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  • 1
    ISSN: 1572-8773
    Keywords: NMR and EPR spectroscopy ; speciation ; transformation ; vanadate ; vanadyl ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Three strains of Saccharomyces cerevisiae, SC-1, DBVPG 6173 and DBVPG 6037, were studied for vanadate resistance in complex Sabouraud medium since they did not thrive in different minimal media (yeast nitrogen base with and without amino acids). The strain SC-1 was resistant up to 16 mm of vanadate, whereas the strains DBVPG 6173 and DBVPG 6037 were inhibited by 8 mm and 4 mm vanadate, respectively. The vanadate resistance in strain SC-1 was constitutive and due to the reduction of this oxyanion to vanadyl, which was detected by EPR spectroscopy and visible spectroscopy. The transformation of vanadate to vanadyl took place during the exponential growth phase; 10 mm of vanadate was reduced to vanadyl outside the cells since the oxyanion was not detected in the cell biomass and only a negligible concentration of vanadyl (25 nmoles mg cells dry weight) was found in the biomass. The other two vanadate-sensitive yeast strains only accumulated vanadate and did not reduce the oxyanion to vanadyl.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Catalysis letters 49 (1997), S. 121-128 
    ISSN: 1572-879X
    Keywords: cresols ; transformation ; isomerization ; disproportionation ; zeolites HY
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The transformation of m- and o-cresol has been investigated on a series of HY zeolites with Si/Al ratio in the range of 4.5 to 55, at 380°C and atmospheric pressure. It was found that the zeolite activity increased but stability decreased with NAl for both m- and o-cresol. For Si/Al = 4.5 the zeolite activity was similar for both reactants, but as Si/Al ratio increased, o-cresol became relatively more reactive than m-cresol, and for all Si/Al ratios the catalysts were more stable in the m-cresol transformation. Both m- and o-cresol were converted mainly through two parallel reactions - isomerization and disproportionation. The disproportionation reaction was predominant, and more important in the case of o-cresol. The isomerization/disproportionation selectivity increased for m-cresol and decreased for o-cresol with NAl. Catalyst deactivation favoured disproportionation in the case of o-cresol and isomerization in the case of m-cresol, except for o-cresol conversion on HY(4.5), where both reactions were equally affected. Isomerization selectivity for m-cresol (p/o) was found to be higher than the equilibrium value and to increase with catalyst deactivation; while for o-cresol, the isomerization ratio (m/p) did not change with time on stream for HY(4.5) and HY(16.6) and decreased for HY(55). Disproportionation selectivity (ph/∑x) was equal to unity independently of Si/Al ratio or catalyst deactivation.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    International journal of value-based management 10 (1997), S. 9-29 
    ISSN: 1572-8528
    Keywords: business ethics ; Jewish ; religious ; community ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Economics
    Notes: Abstract This paper examines the method of Jewish business ethics. MichaelWalzer, in his work, Interpretation and Social Criticism (1987), suggeststhree common and important approaches to moral philosophy. He labels thesethe path of discovery, the path of invention, and the path ofinterpretation. The first part of this paper argues that Jewish businessethics is best thought of in terms of interpretation. Without question, thereligious ethicist immediately recognizes Walzer‘s metaphor of the moralworld as a ’home occupied by a single family over many generations... ‘as his own. Ethical arguments from a Jewish perspective must of necessityhave a ’lived-in quality‘ and always make reference to and are based on the’memory-laden objects and artifacts.‘ The second part of the paper exploressome of the implications of Jewish business ethics as interpretation.
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  • 4
    ISSN: 1432-0789
    Keywords: Cyanobacteria ; Soil inoculation N ; transformation ; Inorganic N ; Easily oxidizable N ; Hydrolysable N ; Non-hydrolysable N ; Wetland rice Farmyard manure ; Oryza sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A field experiment was conducted with wetland rice (Oryza sativa cv. IR-36) in a sandy clay loam soil (Entisol) to study the effect of inoculation with a soil-based mixed culture of four diazotrophic cyanobacteria,Aulosira fertilissima, Nostoc muscorum, N. commune andAnabaena spp., on the N-flux in inorganic NH4 ++NO3 −+ NO2 −), easily oxidizable, hydrolysable and non-hydrolysable forms of N in soil during vegetative growth periods of the crop. Effects on grain and straw yield and N uptake by the crop were estimated. The effects of applying urea N and N as organic sources, viz.Sesbania aculeata, Neem (Azardirachta indica) cake and FYM, each at the rate of 40 kg N ha−1, to the soil were also evaluated. Inoculation significantly increased the release of inorganic N, evidenced by its increased concentrations either in soil or in soil solution. However, such increases rarely exceeded even 4% of total N gained in different froms in the soil system by inoculation during the vegetative growth stages of the rice plant, when the nutritional requirement of the plants is at a maximum. Most of the N2 fixed by cyanobacteria remained in the soil as the hydrolysable form (about 85%) during this period. Inoculation caused an insignificant increase in grain (8%) and straw (11%) yield, which was, however, accompanied by a significant increase in N uptake by the grain (30%) and an increase in total uptake of 15.3 kg N ha 1. Such beneficial effects of inoculation varied in magnitude with the application of organic sources, with farmyard manure (FYM) being the most effective. Application of urea N, on the other hand, markedly reduced such an effect.
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  • 5
    ISSN: 1572-9818
    Keywords: Agrobacterium ; Populus ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In recent years, Populus species have acquired an important place in basic and applied research of woody plants. The practical role of Populus species in world forestry and their importance to research as a woody-plant model have led to increasing interest in tissue-culture and molecular techniques, as well as the development of transformation procedures for this genus. A simple technical procedure is described here step-by-step, for the first time, as a routine method for transforming Populus tremula using a disarmed Agrobacterium tumefaciens hypervirulent strain. The procedure begins with the inoculation of stem explants with bacterial suspension, followed by a short period of co-cultivation on a highly regenerative medium. Transformed shoots are selected on regeneration medium containing antibiotics and the presence of the inserted target genes is checked using a rapid and efficient PCR test. Selected shoots are transferred to a rooting medium, under the same selection pressure, and propagated via stem cuttings. Selected plants can be hardened and transferred to the green-house within 4 months of inoculation. The method has proven efficient for several gene constructs, selection on Kan or Hyg, and three different Agrobacterium strains.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology reporter 15 (1997), S. 209-215 
    ISSN: 1572-9818
    Keywords: selectable marker ; cereals ; leaf assay ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A very simple leaf assay is described that rapidly and reliably identifies transgenic plants expressing the hygromycin resistance gene, hph or the phosphinothricin resistance gene, bar. Leaf tips were cut from plants propagated either in the glasshouse or in tissue culture and the cut surface embedded in solid medium containing the appropriate selective agent. Non-transgenic barley or rice leaf tips had noticeable symptoms of either bleaching or necrosis after three days on the medium and were completely bleached or necrotic after one week. Transgenic leaf tips remained green and healthy over this period. This gave unambiguous discrimination between transgenic and non-transgenic plants. The leaf assay was also effective for dicot plants tested (tobacco and peas).
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  • 7
    ISSN: 1572-9818
    Keywords: Junction sequence ; transformation ; Agrobacterium tumefaciens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genomic integration of transferred T-DNA is traditionally analyzed by Southern hybridization; however, these analyses often do not provide sufficient information pertaining to the transformation event. Analysis of the junction sequences spanning the region between the T-DNA borders and plant genomic DNA, give a clear demonstration of genomic integration. The procedures available for border junction analysis can be problematic, therefore a simplified method was developed for plants transformed by Agrobacterium tumefaciens harboring the binary vector with pBI121 backbone.
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  • 8
    ISSN: 1573-4919
    Keywords: endothelin ; fibroblasts ; receptors ; transformation ; oncogene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Endothelin-1 (ET-1), a peptide isolated from the culture medium of endothelial cells, mediates a variety of physiological and pathological responses including mitogenesis. We have compared the expression of ET receptors in untransformed versus ras-transformed NIH-3T3 murine fibroblasts and in untransformed versus SV40-transformed Wl38 (VA13) human fibroblasts by ligand binding and Northern analysis. NIH-3T3 and Wl38 cells displayed high affinity (200 and 220 pM) and high density (23,000 sites/cell and 14,000 sites/cell for NIH-3T3 and Wl38 cells, respectively) ET receptors. Competition binding experiments using subtype-selective ligands identified these receptors as the ETA subtype. Addition of ET-1 to the cells produced a concentration-dependent increase in intracellular calcium release. Both ras-transformed NIH-3T3 cells and SV40-transformed Wl38 cells (VA13) completely lacked [125I]ET-1 binding and failed to release calcium when exposed to ET-1. Northern analysis of the polyadenylat ed RNA (polyA RNA) isolated from untransformed and transformed cells revealed that the steady-state level of ETA receptor RNA was 90-95% less in transformed cells compared to untransformed cells. Thus, the loss of ET receptors as well as the receptor-mediated responses in transformed cells can be explained by down-regulation of ET receptor mRNA.
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  • 9
    ISSN: 1573-5028
    Keywords: CaMV 35S promoter ; Catharanthus roseus ; FMV 34S promoter ; particle bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Activities of several CaMV 35S and FMV 34S promoter derivatives fused to the gusA reporter gene were compared in suspension-cultured Catharanthus roseus cells that were transiently and stably transformed using particle bombardment. Our data demonstrate that the 35S and a deletion derivative of the 34S promoter combined with particle bombardment form useful tools for genetic engineering of C. roseus cells. Our results disagree on several points with activities of 35S and 34S promoter derivatives reported for tobacco, indicating that absolute and relative promoter activities can differ between plant species.
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  • 10
    ISSN: 1573-9368
    Keywords: Ac ; Agrobacterium ; aspen ; Populus ; transformation ; transposition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aspen (Populus tremula) and hybrid aspen (P. tremula × P. tremuloides) were transformed with different gene constructs using two types of promoter. The aim was to determine the influence of the reporter gene rolC, controlled by promoters of viral or plant origin, on genetic and morphologic expression of different transgenic aspen clones. An improved transformation method using leaf discs was developed, by which putative transgenic plantlets were regenerated at high efficiencies (up to 34%) on kanamycin-containing medium. Transgenic aspen carrying the rolC gene from Agrobacterium rhizogenes under control of the cauliflower-35S-promoter are reduced in size with smaller leaves, whereas aspen transgenic for the same rolC gene, but under control of the light inducible rbcS promoter from potato, are only slightly reduced in size compared to untransformed controls. However, all clones carrying 35S-rolC and rbcS-rolC genes revealed light-green colouration of leaves when compared to untransformed aspen. Owing to this special feature, constructs were used in which expression of the rolC gene was inhibited by insertion of a transposable element, Ac, from maize. Transgenic aspen transformed with the 35S-Ac-rolC and rbcS-Ac-rolC genes were morphologically similar to untransformed aspen, but out of 54 independently regenerated 35S-Ac-rolC transgenic aspen clones, 30 clones showed light-green/dark green variegated leaves. In contrast, out of 19 independently transformed rbcS-Ac-rolC aspen clones, only two clones revealed light-green/dark green variegated leaves. The role of bacterial strains in transformation, and molecular genetics of transgenic aspen plants (including the function of the transposable element, Ac, in the aspen genome) are discussed
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  • 11
    ISSN: 1573-9368
    Keywords: peanut ; engineered virusresistance ; tomato spotted wiltvirus ; transformation ; nucleocapsid ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleocapsid gene of tomato spotted wilt virus Hawaiian L isolate in a sense orientation, and the GUS and NPTII marker genes, were introduced into peanut (Arachis hypogaea cv. New Mexico Valencia A) using Agrobacterium-mediated transformation. Modifications to a previously defined transformation protocol reduced the time required for production of transformed peanut plants. Transgenes were stably integrated into the peanut genome and transmitted to progeny. RNA expression and production of nucleocapsid protein in transgenic peanut were observed. Progeny of transgenic peanut plants expressing the nucleocapsid gene showed a 10- to 15-day delay in symptom development after mechanical inoculations with the donor isolate of tomato spotted wilt virus. All transgenic plants were protected from systemic tomato spotted wilt virus infection. Inoculated non-transformed control plants and plants transformed with a gene cassette not containing the nucleocapsid gene became systemically infected and displayed typical tomato spotted wilt virus symptoms. These results demonstrate that protection against tomato spotted wilt virus can be achieved in transgenic peanut plants by expression of the sense RNA of the tomato spotted wilt virus nucleocapsid gene
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  • 12
    ISSN: 1573-9368
    Keywords: transgenic cotton ; bialaphos ; particle bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Resistance to bialaphos, a non-selective herbicide, was intro duced into cotton through genetic engineering. A gene encoding phosphinothric in acetyltransferase (bar) from Streptomyces hygroscopicus was inserted into elite varieties of cotton through particle bombardment. Based on the marker gene, β-glucuronidase (gus) expression, a total of 18 Pima (Gossypium barbadense), 45 DP50 (G. hirsutum L.), 20 Coker 312 (G. hirsutum) and 2 El Dorado (G. hirsutum) transgenic plants were recovered. Integration of the bar gene into cotton genomic DNA was confirmed by Southern blot analysis and gene expression was confirmed by northern blot and enzyme assays. Herbicide (Basta®) tolerance up to 15 000 ppm was demonstrated in greenhouse trials. The newly introduced herbicide tolerance trait is inherited in a Mendelian fashion in the progenies of germline transformants. This study demonstrates the potential for particle bombardment to introduce commerically important genes directly into elite varieties of cotton. This mode of gene transfer can expedite the introduction of transgenic cotton products into world markets
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Transgenic research 6 (1997), S. 279-288 
    ISSN: 1573-9368
    Keywords: Agrobacterium tumefaciens ; Brassica napus ; wintercultivar ; transformation ; GUS ; hygromycin ; kanamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient protocol for Agrobacterium tumefaciens-mediated transformation of six commercial Brassica napus winter cultivars is described. Two B. napus spring cultivars were analysed for comparison. Five strains of A. tumefaciens with different combinations of nopaline and octopine chromosomal backgrounds and virulence plasmids were used for cocultivation. Selection of putative regenerated transgenic plants was performed on kanamycin- or hygromycin-containing media. The scores of transgenic plants were calculated on the basis of GUS (β-glucuronidase) activity, detected by the histochemical X-Gluc test. Target tissue derived from the cut surface of cotyledon petioles resulted in successful transformation with all the winter cultivars tested. Target tissue from hypocotyl segments resulted in a successful transformation with only one winter cultivar. The transformation rates for B. napus winter cultivars in this study were higher than in previous reports. Southern blot analysis revealed that integration of marker genes occurred in single and in multiple copies and at multiple loci in the genome. The transgenic plants all grew normally and developed fertile flowers after a vernalization period. After self-pollination, Southern blot analysis of selected GUS active F1 plants revealed that introduced marker genes were stably inherited to the next generation. These data demonstrate that morphologically normal, fertile transgenic plants of B. napus winter cultivars can be achieved with both nopaline- and octopine-derived A. tumefaciens strains. This protocol should have a broad application in improvement of Brassica napus winter cultivars by introduction of foreign genes
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  • 14
    ISSN: 1573-9368
    Keywords: transformation ; Erysiphe graminis ; powderymildew ; coleoptile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A transient assay is described that should allow evaluation of the role of host genes in disease response by enhancing or disrupting expression of those genes in specific cells and looking for effects on disease development. The assay also has the potential for assessing utility of host and non-host genes in enhancing resistance to disease in transgenic plants. Particle bombardment with a helium discharge particle gun was utilized to transiently express genes in epidermal cells of coleoptiles of barley (Hordeum vulgare). An anthocyanin reporter gene construct provided a means of identifying those cells that were transiently expressing introduced DNA. Optimal transient expression rates were achieved two days following bombardment with 1800 psi helium pressure, 1.0 μm diameter gold particles, and coleoptile pre- and post-treatment in 0.30--0.35 m mannitol/sorbitol. Under optimal conditions, at least 35 cells expressed anthocyanin per bombardment. Transiently expressing cells were inoculated with the fungal pathogen, Erysiphe graminis f. sp. hordei, and fungal development observed. Neither the bombardment procedures, the presence of nearby dead cells, nor accumulation of anthocyanin within living cells affected fungal development in living cells. Therefore, incorporation of disease-related genes onto the same plasmid as the reporter genes will allow evaluation of the role of those genes in disease development or suppression. Since particle bombardment is possible with a great range of different plant tissues, the described methodology should exhibit wide applicability for evaluating genes in diverse plant-pathogen interactions, as well as genes involved in many other biological processes
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Journal of porous materials 4 (1997), S. 165-169 
    ISSN: 1573-4854
    Keywords: ferrihydrite ; hematite ; BET ; porosity ; catalyst ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract Ferrihydrite is a poorly-ordered iron oxyhydroxide which possesses a high specific surface area (〉200 m2/g). This makes it an important constituent for surface related processes in nature and catalysis. This study focuses on the precautions that have to be taken in the sample preparation prior to BET measurements in order to avoid essential structural transformations. The experiments showed that temperatures between 90 and 120°C are necessary to remove surface contaminations and simultaneously prevent structural transformation of the ferrihydrite during the outgassing procedure. Furthermore, care has to be taken to keep the humidity low in the immediate proximity of the sample, due to the strong impact of this parameter on the structural stability of ferrihydrite.
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  • 16
    ISSN: 1572-9788
    Keywords: Bacillus thuringiensis ; Chilo suppressalis ; biolistic ; rice ; Scirpophaga incertulas ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Rice cultivars of isozyme group V include high-quality, aromatic rices that are difficult to improve by traditional methods because of the loss of quality characters upon sexual hybridization. Their low-tillering plant type predisposes them to economic loss from attack by stem borers, a group of insects to which they are susceptible. We report here the enhancement of stem borer resistance in cv. Tarom Molaii through transformation by microprojectile bombardment. Embryogenic calli derived from mature seeds were bombarded with gold particles coated with plasmid pCIB4421, carrying a synthetic truncated toxin gene based on the cryIA(b) gene from Bacillus thuringiensis, and plasmid pHygII, carrying the hygromycin phosphotransferase (hpt) selectable marker gene. Inclusion of 50 mg/l hygromycin B in culture media from bombardment through to rooting of plantlets eliminated escapes. The procedure generated three independent hpt transformants of which two also contained the cryIA(b) gene. One such line (No. 827) produced truncated (67 kDa) CryIA(b) protein equivalent to about 0.1% of total soluble protein. The cryIA(b) gene was controlled by the promoter of the maize C4 PEP carboxylase gene and was expressed in leaf blades but was not expressed to a detectable level in dehulled mature grain. Line 827 contained about 3 copies of the cryIA(b) gene which segregated as a single dominant Mendelian locus in the second (T1) and third (T2) generations and co-segregated with enhanced resistance to first-instar larvae of striped stem borer (Chilo suppressalis) and yellow stem borer (Scirpophaga incertulas). T2 line 827-6 homozygous for the cryIA(b) gene showed no dead hearts or whiteheads after infestation with stem borers, whereas T2 line 827-25 lacking the gene averaged 7 dead hearts per plant and 2.25 whiteheads per plant. These results establish that transformation of high-quality rices of group V is a feasible alternative to sexual hybridization.
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  • 17
    ISSN: 1572-9788
    Keywords: adventitious root formation ; Agrobacterium tumefaciens ; A. rhizogenes ; rootstock ; rose ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Transgenic plants of the rootstock Rosa hybrida L. cv. Moneyway were produced via a two-step procedure. First, kanamycin-resistant roots were generated on stem slices from micropropagated shoots, which were cocultivated with Agrobacterium tumefaciens containing the neomycin phosphotransferase II (NPTII) gene for conferring kanamycin resistance, together with individual ROL genes from A. rhizogenes. Root formation was quite efficient and up to two kanamycin-resistant roots per stem slice were produced. In the second step, these roots were used to regenerate transgenic plants via somatic embryogenesis. Although regeneration lasted up to 12 months, production of several transformants was successfully accomplished. Untransformed escapes were not found, indicating that the initial selection on kanamycin resistance was reliable. The presence of a combination of ROLA, B and C genes enhanced adventitious root formation on micropropagated shoots and explants of stems and leaves. It appears that the auxin sensitivity was increased to such a degree that cells were able to respond even to endogenous auxins present in shoots and leaves. Rooting experiments in greenhouse demonstrated that adventitious root formation on cuttings was improved threefold upon introduction of these ROL genes. It is concluded that a method was developed for the production of ROL gene transformed roses with improved rooting characteristics.
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  • 18
    ISSN: 1573-5028
    Keywords: competitive PCR ; flavonoid pathway ; Forsythia ; gene expression ; transformation ; woody ornamentals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression, during flower development, of the gene encoding the anthocyanin pathway key enzyme dihydroflavonol 4-reductase (DFR) was investigated in floral organs of Forsythia × intermedia cv. ‘Spring Glory’. Full-length DFR and partial chalcone synthase (CHS) cDNAs, the gene of interest and a flavonoid pathway control gene respectively, were obtained from petal RNA by reverse transcription PCR. Whereas for CHS northern blot analysis enabled the study of its expression pattern, competitive PCR assays were necessary to quantify DFR mRNA levels in wild-type plants and in petals of 2 transgenic clones containing a CaMV 35S promoter-driven DFR gene of Antirrhinum majus. Results indicated a peak of CHS and DFR transcript levels in petals at the very early stages of anthesis, and different expression patterns in anthers and sepals. In comparison to wild-type plants, transformants showed a more intense anthocyanin pigmentation of some vegetative organs, and a dramatic increase in DFR transcript concentration and enzymatic activity in petals. However, petals of transformed plants did not accumulate any anthocyanins. These results indicate that other genes and/or regulatory factors should be considered responsible for the lack of anthocyanin production in Forsythia petals.
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  • 19
    Electronic Resource
    Electronic Resource
    Springer
    Pharmaceutical research 14 (1997), S. 750-756 
    ISSN: 1573-904X
    Keywords: DNA delivery ; cells ; sonication ; cavitation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. Ultrasonic methods have considerable potential for the introduction of macromolecules into cells. In this paper we demonstrate that, under controlled conditions, application of 20kHz ultrasound to a suspension of yeast cells facilitates the delivery of plasmid DNA into these cells. Methods. Aliquots of growing yeast cells (Saccharomyces cerevisae, strain AH22) were suspended in buffer and exposed to 20kHz ultrasound from a laboratory (probe-type) sonicator in the presence of microgram quantities of plasmid DNA. Efficiency of DNA delivery was scored as the number of cells transformed. Results. Cell transformation was optimal at 30 seconds sonication using an output of 2.0 watts and resulted in a 20 fold enhancement over control values. At extended sonication times, fewer cells showed evidence of transformation because of reduced cell viability. The increased DNA uptake and the decreased cell viability were both attributable to acoustic cavitation events during sonication. The extent of acoustic cavitation was measured and it was found that there was an increase in cavitation events with increased sonication time. Cell viability was shown to be directly related to the number of cavitation events. The effects of sonication on plasmid DNA were investigated and indicated that the structural integrity of plasmid DNA was unaffected by the sonication conditions employed. Conclusions. Under controlled conditions, ultrasound is an effective means of delivering plasmid DNA into cells. The subsequent expression of DNA molecules in cells depends upon a balance between transient cell damage and cell death.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 94 (1997), S. 83-91 
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; Beta vulgaris ; 5-enolpyruvylshikimate-3-synthase ; glyphosate ; herbicide ; sugar beet ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Sugar beet (Beta vulgaris L.) lines transformed with the 5-enolpyruvylshikimate-3-phosphate synthase gene (CP4 EPSPS) from Agrobacterium sp. CP4 and a glyphosate oxidase reductase gene (GOX) also isolated from bacteria resulted in the development of lines highly tolerant to glyphosate. Glyphosate (N-phosphonomethyl-glycine) is the active ingredient in Roundup®, herbicide. The EPSPS enzyme is involved in the biosynthesis of aromatic amino acids. Glyphosate binds irreversible to the EPSPS and inhibits the pathway. GOX degrades glyphosate into non-toxic compounds. 260 independent transformants have been evaluated in greenhouse and field trials for tolerance to Roundup® in 1993 and 1994. Variation of tolerance was recorded between different transformants, ranging from complete susceptibility to full tolerance. The Agrobacterium tumefaciens mediated transformation resulted in a negative correlation between copy number of the T-DNA insert and the level of tolerance to the herbicide. Transformants which contain a single copy insert showed tolerance to higher doses of glyphosate than transformants with multiple copies. Two transgenic lines were identified that showed agronomically useful tolerance to glyphosate.
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  • 21
    ISSN: 1573-5079
    Keywords: Chlamydomonas reinhardtii ; mutagenesis ; photoinhibition ; Photosystem II ; repair cycle ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The work outlines the isolation of transformant Chlamydomonas reinhardtii cells that appear to be unable to repair Photosystem II from photoinhibitory damage. A physiological and biochemical characterization of three mutants is presented. The results show differential stability for the D1 reaction center protein in the three mutants compared to the wild type and suggest lesions that affect different aspects of the Photosystem II repair mechanism. In the ag16.2 mutant, significantly greater amounts of D1 accumulate in the thylakoid membrane than in the wild type under steady-state growth conditions, and D1 loss is significantly retarded in the presence of the protein biosynthesis inhibitor chloramphenicol. Moreover, aberrant electrophoretic mobility of D1 in the ag16.2 suggests that this protein is modified to an as yet unknown configuration. These results indicate that the biosynthesis and/or degradation of D1 is altered in this strain. A different type of mutation occurred in the kn66.7 and kn27.4 mutants of C. reinhardtii. The stability of D1 declined much faster as a function of light intensity in these mutants than in the wild type. Thereby, the threshold of photoinhibition in these mutants was significantly lower than that in the wild type. It appears that kn66.7 and kn27.4 are similar conditional mutants, with the only difference between them being the amplitude of the chloroplast response to the mutation and the differential sensitivity they display to the level of irradiance.
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  • 22
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 49 (1997), S. 71-74 
    ISSN: 1573-5044
    Keywords: cocultivation ; monocot ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Root explants from in vitro-grown plants of Anthurium andraeanum ‘Alii’ and Anthurium interspecific hybrid ‘UH1060’ readily produced multiple shoots under weak light on a modified Murashige & Skoog medium containing 2.2 μM BA. Regenerated ‘UH1060’ plants grew normally and flowered within 16 months after transfer to the greenhouse. Cocultivation of root cuttings with Agrobacterium tumefaciens LBA4404 carrying the binary vector pCa2Att resulted in kanamycin-resistant ‘Anuenue’ shoots transgenic for neo and att and recovered more than one year after culture on selection media. Transformation efficiency (number of explants with transgenic shoots per total explants) was 1.3%. Other cultivars (‘UH1003’, ‘UH1060’, ‘Rudolph’, and ‘Mauna Kea’) failed to produce shoots under the transformation condition employed.
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  • 23
    ISSN: 1573-5044
    Keywords: Agrobacterium ; coat protein ; grapevine ; hairy root ; nepovirus ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hairy root cultures of grapevine were obtained from plantlets co-inoculated by virulent Agrobacterium rhizogenes strains and disarmed A. tumefaciens strains harbouring the binary vectors pKHG4 and pKVHG 2+. These plasmids contain the nptII, hpt and gus genes and differ for the presence of the gene encoding for the grapevine chrome mosaic virus coat protein. For the cultivar ‘Gravesac’, 72% of the excised root tips initiated hairy root cultures on growth regulator-free media. According to the nature of the strains used in co-inoculation, co-transformation frequencies of the hairy root clones ranged from 4 to 16%. Co-transformed roots showed resistance to kanamycin and hygromycin but responses varied from clone to clone. Fluorometric GUS expression and GCMV coat protein production showed a large variability among hairy root clones co-transformed by pKHVG2+. Though the presence of gus, nptII and GCMV coat protein genes was checked by polymerase chain reaction and Southern blotting, it was difficult to establish a clear relationship between expression of the different transgenes. The regeneration of plants was not achieved, but the possibility to graft in vitro transgenic roots to non transformed shoot systems could permit rapid testing of the resistance induced by nepovirus coat protein in roots of cultivars that are recalcitrant to A. tumefaciens-mediated transformation.
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  • 24
    ISSN: 1573-6857
    Keywords: Anopheles albimanus ; malaria vector ; transformation ; white gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have cloned and characterized the white gene of Anopheles albimanus. Comparison of the deduced amino acid sequence of this white gene with its homologs from six species of Diptera show that the An. albimanus gene is most similar to the white gene of An. gambiae (92% identity). A spontaneous white-eyed mutant An. albimanus was caused by an approximately 10 kb insertion into a CT dinucleotide repeat region of intron 2 of the white locus. The flanks of this insertion are long (at least 400 bp), nearly perfect inverted terminal repeat sequences. This cloned white gene should be useful as a marker for germ line transformation of An. albimanus.
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  • 25
    ISSN: 1572-994X
    Keywords: adenovirus type 12 ; E1A ; transformation ; p300 binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The early region 1A (E1A) gene of highly oncogenic adenovirus type 12 (Ad12) was analyzed for transforming activity and protein binding using specific mutations. The Ad12 E1A proteins were found to bind p300 protein mainly within the CR1 region, although mutations that affect both p300 binding and transformation were identified in both the CR1 and the N-terminal region. The most critical mutation dlf89 located in the CR1 region was further dissected by point mutations and the results identified 68S as the most critical for transformation and 67E as the most critical for p300 binding. Specific mutations that retain p300 binding but impair transcriptional repression of a viral enhancer were also identified in both the N-terminal and CR1 regions.
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  • 26
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    Euphytica 96 (1997), S. 153-161 
    ISSN: 1573-5060
    Keywords: cassava ; transformation ; review ; somatic embryogenesis ; adventitious shoot formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A prerequisite for the development of a successful transformation system is the availability of efficient regeneration systems. Up to 1995 the only available regeneration system in cassava was an organized type of somatic embryogenesis. Transformation of these organized somatic embryogenic cultures with particle bombardment or Agrobacterium tumefaciens resulted in chimeric transformed embryos. However, the transformed sector was lost after repeated cycles of secondary somatic embryogenesis. After 1995 a less organized system of somatic embryogenesis was developed, so called friable embryogenic callus (FEC) and a system of adventitious shoot regeneration. The FEC regeneration system was combined successfully with particle bombardment. Selection of transgenic plants was based on either luciferase activity, or resistance to the aminoglycoside paromomycin or the herbicide phosphinothricin. Furthermore, protoplasts of FEC are able to regenerate into plants and can be transformed by electroporation. The adventitious shoot regeneration system was combined successfully with Agrobacterium tumefaciens. For this mature somatic embryos were cocultivated with Agrobacterium and cultured for adventitious shoot development. After selection based on the aminoglycoside geneticin or on hygromycin transgenic plants were formed.
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  • 27
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    Euphytica 96 (1997), S. 167-172 
    ISSN: 1573-5060
    Keywords: Coffea spp. ; coffee ; genetic ; in vitro ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The most important advances obtained on in vitro coffee regeneration systems and in coffee genetic transformation, drawing perspectives and scopes to further studies in these fields are presented and discussed.
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  • 28
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    Plant molecular biology 35 (1997), S. 205-218 
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; rice ; transformation ; monocotyledons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Agrobacterium tumefaciens has been routinely utilized in gene transfer to dicotyledonous plants, but monocotyledonous plants including important cereals were thought to be recalcitrant to this technology as they were outside the host range of crown gall. Various challenges to infect monocotyledons including rice with Agrobacterium had been made in many laboratories, but the results were not conclusive until recently. Efficient transformation protocols mediated by Agrobacterium were reported for rice in 1994 and 1996. A key point in the protocols was the fact that tissues consisting of actively dividing, embryonic cells, such as immature embryos and calli induced from scutella, were co-cultivated with Agrobacterium in the presence of acetosyringonc, which is a potent inducer of the virulence genes. It is now clear that Agrobacterium is capable of transferring DNA to monocotyledons if tissues containing ‘competent’ cells are infected. The studies of transformation of rice suggested that numerous factors including genotype of plants, types and ages of tissues inoculated, kind of vectors, strains of Agrobacterium, selection marker genes and selective agents, and various conditions of tissue culture, are of critical importance. Advantages of the Agrobacterium-mediated transformation in rice, like on dicotyledons, include the transfer of pieces of DNA with defined ends with minimal rearrangements, the transfer of relatively large segments of DNA, the integration of small numbers of copies of genes into plant chromosomes, and high quality and fertility of transgenic plants. Delivery of foreign DNA to rice plants via A. tumefaciens is a routine technique in a growing number of laboratories. This technique will allow the genetic improvement of diverse varieties of rice, as well as studies of many aspects of the molecular biology of rice.
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  • 29
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    Water, air & soil pollution 95 (1997), S. 35-43 
    ISSN: 1573-2932
    Keywords: humic acid ; mercury ; transport ; transformation ; soil-plant system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The influence of humic acid (HA) on the transport and transformation of mercury (Hg) in soil was studied. No available Hg could be detected (〈2.5 μg kg−1) in alluvial soil when the content of HA-carbon (HA-C) was higher than 0.2 g kg−1 although a large amount of Hg (8 μg kg−1) was applied to the soil. The available Hg decreased with the increase of HA in purple soil (r=0.735). There are significant correlations between HA concentration and organic Hg in the tested soils (r=0.974 for the purple soil and r=0.979 for the alluvial soil). The increase of HA results in decrease of Hg absorbed by plant from the soil. A loss of Hg from soil caused by microbes was observed.
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  • 30
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    Water, air & soil pollution 95 (1997), S. 35-43 
    ISSN: 1573-2932
    Keywords: humic acid ; mercury ; transport ; transformation ; soil-plant system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The influence of humic acid (HA) on the transport and transformation of mercury (Hg) in soil was studied. No available Hg could be detected (〈2.5 μg kg-1) in alluvial soil when the content of HA-carbon (HA-C) was higher than 0.2 g kg-1 although a large amount of Hg (8 μg kg-1) was applied to the soil. The available Hg decreased with the increase of HA in purple soil (r=0.735). There are significant correlations between HA concentration and organic Hg in the tested soils (r=0.974 for the purple soil and r=0.979 for the alluvial soil). The increase of HA results in decrease of Hg absorbed by plant from the soil. A loss of Hg from soil caused by microbes was observed.
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  • 31
    ISSN: 1573-2932
    Keywords: desorption ; ferrihydrite ; modelling ; phosphate ; TEM ; transformation ; wastewater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The presence of phosphate retards the transformation of ferrihydrite into crystalline products. Increasing phosphate from 0 to 1 mole % results in an order of magnitude decrease in the rate of transformation of ferrihydrite at pH 12. Levels of phosphate of ∼1 mol % suppress the formation of goethite (α-FeO(OH)) and result in the formation of a product consisting of η-Fe2O3. Higher levels of phosphate result in the ferrihydrite remaining amorphous, even after several hundred hours. Phosphate prevents formation of goethite by hindering the dissolution of ferrihydrite rather than by interfering with nucleation and growth of goethite in solution. The transformation rate of pure ferrihydrite is also strongly inhibited in the presence of dissolved phosphate. This is due to surface complexation. The transformation rate was measured at temperatures of 60 °C and 70 °C. The rate of transformation was found to be described by either (i) a solid-state reaction equation for powdered compacts or (ii) a zero-order reaction controlled by desorption. The transformation of the ferrihydrite matrix was accompanied by the loss of the phosphate trace component. X-ray diffraction indicates that no solid solution involving phosphate substitution into η-Fe2O3 is formed. Transmission electron microphotographs of the original precipitates containing phosphate confirm the presence of the phosphate and demonstrate its involvement in linking together extremely small particles of ferrihydrite.
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  • 32
    ISSN: 1573-2932
    Keywords: desorption ; ferrihydrite ; modelling ; phosphate ; TEM ; transformation ; wastewater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The presence of phosphate retards the transformation of ferrihydrite into crystalline products. Increasing phosphate from 0 to 1 mole % results in an order of magnitude decrease in the rate of transformation of ferrihydrite at pH 12. Levels of phosphate of ∼1 mol % suppress the formation of goethite (α-FeO(OH)) and result in the formation of a product consisting ofη-Fe2O3. Higher levels of phosphate result in the ferrihydrite remaining amorphous, even after several hundred hours. Phosphate prevents formation of goethite by hindering the dissolution of ferrihydrite rather than by interfering with nucleation and growth of goethite in solution. The transformation rate of pure ferrihydrite is also strongly inhibited in the presence of dissolved phosphate. This is due to surface complexation. The transformation rate was measured at temperatures of 60 °C and 70 °C. The rate of transformation was found to be described by either (i) a solid-state reaction equation for powdered compacts or (ii) a zero-order reaction controlled by desorption. The transformation of the ferrihydrite matrix was accompanied by the loss of the phosphate trace component. X-ray diffraction indicates that no solid solution involving phosphate substitution intoη-Fe2O3 is formed. Transmission electron microphotographs of the original precipitates containing phosphate confirm the presence of the phosphate and demonstrate its involvement in linking together extremely small particles of ferrihydrite.
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  • 33
    ISSN: 1572-9788
    Keywords: Agrobacterium tumefaciens ; bar gene ; grain legume ; Lupinus angustifolius ; phosphinothricin ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Transgenic plants of Lupinus angustifolius L. (cvs. Unicrop and Merrit) were routinely generated using Agrobacterium-mediated gene transfer to shoot apices. The bar gene for resistance to phosphinothricin (PPT, the active ingredient of the herbicide Basta) was used as the selectable marker. After co-cultivation, the shoot apex explants were transferred onto a PPT-free regeneration medium and their tops were thoroughly wetted with PPT solution (2 mg/ml). The multiple axillary shoots developing from the shoot apices were excised onto a medium containing 20 mg/l PPT. The surviving shoots were transferred every second week onto fresh medium containing 20 mg/l PPT. At each transfer, the number of surviving shoots decreased, until it stabilized. Indeed, some of these chimeric shoots surviving the PPT selection, eventually produced new green healthier axillary shoots which could be transferred to soil. This whole process took from 5 to 9 months after co-cultivation. Average transformation frequencies of 2.8% for cv. Unicrop and of 0.4% for the commercial cultivar Merrit were achieved. Molecular analysis of T0, T1, and T2 generations demonstrated stable integration of the foreign gene into the plant genome and expression of the integrated gene. Transformed plants of the T1 and T2 generations were resistant in glasshouse trials where the herbicide Basta (0.1 mg/ml) was sprayed onto whole plants. These results demonstrate that Agrobacterium-mediated gene transfer to preorganised meristematic tissue combined with axillary regeneration can form the basis of a routine transformation system for legume crop species which are difficult to regenerate from other explants.
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  • 34
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    World journal of microbiology and biotechnology 14 (1997), S. 271-276 
    ISSN: 1573-0972
    Keywords: Biodegradation ; 3-chlorobenzoic acid ; degradative plasmid ; Pseudomonas putida ; rec− Escherichia coli DH5 ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 35
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    World journal of microbiology and biotechnology 14 (1997), S. 7-10 
    ISSN: 1573-0972
    Keywords: Biodegradation ; morpholine ; plasmid ; Pseudomonas fluorescens ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A fast-growing Pseudomonas fluorescens CAS102, isolated by enrichment technique from polluted soil, effectively utilized morpholine as the energy source. The strain was able to grow in high concentrations of morpholine but accumulation of ammonia inhibited its growth and complete mineralization. The molar conversion ratio of morpholine to ammonia was 1:0.82. The organism harboured a single, multiple antibiotic- and heavy metal-resistance 140kb plasmid which was resistant to curing. Transformation studies showed that the morpholine degradative phenotype was expressed only in Pseudomonas putida and not in Escherichia coli. Growth studies on different degradative intermediates of morpholine suggested that plasmid-encoded genes were involved in the heterocyclic ring cleavage and the remaining reactions were mediated by chromosomal genes.
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  • 36
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    Biologia plantarum 40 (1997), S. 507-514 
    ISSN: 1573-8264
    Keywords: herbicide resistance ; PCR ; Phaseolus vulgaris ; regeneration ; Southern blotting ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We obtained transformed bean shoots by electroporation of intact bean cells with the plasmid pDPG165 containing bar gene conferring herbicide resistance to plants. Transformed shoots were selected from electroporated callus on herbicide containing media. Data of molecular analysis (PCR and Southern blotting) confirmed the insertion of bar gene in the genome of herbicide resistant shoots. Detailed procedures for obtaining regenerative bean callus, optimization of electroporation of intact cells and transgenic shoots are given.
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  • 37
    ISSN: 1573-9368
    Keywords: β-GUS ; embryogenesis reinduction ; kanamycin ; NPTII ; Piceaglauca ; selection ; somatic embryos ; somaticseedlings ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformed white spruce cultures containing immature Stage I somatic embryos, were developed after particle bombardment of somatic embryos with pBI 426, carrying an expression cassette with a gus::nptII fusion gene. These Stage I cultures did not show tolerance to kanamycin concentrations above 3 to 5 mg l−1, although assays for GUS and NPTII showed that functional enzymes were present in the transgenic tissue. Embryonic liquid suspension cultures were initiated from this transformed tissue. After treatment on agar-solidified maturation medium with 48 μm (±)-ABA high numbers of Stage III (cotyledonary) somatic embryos were produced. When subjected to an embryogenesis re-induction process with 2,4-D and BA, these Stage III embryos produced a new generation of Stage I embryogenic tissues which could tolerate 5--10 mg l−1 kanamycin. Stage III somatic embryos could alternatively be placed onto germination medium for the development of somatic seedlings. When germinated in the presence of 20 mg l−1 kanamycin, 77% of inoculants were resistant. The stability of integration of the gus::nptII fusion gene in the genome of white spruce Stage III somatic embryos and somatic seedlings was confirmed through Southern blot analysis
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  • 38
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    Transgenic research 6 (1997), S. 329-336 
    ISSN: 1573-9368
    Keywords: Agrobacterium ; SAAT ; sonication ; transformation ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plant transformation via Agrobacterium can be limited by both host specificity and the inability of Agrobacterium to reach the proper cells in the target tissue. Described here is a new and efficient Agrobacterium-based transformation technology that overcomes these barriers and enhances DNA transfer in such diverse plant groups as dicots, monocots, and gymnosperms. This new technology, called sonication-assisted Agrobacterium-mediated transformation (SAAT), involves subjecting the plant tissue to brief periods of ultrasound in the presence of Agrobacterium. Scanning electron and light microscopy reveal that SAAT treatment produces small and uniform fissures and channels throughout the tissue allowing the Agrobacterium easy access to internal plant tissues. Unlike other transformation methods, this system has the potential to transform meristematic tissue buried under several cell layers. SAAT increases transient transformation efficiency in several different plant tissues including leaf tissue, immature cotyledons, somatic and zygotic embryos, roots, stems, shoot apices, embryogenic suspension cells and whole seedlings. A 100- to 1400-fold increase in transient β- glucuronid ase expression has been demonstrated in various tissues of soybean, Ohio buckeye, cowpea, white spruce, wheat and maize. Stable transformation of both soybean and Ohio buckeye has been obtained using SAAT of embryogenic suspension culture tissues. For soybean, SAAT treatment was necessary to obtain stable transformation with this tissue
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  • 39
    ISSN: 1573-9368
    Keywords: GUS ; matrix attachment regions ; Populus ; transformation ; transgene expression ; Agrobacterium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We tested the value of a matrix attachment region (MAR) fragment derived from a tobacco gene for increasing the frequency of Agrobacterium-mediated transformation. A binary vector that carried a GUS reporter gene containing an intron and an nptII gene was modified to contain flanking MAR elements within the T-DNA borders. Vectors containing or lacking MARs were then used to transform tobacco, a readily transformabl e poplar clone (Populus tremula × P. alba), and a recalcitrant poplar clone (Populus trichocarpa × P. deltoides). MARs increased GUS gene expression approximately 10-fold in the two hybrid poplar clones and twofold in tobacco one month after cocultivation with Agrobacterium; MARs also increased the frequency of kanamycin-resistant poplar shoots recovered
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