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  • 1995-1999  (31)
  • 1980-1984  (4)
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  • 1
    Electronic Resource
    Electronic Resource
    rol-Transgenic Populus tremula: root development, root-borne bud regeneration and in vitro propagation efficiency (1998)
    Springer
    Trees 12 (1998), S. 464-471 
    Details
    ISSN: 0931-1890
    Keywords: Key words Agrobacterium rhizogenes ; Populus tremula ; rol Genes ; Root formation ; Transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  The potential use of the rol genes from Agrobacterium rhizogenes to improve the root system horticultural characteristics was evaluated in transgenic aspen (Populus tremula) plants, harboring the rol genes under their native promoters. Southern blot and RT-PCR analyses confirmed the presence and expression of A. rhizogenes rolC and rolB genes in four different phenotypically selected transgenic clones. Several of the observed phenotypic modifications were related to rol-gene expression and included, in particular, modified root systems. All in vitro-cultured rol-transgenic plants exhibited extensive root formation in a hormone-free medium, as well as a larger root surface area and mass, as compared to a uidA (β-glucuronidase-encoding)-transgenic aspen line and control (non-transformed) plants. Adventitious root formation in stem segments of rol-transgenic plants exhibited very rapid kinetics, resulting in a much shorter rooting time for rol-transgenic stem segments (e.g. 10 days for 80% rooting in rol-transgenic lines T-26 and T-27, as compared to more than 18 days for control non-transformed or uidA-transgenic aspen plants). rol-Transgenic plants maintained the capacity for 100% rooting throughout the year, versus 70–80% rooting in non-transformed plants during the winter. The four rol-transgenic lines exhibited differences in root development; in two of them enhanced root development was accompanied by increased shoot fresh weight. The root:shoot fresh weight ratio was always higher in rol-transgenic lines than in non-transformed plants. In the T-27 rol-transgenic line, the propagation coefficient of shoot-bud regeneration in liquid root culture was almost three times higher than in non-transformed plants. To the best of our knowledge this is the first report on quantitative phenotypic alterations in rol-transgenic woody plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00009729
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  • 2
    Electronic Resource
    Electronic Resource
    rol-Gene expression in transgenic aspen (Populus tremula) plants results in accelerated growth and improved stem production index (1999)
    Springer
    Trees 14 (1999), S. 49-54 
    Details
    ISSN: 0931-1890
    Keywords: Key words Transgenic poplar ; Agrobacterium rhizogenes ; Populus tremula ; rol genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  Stem and trunk growth, axillary bud break and branching habits are extremely important parameters of wood production in forest trees. The possibility of altering tree form by transformation with genes responsible for hormone biosynthesis and/or activity is most attractive. We examined four different phenotypically selected transgenic clones of a model tree –Populus tremula– expressing rol genes from Agrobacterium rhizogenes under their native promoters. Several of the observed phenotypic modifications were correlated with rol-gene expression, including breaking of stem apical dominance which resulted in the development and branching of up to four axillary buds per explant, as compared to a lack of axillary bud break in a uidA (β-glucuronidase-encoding)-transgenic aspen line and control (non-transformed) plants. rol-Transgenic plants also exhibited a higher cumulative stem length and enhanced growth rate, and hence a higher stem production index. During their first and second years in the greenhouse, rol-transgenic aspen plants exhibited enhanced growth and delayed winter dormancy relative to non-transformed plants. Although initially rol-transgenic plants had smaller, wrinkled leaves, these changes were not observed in the 2-year-old plants, which exhibited a phenotypically true-to-type leaf shape.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00009753
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  • 3
    Electronic Resource
    Electronic Resource
    Channels in the Gramicidin S-with-Urea Structure and their Possible Relation to Transmembrane Ion Transport (1997)
    Copenhagen : International Union of Crystallography (IUCr)
    Acta crystallographica 53 (1997), S. 151-159 
    Details
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: The structure of membrane-active antibiotic cyclodecapeptide gramicidin S in the crystals of its complex with urea, C60H92N12010.0.5[(NH2)2CO].7.94H20, has been investigated with three-dimensional X-ray data by the automatic sequential approximation method. The crystals are trigonal, space group P3121, a = 25.80(3), c= 21.49 (2) Å, Mr = 7968, calculated density = 1.088 mg m−3, Z = 1. Conventional R factor: R1 = 0.0943, wR2 = 0.2478 [I〉 2σ(I)]. The molecule possesses an antiparallel twisted β-structure, with turns involving the Phe-Pro peptides. The Orn side chains extend on one side of the sheet, while the non-polar Val and Leu side chains are located on the other face. One of the Orn residues (namely Orn2) is linked by an intermolecular hydrogen bond to the O atom of Phe4 residue, the other is free. The side chains of the Phe residues have trans orientation (χ1 ∼ 180°) and those of the Val, Orn, Leu residues, except those of Orn2, have the preferential gauche orientation with the χ1 angle close to 60. Two side chains show statistical disorder and conformation of the Pro residues is Cs—Cβ-exo. There is half a urea molecule and also 7.94 water molecules distributed on 13 positions for each antibiotic molecule. A partially occupied and poorly ordered alcohol molecule had been identified. The gramicidin S molecules are arranged around the 31 axis in the form of a left-handed double spiral forming suggestive channels. The outer hydrophobic surface of the spiral is made of uncharged side radicals while the inside surface consists of the main-chain atoms, mainly O and N, and of ornithine side chains with N atoms at the ends. By changing the Orn side-chain conformation, the inner diameter of the channels may change from 3.4 to 6.3 Å. Thus, ions and particles of rather large size may pass through the channel. The possibility of the creation of the gramicidin S channels in mitochondrial membranes has been noted by some biochemists. The channel complexes are close-packed in a hexagonal arrangement in the crystal. The CI− ions, present in abundance in the mother solution, are not found ordered in the crystals, which may indicate the absence of the charges in the terminal N atoms of the Orn residues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1107/S0907444995000916
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  • 4
    Electronic Resource
    Electronic Resource
    High frequency gene transfer by microprojectile bombardment of intact conidia from the entomopathogenic fungus Paecilomyces fumosoroseus (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 156 (1997), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Two different methods, (i) PEG and (ii) biolistic, were employed to transform protoplasts and conidia of Paecilomyces fumosoroseus using hygromycin resistance as selectable marker. Transformation frequencies varied from 1.9 to 2.5 transformants μg−1 of DNA by the PEG method, and from 33 to 153 transformants μg−1 of DNA by the biolistic procedure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1997.tb12711.x
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  • 5
    Electronic Resource
    Electronic Resource
    Genetic transformation of germinated conidia of the thermophilic fungus Humicola grisea var. thermoidea to hygromycin B resistance (1998)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 169 (1998), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Germinated conidia of the thermophilic fungus Humicola grisea var. thermoidea were transformed to hygromycin B resistance using the plasmid pAN7.1. Transformation was achieved using lithium acetate treatment or electroporation. The efficiency of transformation was up to 32 and 25 transformants per μg of plasmid DNA with the two methods, respectively. Transformants obtained by the lithium acetate method were more stable and showed a high copy number of the hph gene integrated into their genome. The other transformants, from the electroporation procedure, were stable, but unable to grow in the presence of high levels of hygromycin, and detection of the hph gene was only possible by polymerase chain reaction analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1998.tb13316.x
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  • 6
    Electronic Resource
    Electronic Resource
    High frequency gene conversion among benomyl resistant transformants in the entomopathogenic fungus Metarhizium anisopliae (1996)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 142 (1996), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Three different methods, (i) PEG, (ii) electroporation and (iii) biolistic, were employed to transform Metarhizium anisopliae using benomyl resistance as a selectable marker. Transformation frequencies and mitotic stability varied for each method, from 0.8 to 6.9 transformants μg−1 of DNA and 46%, respectively, by the PEG method; 1.3 to 1.8 transformants μg−1 of DNA and 67% by electroporation; and 32 to 201 transformants μg−1 of DNA and 90% by biolistic. We demonstrate by PCR that 60% of the transformants were generated by gene conversion.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1996.tb08418.x
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  • 7
    Electronic Resource
    Electronic Resource
    Transformation of Aspergillus nidulans by microprojectile bombardment on intact conidia (1995)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 125 (1995), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract This paper describes transformation of intact conidia of Aspergillus nidulans, auxotrophic for arginine, by using the biolistic process. The plasmid employed was pFB39, carrying the argB gene. The transformation frequency obtained was 81 transformants/ μg of DNA. Classical genetics and molecular analysis were conducted to analyse transformants and to determine in which chromosome integration took place.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1995.tb07371.x
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  • 8
    Electronic Resource
    Electronic Resource
    Transformation and regeneration of transgenic aspen plants via shoot formation from stem explants (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 99 (1997), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: An Agrobacterium-mediated transformation procedure for aspen (Populus tremula L.), involving the direct regeneration of shoot-buds from stem explants, is described. Disarmed Agrobacterium tumefaciens strain EHA101 harboring the binary plasmid pKIW1105 (which carries the uidA and nptII genes, coding for β-glucuronidase [GUS] and neomycin phosphotransferase II, respectively) was used for the transformation of stem explants. An incubation period of 48 to 72 h was found to be most effective in terms of transient GUS expression on the cut surface of the stem explants. Adventitious shoots regenerated after 2–3 weeks of culture in a woody plant medium (WPM) supplemented with TDZ (1-phenyl-3-[1,2,3-thiadiazol-5-yl]-urea, Thidiazuron) and carbenicillin. Three different kanamycin-based selection schemes were evaluated for optimization of transformation efficiency: (1) Kanamycin was added only to the rooting medium (5 to 6 weeks post-inoculation), or (2) to the regeneration medium 10–14 days after inoculation, or (3) after 2 days of co-cultivation. The third selection scheme was found to be optimal for adventitious shoots with regard to both the time required and the transformation efficiency, the latter being much higher than with the other schemes. Leaf samples from kanamycin-resistant shoots and plantlets were tested for GUS expression, and subjected to polymerase chain reaction (PCR) analysis of uidA and nptII genes. A Southern blot of the corresponding PCR-amplified fragments confirmed their authenticity and Southern blots of total plant DNA confirmed integration of the nptII gene into the plant genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1997.tb05356.x
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  • 9
    Electronic Resource
    Electronic Resource
    Agrobacterium rhizogenes-mediated DNA transfer inPinus halepensis Mill. (1996)
    Springer
    Plant cell reports 16 (1996), S. 26-31 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Agrobacterium rhizogenes strain LBA9402 was used to transformPinus halepensis embryos, seedlings and shoots. Mature embryos exhibited susceptibility to the agrobacterium as monitored by β-glucurortidase (GUS) expression, with more than 85% showing considerable transient GUS expression in the radicle. GUS expression was also observed in cotyledons, but at a lower rate of about 24% of the embryos (1–5 spots/embryo). Stable transformation was evidenced by the regeneration of GUS-expressing roots and calli from infectedP. halepensis seedlings. Inoculum injections into intact seedling hypocotyls induced callus and root formation at the wound sites in 64% of the seedlings. Dipping seedling cuttings in a bacterial suspension resulted in adventitious root formation in 7I% of the seedling cuttings, all of which expressed GUS activity. Adventitious shoots, that were induced on 2.5-year-old seedlings by pruning and spraying with 6-benzylaminopurine, were infected by injecting of bacterial suspension into their basal side. Two months later, adventitious roots and root primordia regenerated in 74% and 40% of 2- and 5-month-old shoots, respectively. Non-transformed shoots, either without or with auxin application, failed to form roots. Polymerase chain reaction and Southern blot analyses confirmed theuidA-transgenic nature of the root and callus, as well as the presence ofrolC androlB genes in roots from infectedP. halepensis seedlings.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01275443
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  • 10
    Electronic Resource
    Electronic Resource
    Agrobacterium rhizogenes-mediated DNY transfer in Pinus halepensis Mill. (1996)
    Springer
    Plant cell reports 16 (1996), S. 26-31 
    Details
    ISSN: 1432-203X
    Keywords: Abbreviations: BA, 6-benzylaminopurine; NOS, nopaline synthase; PCR, polymerase chain reaction; EtOH, ethanol; GUS, β-glucuronidase; NPTII, neomycin phosphotransferase II; CaMV, califlower mosaic virus; X-gluc, 5-bromo-4-chloro-3-indolyl β-D-glucuronic acid.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Agrobacterium rhizogenes strain LBA9402 was used to transform Pinus halepensis embryos, seedlings and shoots. Mature embryos exhibited susceptibility to the agrobacterium as monitored by β-glucuronidase (GUS) expression, with more than 85% showing considerable transient GUS expression in the radicle. GUS expression was also observed in cotyledons, but at a lower rate of about 24% of the embryos (1–5 spots/embryo). Stable transformation was evidenced by the regeneration of GUS-expressing roots and calli from infected P. halepensis seedlings. Inoculum injections into intact seedling hypocotyls induced callus and root formation at the wound sites in 64% of the seedlings. Dipping seedling cuttings in a bacterial suspension resulted in adventitious root formation in 71% of the seedling cuttings, all of which expressed GUS activity. Adventitious shoots, that were induced on 2.5-year-old seedlings by pruning and spraying with 6-benzylaminopurine, were infected by injecting of bacterial suspension into their basal side. Two months later, adventitious roots and root primordia regenerated in 74% and 49% of 2- and 5-month-old shoots, respectively. Non-transformed shoots, either without or with auxin application, failed to form roots. Polymerase chain reaction and Southern blot analyses confirmed the uidA-transgenic nature of the root and callus, as well as the presence of rolC and rolB genes in roots from infected P. halepensis seedlings.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050169
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