperto, Franco; Di Meo, Giulia Pia; Iannuzzi, Leopoldo; Floriot, Sandrine; Boussaha, Mekki; Eggen, André; Ferretti, Luca

    2006-05-23

    The Fragile Histidine Triad gene (FHIT) is an oncosuppressor implicated in many human cancers, including vesical tumors. FHIT is frequently hit by deletions caused by fragility at FRA3B, the most active of human common fragile sites, where FHIT lays. Vesical tumors affect also cattle, including animals grazing in the wild on bracken fern; compounds released by the fern are known to induce chromosome fragility and may trigger cancer with the interplay of latent Papilloma virus. The bovine FHIT was characterized by assembling a contig of 78 BACs. Sequence tags were designed on human exons and introns and used directly to select bovine BACs, or compared with sequence data in the bovine genome database or in the trace archive of the bovine genome sequencing project, and adapted before use. FHIT is split in ten exons like in man, with exons 5 to 9 coding for a 149 amino acids protein. VISTA global alignments between bovine genomic contigs retrieved from the bovine genome database and the human FHIT region were performed. Conservation was extremely high over a 2 Mb region spanning the whole FHIT locus, including the size of introns. Thus, the bovine FHIT covers about 1.6 Mb compared to 1.5 Mb in man. Expression was analyzed by RT-PCR and Northern blot, and was found to be ubiquitous. Four cDNA isoforms were isolated and sequenced, that originate from an alternative usage of three variants of exon 4, revealing a size very close to the major human FHIT cDNAs. A comparative genomic approach allowed to assemble a contig of 78 BACs and to completely annotate a 1.6 Mb region spanning the bovine FHIT gene. The findings confirmed the very high level of conservation between human and bovine genomes and the importance of comparative mapping to speed the annotation process of the recently sequenced bovine genome. The detailed knowledge of the genomic FHIT region will allow to study the role of FHIT in bovine cancerogenesis, especially of vesical papillomavirus-associated cancers of

  12. Tourism potential value assessment model for rural-mountain and boundary contact areas. Case study: Cluj County, the district of Ciceu and the balneal area of Bacău County (Romania

    Directory of Open Access Journals (Sweden)

    Bianca Sorina RĂCĂŞAN

    2016-11-01

    Full Text Available This paper aimed to develop a specific assessment method focused on the tourism potential of the rural-mountain and boundary contact areas. Once elaborated, the model was employed within three appropriate territories of Cluj, Bistrița and Bacău counties (Romania, who’s investigated administrative units, were repeatedly ranked into hierarchical order according to the different tourist categories, invested with numerical values. In order to reach its goals, several objectives were assigned, from awarding the components of the primary and secondary tourism supply certain scores, proposing scales and calculating values, to comparing the results and identifying best rated tourism potential categories, units and areas. With respect to the research methodology, the most commonly used methods dealt with observation, analysis and synthesis along with comparison, cartographical, statistical and mathematical techniques. Therefore, main results regarded both proposal and testing the evaluation model, highlighting values and ranging territorial units in concordance with the tourist attractiveness power.

  13. Genome-wide comparison of paired fresh frozen and formalin-fixed paraffin-embedded gliomas by custom BAC and oligonucleotide array comparative genomic hybridization: facilitating analysis of archival gliomas

    Science.gov (United States)

    Mohapatra, Gayatry; Engler, David A.; Starbuck, Kristen D.; Kim, James C.; Bernay, Derek C.; Scangas, George A.; Rousseau, Audrey; Batchelor, Tracy T.; Betensky, Rebecca A.; Louis, David N.

    2010-01-01

    Molecular genetic analysis of cancer is rapidly evolving as a result of improvement in genomic technologies and the growing applicability of such analyses to clinical oncology. Array based comparative genomic hybridization (aCGH) is a powerful tool for detecting DNA copy number alterations (CNA), particularly in solid tumors, and has been applied to the study of malignant gliomas. In the clinical setting, however, gliomas are often sampled by small biopsies and thus formalin-fixed paraffin-embedded (FFPE) blocks are often the only tissue available for genetic analysis, especially for rare types of gliomas. Moreover, the biological basis for the marked intratumoral heterogeneity in gliomas is most readily addressed in FFPE material. Therefore, for gliomas, the ability to use DNA from FFPE tissue is essential for both clinical and research applications. In this study, we have constructed a custom bacterial artificial chromosome (BAC) array and show excellent sensitivity and specificity for detecting CNAs in a panel of paired frozen and FFPE glioma samples. Our study demonstrates a high concordance rate between CNAs detected in FFPE compared to frozen DNA. We have also developed a method of labeling DNA from FFPE tissue that allows efficient hybridization to oligonucleotide arrays. This labeling technique was applied to a panel of biphasic anaplastic oligoastrocytomas (AOA) to identify genetic changes unique to each component. Together, results from these studies suggest that BAC and oligonucleotide aCGH are sensitive tools for detecting CNAs in FFPE DNA, and can enable genome-wide analysis of rare, small and/or histologically heterogeneous gliomas. PMID:21080181

  14. Seasonal patterns of carbon allocation to respiratory pools in 60-yr-old deciduous (Fagus sylvatica) and evergreen (Picea abies) trees assessed via whole-tree stable carbon isotope labeling.

    Science.gov (United States)

    Kuptz, Daniel; Fleischmann, Frank; Matyssek, Rainer; Grams, Thorsten E E

    2011-07-01

    • The CO(2) efflux of adult trees is supplied by recent photosynthates and carbon (C) stores. The extent to which these C pools contribute to growth and maintenance respiration (R(G) and R(M), respectively) remains obscure. • Recent photosynthates of adult beech (Fagus sylvatica) and spruce (Picea abies) trees were labeled by exposing whole-tree canopies to (13) C-depleted CO(2). Label was applied three times during the year (in spring, early summer and late summer) and changes in the stable C isotope composition (δ(13) C) of trunk and coarse-root CO(2) efflux were quantified. • Seasonal patterns in C translocation rate (CTR) and fractional contribution of label to CO(2) efflux (F(Label-Max)) were found. CTR was fastest during early summer. In beech, F(Label-Max) was lowest in spring and peaked in trunks during late summer (0.6 ± 0.1, mean ± SE), whereas no trend was observed in coarse roots. No seasonal dynamics in F(Label-Max) were found in spruce. • During spring, the R(G) of beech trunks was largely supplied by C stores. Recent photosynthates supplied growth in early summer and refilled C stores in late summer. In spruce, CO(2) efflux was constantly supplied by a mixture of stored (c. 75%) and recent (c. 25%) C. The hypothesis that R(G) is exclusively supplied by recent photosynthates was rejected for both species. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

  15. Expression of a gymnosperm PIN homologous gene correlates with auxin immunolocalization pattern at cotyledon formation and in demarcation of the procambium during Picea abies somatic embryo development and in seedling tissues.

    Science.gov (United States)

    Palovaara, Joakim; Hallberg, Henrik; Stasolla, Claudio; Luit, Bert; Hakman, Inger

    2010-04-01

    In seed plants, the body organization is established during embryogenesis and is uniform across gymnosperms and angiosperms, despite differences during early embryogeny. Evidence from angiosperms implicates the plant hormone auxin and its polar transport, mainly established by the PIN family of auxin efflux transporters, in the patterning of embryos. Here, PaPIN1 from Norway spruce (Picea abies [L.] Karst.), a gene widely expressed in conifer tissues and organs, was characterized and its expression and localization patterns were determined with reverse transcription polymerase chain reaction and in situ hybridization during somatic embryo development and in seedlings. PaPIN1 shares the predicted structure of other PIN proteins, but its central hydrophilic loop is longer than most PINs. In phylogenetic analyses, PaPIN1 clusters with Arabidopsis thaliana (L.) Heynh. PIN3, PIN4 and PIN7, but its expression pattern also suggests similarity to PIN1. The PaPIN1 expression signal was high in the protoderm of pre-cotyledonary embryos, but not if embryos were pre-treated with the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA). This, together with a high auxin immunolocalization signal in this cell layer, suggests a role of PaPIN1 during cotyledon formation. At later stages, high PaPIN1 expression was observed in differentiating procambium, running from the tip of incipient cotyledons down through the embryo axis and to the root apical meristem (RAM), although the mode of RAM specification in conifer embryos differs from that of most angiosperms. Also, the PaPIN1 in situ signal was high in seedling root tips including root cap columella cells. The results thus suggest that PaPIN1 provides an ancient function associated with auxin transport and embryo pattern formation prior to the separation of angiosperms and gymnosperms, in spite of some morphological differences.

  16. The mobility of nitrogen across tree-rings of Norway spruce (Picea abies L.) and the effect of extraction method on tree-ring δ¹⁵N and δ¹³C values.

    Science.gov (United States)

    Tomlinson, G; Siegwolf, R T W; Buchmann, N; Schleppi, P; Waldner, P; Weber, P

    2014-06-15

    The use of stable nitrogen (N) isotope ratios (δ(15)N values) in dendroecological studies is often preceded by an extraction procedure using organic solvents to remove mobile N compounds from tree-rings. Although these mobile N compounds may be capable of distorting potential environmental signals in the tree-ring δ(15)N values, recent investigations question the necessity of such an extraction. We used an on-going experiment with simulated elevated N deposition previously labelled with (15)N, in conjunction with control trees, to investigate the necessity of extracting mobile N compounds (using a rapid extraction procedure) for tree-ring δ(15)N and δ(13)C studies, as well as N and C concentration analyses. In addition, we examined the magnitude of radial redistribution of N across tree-rings of Norway spruce (Picea abies). The (15)N label, applied in 1995/96, was found in tree-rings as far back as 1951, although the increased N availability did not cause any significant relative increase in tree growth. The rapid extraction procedure had no significant effect on tree-ring δ(15)N or δ(13)C values in either labelled or control trees, or on N concentration. The C concentrations, however, were significantly higher after extraction in control samples, with the opposite effect observed in labelled samples. Our results indicate that the extraction of mobile N compounds through the rapid extraction procedure is not necessary prior to the determination of Norway spruce δ(15)N or δ(13)C values in dendrochemical studies. δ(15)N values, however, must be interpreted with great care, particularly when used as a proxy for the N status of trees, due to the very high mobility of N within the tree stem sapwood of Norway spruce over several decades. Copyright © 2014 John Wiley & Sons, Ltd.

  17. Determination of the Effects of Medium Composition on the Monochloramine Disinfection Kinetics of Nitrosomonas europaea by the Propidium Monoazide Quantitative PCR and Live/Dead BacLight Methods ▿

    Science.gov (United States)

    Wahman, David G.; Schrantz, Karen A.; Pressman, Jonathan G.

    2010-01-01

    Various medium compositions (phosphate, 1 to 50 mM; ionic strength, 2.8 to 150 meq/liter) significantly affected Nitrosomonas europaea monochloramine disinfection kinetics, as determined by the Live/Dead BacLight (LD) and propidium monoazide quantitative PCR (PMA-qPCR) methods (lag coefficient, 37 to 490 [LD] and 91 to 490 [PMA-qPCR] mg·min/liter; Chick-Watson rate constant, 4.0 × 10−3 to 9.3 × 10−3 [LD] and 1.6 × 10−3 to 9.6 × 10−3 [PMA-qPCR] liter/mg·min). Two competing effects may account for the variation in disinfection kinetic parameters: (i) increasing kinetics (disinfection rate constant [k] increased, lag coefficient [b] decreased) with increasing phosphate concentration and (ii) decreasing kinetics (k decreased, b increased) with increasing ionic strength. The results support development of a standard medium for evaluating disinfection kinetics in drinking water. PMID:20952645

  18. Investigations on bark extracts of Picea abies

    Energy Technology Data Exchange (ETDEWEB)

    Weissmann, G

    1981-01-01

    Successive extraction of the bark with solvents of increasing polarity yielded about 60% of soluble material. The alcohol and water extracts contained principally simple polyphenols and their glycosides, tannins, mono-and disaccharides, soluble hemicelluloses and pectins. Hot water extracts are suitable for production of adhesives by reaction with formaldehyde, but their polyphenol content is only 50%. The polyphenols and their glycosides, and glucosides of hydroxystilbenes, were investigated in detail.

  19. CRISPR/Cas9 and piggyBac-mediated footprint-free LRRK2-G2019S knock-in reveals neuronal complexity phenotypes and α-Synuclein modulation in dopaminergic neurons.

    Science.gov (United States)

    Qing, Xiaobing; Walter, Jonas; Jarazo, Javier; Arias-Fuenzalida, Jonathan; Hillje, Anna-Lena; Schwamborn, Jens C

    2017-10-01

    The p.G2019S mutation of the leucine-rich repeat kinase 2 (LRRK2) has been identified as the most prevalent genetic cause of familial and sporadic Parkinson's disease (PD). The Cre-LoxP recombination system has been used to correct the LRRK2-G2019S mutation in patient derived human induced pluripotent stem cells (hiPSCs) in order to generate isogenic controls. However, the remaining LoxP site can influence gene expression. In this study, we report the generation of a footprint-free LRRK2-G2019S isogenic hiPS cell line edited with the CRISPR/Cas9 and piggyBac technologies. We observed that the percentage of Tyrosine Hydroxylase (TH) positive neurons with a total neurite length of >2000μm was significantly reduced in LRRK2-G2019S dopaminergic (DA) neurons. The average branch number in LRRK2-G2019S DA neurons was also decreased. In addition, we have shown that in vitro TH positive neurons with a total neurite length of >2000μm were positive for Serine 129 phosphorylated (S129P) alpha-Synuclein (αS) and we hypothesize that S129P-αS plays a role in the maintenance or formation of long neurites. In summary, our footprint-free LRRK2-G2019S isogenic cell lines allow standardized, genetic background independent, in vitro PD modeling and provide new insights into the role of LRRK2-G2019S and S129P-αS in the pathogenesis of PD. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Symbiotic and nonsymbiotic hemoglobin genes of Casuarina glauca

    DEFF Research Database (Denmark)

    Jacobsen-Lyon, K; Jensen, Erik Østergaard; Jørgensen, Jan-Elo

    1995-01-01

    Frankia. Both the nonsymbiotic and symbiotic genes retained their specific patterns of expression when introduced into the legume Lotus corniculatus. We interpret this finding to mean that the controls of expression of the symbiotic gene in Casuarina must be similar to the controls of expression...... of the leghemoglobin genes that operate in nodules formed during the interaction between rhizobia and legumes. Deletion analyses of the promoters of the Casuarina symbiotic genes delineated a region that contains nodulin motifs identified in legumes; this region is critical for the controlled expression...... of the Casuarina gene. The finding that the nonsymbiotic Casuarina gene is also correctly expressed in L. corniculatus suggests to us that a comparable non-symbiotic hemoglobin gene will be found in legume species. Udgivelsesdato: 1995-Feb...

  1. Efektivita množení okrasných dřevin dřevitými řízky ve vztahu k termínu množení

    OpenAIRE

    Korbáš, Dalibor

    2016-01-01

    The object of work was experimental prove of term-impact of taking cuttings for reproduction with woody cuttings by selected cultivars and determinating the most appropriate term for cutting offtake. The experiment was established in the company Pasič LTD. in Dolní Životice on the 9th of January 2015 and ended on the 27th of August 2015. For the experiment were chosen difficult rooted cultivars of conifers: Picea abies 'Nidiformis', Picea glauca 'Conica', Taxus baccata 'Fastigiata Aurea...

  2. Direct identification of bacteria from BacT/ALERT anaerobic positive blood cultures by MALDI-TOF MS: MALDI Sepsityper kit versus an in-house saponin method for bacterial extraction.

    Science.gov (United States)

    Meex, Cécile; Neuville, Florence; Descy, Julie; Huynen, Pascale; Hayette, Marie-Pierre; De Mol, Patrick; Melin, Pierrette

    2012-11-01

    In cases of bacteraemia, a rapid species identification of the causal agent directly from positive blood culture broths could assist clinicians in the timely targeting of empirical antimicrobial therapy. For this purpose, we evaluated the direct identification of micro-organisms from BacT/ALERT (bioMérieux) anaerobic positive blood cultures without charcoal using the Microflex matrix-assisted laser desorption/ionization (MALDI) time of flight MS (Bruker), after bacterial extraction by using two different methods: the MALDI Sepsityper kit (Bruker) and an in-house saponin lysis method. Bruker's recommended criteria for identification were expanded in this study, with acceptance of the species identification when the first three results with the best matches with the MALDI Biotyper database were identical, whatever the scores were. In total, 107 monobacterial cultures and six polymicrobial cultures from 77 different patients were included in this study. Among monomicrobial cultures, we identified up to the species level 67 and 66 % of bacteria with the MALDI Sepsityper kit and the saponin method, respectively. There was no significant difference between the two extraction methods. The direct species identification was particularly inconclusive for Gram-positive bacteria, as only 58 and 52 % of them were identified to the species level with the MALDI Sepsityper kit and the saponin method, respectively. Results for Gram-negative bacilli were better, with 82.5 and 90 % of correct identification to the species level with the MALDI Sepsityper kit and the saponin method, respectively. No misidentifications were given by the direct procedures when compared with identifications provided by the conventional method. Concerning the six polymicrobial blood cultures, whatever the extraction method used, a correct direct identification was only provided for one of the isolated bacteria on solid medium in all cases. The analysis of the time-to-result demonstrated a reduction

  3. Generation of BAC Transgenic Epithelial Organoids

    NARCIS (Netherlands)

    Schwank, G.; Andersson-Rolf, A.; Koo, B.K.; Sasaki, N.; Clevers, H.

    2013-01-01

    Under previously developed culture conditions, mouse and human intestinal epithelia can be cultured and expanded over long periods. These so-called organoids recapitulate the three-dimensional architecture of the gut epithelium, and consist of all major intestinal cell types. One key advantage of

  4. Allometry of Sapwood Depth in Five Boreal Trees

    Directory of Open Access Journals (Sweden)

    M. Rebeca Quiñonez-Piñón

    2017-11-01

    Full Text Available This paper analyzes sapwood variability and allometry within individuals of Populus tremuloides, Pinus contorta, Pinus banksiana, Picea mariana, and Picea glauca. Outside bark diameter at breast height (DBH and sapwood depth (sd in four cardinal directions were measured in individuals in stands in Alberta and Saskatchewan, Canada. The microscopical analysis of wood anatomy was used to measure sd, and the error associated with the measures was observed. Sapwood allometry analyses examined the influence of DBH on sd and on sapwood area (SA. All species were observed to have varying sapwood depths around the trunk with statistical analyses showing that Pinus banksiana has a well defined preference to grow thicker in the North-East side. The largest sd values were observed for the Populus tremuloides set. Unlike Populus tremuloides and Picea glauca, for the species Pinus contorta, Pinus banksiana, and Picea mariana, incremental growth in DBH does not directly drive sapwood growth in any direction. For these three species, SA increases only because of increases in DBH as sd remains nearly constant. These results show that sapwood depth and sapwood area seem to behave differently in each studied species and are not always proportional to the tree size as is normally assumed.

  5. The timing and nature of Late Quaternary vegetation changes in the northern Great Plains, USA and Canada: a re-assessment of the spruce phase

    Science.gov (United States)

    Yansa, Catherine H.

    2006-02-01

    This paper revises the chronology for the northward migration of Picea glauca (white spruce) across the northern Great Plains, following the recession of the Laurentide Ice Sheet, and reinterprets the species composition and structure of the late-glacial vegetation on the basis of pollen and plant-macrofossil analysis. The timing of spruce migration is based on 26 14C ages obtained from Picea macrofossils. The date for the appearance of white spruce in southern South Dakota, USA, remains unchanged, 12,600 14C yr BP (ca 15,000 cal yr BP), but its arrival in southern Saskatchewan, Canada, by 10,300 14C yr BP (ca 12,100 cal yr BP) is about 1500 years later than previously estimated based on an organic sediment date. Picea glauca thus migrated northwards at an average rate of 0.38 km/ 14C year (0.30 km/calendar year), significantly slower than the previously published rate of 2 km/ 14C year. White spruce trees probably inhabited lake shorelines, whereas prairie, parkland, and boreal plants occupied both lowlands and uplands, forming an open white spruce parkland. This interpretation differs from a previous reconstruction of a boreal-type spruce forest and thus offers another paleoclimatic interpretation. Precipitation was probably low and summer temperatures relatively mild, averaging about 19 °C.

  6. Warming and neighbor removal affect white spruce seedling growth differently above and below treeline.

    Science.gov (United States)

    Okano, Kyoko; Bret-Harte, M Syndonia

    2015-01-01

    Climate change is expected to be pronounced towards higher latitudes and altitudes. Warming triggers treeline and vegetation shifts, which may aggravate interspecific competition and affect biodiversity. This research tested the effects of a warming climate, habitat type, and neighboring plant competition on the establishment and growth of white spruce (Picea glauca (Moench) Voss) seedlings in a subarctic mountain region. P. glauca seedlings were planted in June 2010 under 4 different treatments (high/control temperatures, with/without competition) in 3 habitats (alpine ridge above treeline/tundra near treeline /forest below treeline habitats). After two growing seasons in 2011, growth, photosynthesis and foliar C and N data were obtained from a total of 156, one-and-a-half year old seedlings that had survived. Elevated temperatures increased growth and photosynthetic rates above and near treeline, but decreased them below treeline. Competition was increased by elevated temperatures in all habitat types. Our results suggest that increasing temperatures will have positive effects on the growth of P. glauca seedlings at the locations where P. glauca is expected to expand its habitat, but increasing temperatures may have negative effects on seedlings growing in mature forests. Due to interspecific competition, possibly belowground competition, the upslope expansion of treelines may not be as fast in the future as it was the last fifty years.

  7. Volatile monoterpenes in a stand of Picea abies (L.) Karst

    International Nuclear Information System (INIS)

    Schuermann, W.; Eichstaedter, G.; Steinbrecher, R.; Schoenwitz, R.; Gabriel, R.; Karbach, I.; Zufall, N.; Ziegler, H.

    1992-01-01

    Natural emissions of terpene and their fate in the atmosphere were investigated in a forest stand of the Bayerischer Wald mountains. Terpenes are volatile hydrocarbons whose relevance for air chemistry has increased considerably in the past few years. The investigation compared different sources of terpene in order to determine their strength as a function of meteorological, air-chemical and biological parameters. (orig./BBR) [de

  8. Enzyme activities of fungi associated with Picea abies needles

    Czech Academy of Sciences Publication Activity Database

    Žifčáková, Lucia; Dobiášová, Petra; Kolářová, Zuzana; Koukol, O.; Baldrian, Petr

    2011-01-01

    Roč. 4, č. 6 (2011), s. 427-436 ISSN 1754-5048 R&D Projects: GA MŠk LC06066; GA ČR GA526/08/0751 Institutional research plan: CEZ:AV0Z50200510 Keywords : Ascomycetes * Decomposition * Basidiomycetes Subject RIV: EE - Microbiology, Virology Impact factor: 2.507, year: 2011

  9. Aerial photographs with infra-red film to survey injury to vegetation in Styria

    Energy Technology Data Exchange (ETDEWEB)

    Pollanschuetz, J

    1973-01-01

    This paper describes a successful operation in which injuries to Norway Spruce, chiefly caused by SO/sub 2/ pollution from smelters, was mapped from the air. Four zones varying in the severity of injury could be distinguished. They were characterized chiefly by differences in the number and age of needles present on the trees. The fourth zone contained only scattered, severely injured, but apparently highly resistant individual trees of Spruce. The photographs, however, did not reveal injuries that were not already visible to the naked eye. The relative resistance to air pollution of Blaufichte (Picea glauca) was noticed incidentally. 6 references.

  10. Effects on the forest of sulfur dioxide from a sulfur fire near Edson, Alberta

    Energy Technology Data Exchange (ETDEWEB)

    Hocking, D

    1975-01-01

    Sulfur was burnt in a sanitary landfill during August 9 and 10, 1974. Resulting sulfur dioxide impinged on the surrounding mixed forest for 29 h. About 4 ha of forest displayed visible injury symptoms of varying intensity soon after. However, only .4 ha remained permanently injured the next season. Here, white spruce (Picea glauca (Moench) Voss) and scattered individuals of balsam poplar (Populus balsamifera L.), alder (Alnus tenuifolia Nutt.), and trembling aspen (Populus tremuloides Michx.) were killed. This report describes the extent of injury, relative sensitivities of affected plant species, and recovery in the spring in 1975.

  11. Annotation and BAC/PAC localization of nonredundant ESTs from ...

    Indian Academy of Sciences (India)

    Unknown

    2002-04-25

    Apr 25, 2002 ... School of Life Sciences, University of Hyderabad, Hyderabad 500 046, India. Abstract ... Putative functions were assigned at a stringency E value of 10–6 in ... drought and salinity tolerance governed by multiple genes, as ...

  12. 110 A CRITICAL DISCOURSE ANALYSIS OF REV. FR. PROF. BAC ...

    African Journals Online (AJOL)

    seeing the text as the product of a process of production, and as a resource in the ... Mind control is, therefore, a function of texts and talks, and does not always mean ... does not mean critique in the negative sense of it, but fostering in-depth ... refers to the social factors that lead to the production of a text and the effect of ...

  13. Exploiting repetitive sequences and BAC clones in Festuca pratensis karyotyping

    Czech Academy of Sciences Publication Activity Database

    Majka, J.; Ksiazczyk, T.; Kiełbowicz-Matuk, A.; Kopecký, David; Kosmala, A.

    2017-01-01

    Roč. 12, č. 6 (2017), č. článku e0179043. E-ISSN 1932-6203 Institutional support: RVO:61389030 Keywords : IN-SITU HYBRIDIZATION * RIBOSOMAL-RNA GENES * LOLIUM-MULTIFLORUM Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Plant sciences, botany Impact factor: 2.806, year: 2016

  14. Fontes hidrominerais do Brasil: componentes biologicamente ativos (BAC) naturais

    OpenAIRE

    Lazzerini, Fábio Tadeu [UNESP

    2013-01-01

    Os ambientes e recursos naturais de fontes hidrominerais devem fazer parte da governança e planejamento estratégico na saúde pública, meio ambiente, bem estar, turismo e mineração. Na atual política nacional observada em: termalismo social/crenoterapia como prática integrativa complementar da medicina (PIC) através da Portaria Ministério da Saúde 971/2006; na exploração e aplicação das águas minerais prevista pelas Portarias Ministério das Minas e Energia 337/2002 e 127/2011; na implementação...

  15. Waarnemingen omtrent den bacteriophaag bij Bac. danicus en B. radicicola

    NARCIS (Netherlands)

    Grijns, A.

    1926-01-01

    Bacteriophages of Bacillus danicus and Bacterium radicicola were isolated from soil and root nodules. An extensive study was made on the effect of heat in the inactivation of bacteriophage of B. danicus. A chemical analysis of these cultures of B. danicus with or without bacteriophages showed a

  16. Rhipicephalus microplus strain Deutsch, 10 BAC clone sequences

    Science.gov (United States)

    The cattle tick, Rhipicephalus (Boophilus) microplus, has a genome over 2.4 times the size of the human genome, and with over 70% of repetitive DNA, this genome would prove very costly to sequence at today's prices and difficult to assemble and analyze. We used labeled DNA probes from the coding reg...

  17. Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens

    KAUST Repository

    Ali, Shawkat; Bakkeren, Guus

    2014-01-01

    The genetic transformation of certain organisms, required for gene function analysis or complementation, is often not very efficient, especially when dealing with large gene constructs or genomic fragments. We have adapted the natural DNA transfer

  18. Optimization of enzymatic cartilage hydrolysis from Prionace glauca wastes for the production of chondroitin sulphate

    OpenAIRE

    Vázquez, José Antonio; Blanco, M.; Noriega, Diana; Fraguas, Javier; Pastrana, Lorenzo; González Sotelo, Carmen; Pérez Martín, Ricardo Isaac

    2014-01-01

    Comunicación presentada en el Total Food Conference 2014 Science and technology for the economic and sustainable exploitation of agri-food chain wastes and co-products, Norwich, Uk, 11-13 November (2014)

  19. Photoenzymatic repair of ultraviolet-irradiated DNA in the cells of a shark, Prionace glauca

    International Nuclear Information System (INIS)

    Woodhead, A.D.; Achey, P.; Setlow, R.B.; Grist, E.

    1978-01-01

    Cells from several tissues of the blue shark have been tested for the presence of photoreactivating enzyme using a sensitive assay requiring only 250 ng of DNA. Photoactivating enzyme was found to be present in thyroid, liver, brain and kidney cells. There has been much debate concerning the significance of the reported distribution of photoreactivating enzyme in animal tissues, but the information available is too limited for generalizations to be made. It is suggested that the present assay might allow rapid and easy survey of a wide spectrum of organisms. (author)

  20. The effect of intermittent dosing of Nicotiana glauca on teratogenesis in goats

    Science.gov (United States)

    Sustained inhibition of fetal movement in livestock species, induced by several poisonous plants, can result in numerous skeletal-contracture malformations. Lupines are responsible for a condition in cattle referred to as “crooked calf syndrome” that occurs when pregnant cattle graze teratogenic lup...