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  • Brassica napus
  • gene transfer
  • Springer  (32)
  • American Physical Society
  • Institute of Physics
  • MDPI Publishing
  • Periodicals Archive Online (PAO)
  • Wiley-Blackwell
  • 2000-2004  (24)
  • 1955-1959  (8)
  • 1940-1944
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  • Springer  (32)
  • American Physical Society
  • Institute of Physics
  • MDPI Publishing
  • Periodicals Archive Online (PAO)
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  • 1
    Electronic Resource
    Electronic Resource
    The spatio-temporal distribution of adult Ceutorhynchus assimilis in a crop of winter oilseed rape in relation to the distribution of their larvae and that of the parasitoid Trichomalus perfectus (2000)
    Springer
    Entomologia experimentalis et applicata 95 (2000), S. 161-171 
    Details
    ISSN: 1570-7458
    Keywords: Ceutorhynchus assimilis ; Trichomalus perfectus ; Brassica napus ; cabbage seed weevil ; parasitoid ; oilseed rape ; spatio-temporal distribution ; SADIE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The spatio-temporal distribution of Ceutorhynchus assimilis Payk. (Coleoptera: Curculionidae) adults caught in a rectangular grid of flight traps in a crop of winter oilseed rape (Brassica napus L.) was mapped and was analysed using Spatial Analysis by Distance IndicEs (SADIE). Their distribution was compared to that of their larvae and that of their parasitoid Trichomalus perfectus (Walker) (Hymenoptera: Pteromalidae) in pods. The distribution of immigrating C. assimilis adults was consistent with their arrival at the crop boundaries and movement within the crop towards its centre. Adult C. assimilis were aggregated at all times, invasion being on two fronts, leading to the formation of two major clusters within the crop. Large areas of the crop remained relatively unpopulated. During the emigration phase, numbers declined simultaneously in all parts of the crop. The distributions of adult and larval C. assimilis and of larval T. perfectus were spatially associated. The distribution of the parasitoid did not show a density dependent relationship with that of its host. We discuss the movements of insects which underlie their population distributions, the value of integrating spatial information into improved management strategies for C. assimilis and the potential for the spatial targeting of insecticides to reduce the amount applied and to conserve T. perfectus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1004081704932
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  • 2
    Electronic Resource
    Electronic Resource
    Rapid-freeze fixation and substitution improves tissue preservation of microspores and tapetum in Brassica napus (2000)
    Springer
    Sexual plant reproduction 12 (2000), S. 237-240 
    Details
    ISSN: 1432-2145
    Keywords: Key words Freeze-substitution ; Transmission electron microscopy ; Brassica napus ; Tapetum ; Microspore ; Immunogold
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The method of rapid freeze-fixation and substitution was used with Brassica napus floral bud material in order to improve the preservation of microspore and tapetal organelle structure. When observed using transmission electron microscopy, the appearance of the freeze-substituted material differs in a number of ways from the chemically-fixed material previously studied, in particular for the lipid-rich elaioplasts and tapetosomes in the tapetal cells. The tapetosomes have a very electron-dense, opaque appearance when visualized after rapid fixation. In addition, we were able to observe other cytoplasmic details such as pockets in the endoplasmic reticulum and cytoskeletal structures such as microfilaments. Extracellular material was also well-preserved; for example, the fibrous material in the baculae of the developing microspore exine was also visible. Finally, in the freeze-fixed sections specific structures such as elaioplasts could be labelled by antibodies, which indicates that this method preserved protein epitopes that were destroyed by chemical fixation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004970050007
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  • 3
    Electronic Resource
    Electronic Resource
    A Brassica napus skp1-like gene promoter drives GUS expression in Arabidopsis thaliana male and female gametophytes (2000)
    Springer
    Sexual plant reproduction 13 (2000), S. 29-35 
    Details
    ISSN: 1432-2145
    Keywords: Key words Gene-specific expression ; skp1-like gene ; GUS staining ; Gametophytic expression ; Brassica napus ; Arabidopsis thaliana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We isolated a gene, BnSKP1γ1, expressed in rapeseed (Brassica napus) microspores, which encodes a protein closely related to the Saccharomyces cerevisiae Skp1p protein previously shown to play a role in cell cycle regulation. Twelve SKP1-related genes have already been identified in the Arabidopsis thaliana genome. Using a PCR-based strategy, we isolated three other genes. To date, most data available concerning the function of the SKP1-related genes in plants are indirect. Studies on transgenic A. thaliana plants showthat a 1100-bp BnSKP1γ1 promoter fragment can direct GUS expression in female gametophytes soon after the first haploid mitosis and in male gametophytes from the tetrade stage. No GUS expression can be detected in sporophytic tissues. RT-PCR experiments suggest that this gene is expressed in a similar way in rapeseed. This is the first reported case of a gene exhibiting such an expression pattern in angiosperms.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00009839
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  • 4
    Electronic Resource
    Electronic Resource
    Effect of exogenous calcium on Agrobacterium tumefaciens-mediated gene transfer in Hevea brasiliensis (rubber tree) friable calli (2000)
    Springer
    Plant cell reports 19 (2000), S. 851-855 
    Details
    ISSN: 1432-203X
    Keywords: Key words Rubber tree ; Agrobacterium tumefaciens ; calcium ; gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The influence of CaCl2 was investigated on Agrobacterium tumefaciens-mediated gene transfer in Hevea brasiliensis friable calli which are usually proliferated on maintenance medium (MM) containing 9 mM CaCl2.Five A. tumefaciens strains (C58pMP90, C58pGV2260, AGL1, LBA4404 and EHA 105) and two binary vectors (pGIN and pCAMBIA2301) were tested and the strain EHA105pC2301 was selected to conduct further experiments. The calli were precultured on MM containing a range of CaCl2 concentrations, then inoculated with Agrobacterium suspension. Transfer of friable calli from MM containing 9 mM CaCl2 to calcium-free medium significantly enhanced the transient β-glucuronidase activity. Interestingly, the use of calcium-free Agrobacterium resuspension medium to inoculate friable calli again dramatically increased the transformation efficiency. Induction of Agrobacterium's virulence with acetosyringone remained an important factor to stimulate transformation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990000208
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  • 5
    Electronic Resource
    Electronic Resource
    Genetic analysis of shoot regeneration from cotyledonary explants in Brassica napus (2000)
    Springer
    Theoretical and applied genetics 100 (2000), S. 895-898 
    Details
    ISSN: 1432-2242
    Keywords: Key words Diallel analysis ; Regeneration ability ; Cotyledonary culture ; Brassica napus ; Heritablity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Genetic analysis of shoot regeneration from cotyledonary explants of Brassica napus was carried out by 7×7 diallel crosses using cultivars showing a different ability for regeneration. Both additive and dominant effects were significant, with the additive effect being more important than the dominant one. Dominant genes had a positive effect on shoot regeneration. Non-allelic interaction and average maternal effects were not detected, while specific the maternal one was significant. In the 5×5 sub-diallel table, the maternal effect became nonsignificant. The mean degree of dominance was 0.759. Broad- and narrow-sense heritabilities were 0.973 and 0.819, respectively, indicating that shoot regenera- tion ability can be easily transferred into economically important cultivars showing a low or an unresponsive ability.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051367
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  • 6
    Electronic Resource
    Electronic Resource
    Genetic analysis of fertility restoration and accumulation of ORF125 mitochondrial protein in the kosena radish (Raphanus sativus cv. Kosena) and a Brassica napus restorer line (2000)
    Springer
    Theoretical and applied genetics 100 (2000), S. 949-955 
    Details
    ISSN: 1432-2242
    Keywords: Key words. Fertility Restoration (Rf) ; Cytoplasmic male sterility (CMS) ; Raphanus sativus L. ; Brassica napus ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The genetics of fertility restoration (Rf) of kosena radish CMS has been characterized. The kosena CMS-Rf system is genetically the same as that of the ogura CMS-Rf system. Two dominant genes that act complementary to the restoration of fertility control fertility restoration in kosena CMS. One allele (Rf1) is associated with accumulation of the CMS-associated protein, ORF125. The interaction of Rf1 and another allele (Rf2) was essential for the restoration of fertility in radish, whereas Rf1 alone was sufficient for the complete restoration of fertility in the B. napus kosena CMS cybrid.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051375
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  • 7
    Electronic Resource
    Electronic Resource
    Spacer Length Dependence on the Efficiency of Dimeric Anionic Peptides in Gene Transfer by Glycosylated Polylysine/Plasmid Complexes (2000)
    Springer
    Bioscience reports 20 (2000), S. 383-398 
    Details
    ISSN: 1573-4935
    Keywords: fusogenic peptides ; HA2 influenza virus hemagglutinin ; polylysine ; transfection ; gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Amphiphilic anionic peptides have been used to enhance the efficiency of transfection by helping plasmids to escape from endosomes to the cytosol. It has been shown that efficiency of an eicosamers containing five glutamyl residues (E5), can be considerably enhanced either by transforming it into a dimer or by adding a tripeptide WYG in a C-terminal position (E5WYG). The dimerization of the peptide E5WYG leads to a more efficient tool when the dimerization device includes the tripeptide WYG unit and a longer spacer arm made of Gly-βAla-βAla residues, but to a 10-fold less efficient tool when the dimerization device includes a shorter spacer, a glycyl residue. Both dimers are taken up by the cells to a similar extent. Both dimers seem to be surrounded similarly as far as the environmental pH is concerned. In contrast, we found a correlation between the propensity of the peptides to adopt a helical structure at neutral pH and the gene transfer efficiency.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1010382001654
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  • 8
    Electronic Resource
    Electronic Resource
    Efficient Gene Delivery Using Anionic Liposome-Complexed Polyplexes (LPDII) (2000)
    Springer
    Bioscience reports 20 (2000), S. 419-432 
    Details
    ISSN: 1573-4935
    Keywords: liposomes ; gene therapy ; gene transfer ; nonviral vectors ; polylysine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Synthetic gene transfer vectors based on polyplexes complexed to anionic liposomes (LPDII vectors) were characterized for their transfection efficiency in cultured mammalian cells. The effects of polycation to DNA ratio, lipid to DNA ratio, choice of polycation and lipid composition were systematically evaluated in human oral carcinoma KB cells, using a luciferase reporter gene. For LPDII formulations containing poly-L-lysine and dioeoylphosphatidylethanolamine/cholesteryl hemisuccinate (DOPE/CHEMS) anionic liposomes, at a constant lipid to DNA ratio, an increase in the polycation/DNA (N/P) ratio resulted in an increase in transfection activity. Meanwhile, the optimal lipid to DNA ratio for efficient gene delivery was influenced by the N/P ratio used, and was increased at higher N/P ratios. For the DNA condensing agent, poly-L-lysine could be replaced by polyethylenimine (PEI) as the DNA condensing agent in the formulations. For the lipidic components, CHEMS could be replaced by other anioniclipids including oleic acid, dicetylphosphate and phosphatidylserine, but DOPE, a fusogenic helper lipid, could not be replaced by dioleolyphosphatidylcholine. LPDII formulation showed significantly less cytotoxicity compared to the commonly used cationic lipsomes or PEI mediated transfection and several cell lines were transfected with high efficiency. LPDII vectors avoid the use of toxic cationic lipids and may have potential application in gene therapy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1010338219401
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  • 9
    Electronic Resource
    Electronic Resource
    Direct somatic embryogenesis, plant regeneration and in vitro flowering in rapid-cycling Brassica napus (2000)
    Springer
    Plant cell reports 19 (2000), S. 1177-1183 
    Details
    ISSN: 1432-203X
    Keywords: Keywords Rapid-cycling ; Brassica napus ; Somatic embryogenesis ; Secondary embryogenesis ; Regeneration ; In vitro Howering ; AbbreviationsABA: Abscisic acid ; BAP: 6-Benzyl-aminopurine ; DAP: Days after pollination ; 2-iP: 6-(γ, γ-dimethlyallyl-amino)purine ; Kinetin: 6-Furfurylaminopurine ; MS: Murashige and Skoog ; SE0: Somatic embryo from seed ; SE1: First-generation secondary embryo ; SE2: Second-generation secondary embryo ; Zeatin: 6-(4-hydroxy-3-methylbut-trans-2-enylamino)purine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A simple method to induce somatic embryogenesis from seeds of rapid-cycling Brassica napus is described. Seedlings cultured on Murashige and Skoog (MS) basal medium produced somatic embryos directly on hypocotyls and cotyledons after 2 to 3 subcultures onto the same medium. A low pH of the medium (3.5–5) was more conducive to somatic embryogenesis than a higher pH (6 and 7). Embryogenic potential of the seeds was inversely correlated to seed age: about 41–68% of immature seeds between the ages of 14 and 28 days after pollination (DAP) formed somatic embryos compared to 0–11% of the seeds obtained 29–37 DAP. About 54% of the somatic embryos produced secondary embryos after subculturing onto the same medium. The embryogenic potential of the cultures has been maintained on MS basal medium for 2 years (12 generations) without diminution. Up to 75% of the secondary embryos developed into plantlets on MS medium enriched with 10–6  M zeatin, and 40% of these produced flowers when transferred to an optimised flower-induction medium. Viable seeds were produced in self-pollinated in vitro flowers.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990000268
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  • 10
    Electronic Resource
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    Insulin-like Growth Factor-I cDNA Gene Transfer in vitro and in vivo (2000)
    Springer
    Biochemical genetics 38 (2000), S. 41-55 
    Details
    ISSN: 1573-4927
    Keywords: gene transfer ; liposome ; IGF-I ; wound healing ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Our hypothesis is that gene transfer of an IGF-I CMV-cDNA with cholesterol containing cationic liposomes is an efficient tool for transient transfection of growth factors in vitro and in vivo. In vitro, we transiently cotransfected IGF-I cDNA with a CMV construct and a Lac Z CHKβCHK-galactosidase cDNA/CMV construct using cholesterol containing cationic liposomes and measured CHKβCHK-galactosidase and IGF-I mRNA and protein. In vivo, we subcutaneously injected 3-month-old male Sprague–Dawley rats with IGF-I cDNA and CHKβCHK-galactosidase cDNA into rat skin. After IGF-I and CHKβCHK-galactosidase were cotransfected into PC12 cells, Northern blot analysis showed that the peak time of IGF-I expression was 2 days for mRNA and 5 days for protein. In vivo, a cDNA/liposome ratio of 1:2 was most effective. IGF-I protein expression in IGF-I-transfected skin resulted in significant transfection from day 5 to day 7. In situ determination of CHKβCHK-galactosidase activity confirmed that transfections resulted in a restricted expression area.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1001880220039
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  • 11
    Electronic Resource
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    A Cationic Lipid Emulsion/DNA Complex as a Physically Stable and Serum-Resistant Gene Delivery System (2000)
    Springer
    Pharmaceutical research 17 (2000), S. 314-320 
    Details
    ISSN: 1573-904X
    Keywords: gene transfer ; lipid emulsions ; poly(ethylene glycol) ; transfection ; cationic lipids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To develop a non-viral gene delivery system in the form ofan oil-in-water (o/w) lipid emulsionMethod. Cationic lipid emulsions were formulated with soybean oil,1,2-dioleoyl-sn-glycero-3-trimethylammonium-propane (DOTAP) as acationic emulsifier and other co-emulsifiers. The physicalcharacteristics of the lipid emulsion and the emulsion/DNA complex weredetermined. The in vitro transfection efficiency of the emulsion/DNAcomplex was determined in the presence of up to 90% serum. Results. The average droplet size and zeta potential of emulsions wereca. 180 nm and ca. +50 mV, respectively. Among the emulsions, astable formulation was selected to form a complex with a plasmidDNA encoding chloramphenicol acetyltransferase. By increasing theratio of emulsion to DNA, zeta-potential of the emulsion/DNA complexincreased monotonously from negative to positive without any changesin the complex size. The complex was stable against DNase I digestionand an anionic poly-l-aspartic acid (PLAA). The complex deliveredDNA into the cells successfully, and the transfection efficiency wasnot affected by complex formation time from 20 min to 2 h. Moreimportantly, the cationic lipid emulsion facilitated the transfer of DNAin the presence of up to 90% serum. Conclusions. The cationic lipid emulsion/DNA complex has physicalstability and serum resistant properties for gene transfer.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007553106681
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  • 12
    Electronic Resource
    Electronic Resource
    Rearrangement of the actin filament and microtubule cytoskeleton during induction of microspore embryogenesis inBrassica napus L. cv. Topas (2000)
    Springer
    Protoplasma 213 (2000), S. 194-202 
    Details
    ISSN: 1615-6102
    Keywords: Actin microfilament ; Brassica napus ; Cytochalasin D ; Microspore embryogenesis ; Microtubule ; Preprophase band
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Changes in the actin filament and microtubule cytoskeleton were examined during heat- and cytochalasin D-induced embryogenesis in microspores ofBrassica napus cv. Topas by rhodamine phalloidin and immunofluorescence labelling respectively. The nucleus was displaced from its peripheral to a more central position in the cell, and perinuclear actin microfilaments and microtubules extended onto the cytoplasm. Heat treatment induced the formation of a preprophase band of microtubules in microspores; preprophase bands are not associated with the first pollen mitosis. Actin filament association with the preprophase band was not observed. The orientation and position of the mitotic spindle were altered, and it was surrounded with randomly oriented microfilaments. The phragmoplast contained microfilaments and microtubules, as in pollen mitosis I, but it assumed a more central position. Cytoskeletal reorganisation also occurred in microspores subjected to a short cytochalasin D treatment, in the absence of a heat treatment. Cytochalasin D treatment of microspores resulted in dislocated mitotic spindles, disrupted phragmoplasts, and symmetric divisions and led to embryogenesis, confirming that a normal actin cytoskeleton has a role in preventing the induction of embryogenesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01282157
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  • 13
    Electronic Resource
    Electronic Resource
    Application of in vitro organ culture in wide-cross breeding of rapeseed (2000)
    Springer
    Euphytica 114 (2000), S. 217-221 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; flower receptacle segments ; in vitro ; culture ; pollinated ovaries ; Raphanus sativus var. raphanistroides/kwd〉
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Oil radish (Raphanus sativus var. raphanistroides Makino) is resistant to drought and low temperature. In order to breed more resistant cultivars of rapeseed, the wide cross between rapeseed (Brassica napus L.) and oil radish was made. Rapeseed was not compatible with oil radish, and the frequency of hybrid plants (F1) was very low. Moreover, the hybrid plants were sterile. In order to recover the intergeneric hybrids (F1), the in vitro organ culture technique was applied in our experiments. The frequency of hybrid plants (F1) was increased up to 25.55% by means of in vitro culture of pollinated ovaries. Some fertile amphidiploid hybrid plants were obtained by means of colchicine treatment of small buds obtained from cultured flower receptacle segments of hybrid plants (F1). It is suggested that the technique of in vitro culture of pollinated ovaries and flower receptacle segments is useful in the wide-cross breeding of rapeseed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003911507540
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  • 14
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    In vitro selection for oleic and linoleic acid content in segregating populations of microspore derived embryos of Brassica napus (2000)
    Springer
    Euphytica 112 (2000), S. 195-201 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; doubled haploids ; high oleic acid rapeseed (HOAR) ; in vitro selection ; microsporeculture ; mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Microspore derived embryos (MDEs) in Brassica napuscontain large amounts of storage lipids which show a genotype specific fatty acid composition (FAC). One cotyledon of regenerating emblyos can be dissected at an early stage during the in vitro culture and used for fatty acid analysis. Thus, in breeding programmes to modify oil quality, only MDEs having the desired FAC need to be regenerated to plantlets and transferred to the greenhouse. In the present study the applicability of this method for the selection of a high oleic acid content and a low linoleic acid content in the seed oil has been tested by crossing a Brassica napus mutant line having a high oleic acid (C18:1) content in the seed oil (75%) with a wild type doubled haploid line with 62% C18:1 in the seed oil. Microspore culture was applied to the F1 plants. In total 59 MDEs were obtained, from which 31 were cultured with and 28 without 15μM abscisic acid for 3 weeksin vitro. One cotyledon was dissected under aspetic conditions and used for fatty acid analysis. The remaining part of the embryos were further regenerated to plantlets and transferred to the greenhouse to obtain seeds after self pollination. Seeds harvested from the doubled haploid lines in the greenhouse were used for fatty acid analysis and also for growing in the field. The abscisic acid treatment of the MDEs generally improved the correlations for linoleic and oleic acid between the MDEs and the seeds harvested in the greenhouse and the field. The correlations ranged from 0.68** to 0.81**.This indicates that selection for high oleic acid can be started already during an early stage of the in vitro culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003884823052
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  • 15
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    A comparison of methods for hybrid seed production using self-incompatibility in swedes (Brassica napus ssp. napobrassica) (2000)
    Springer
    Euphytica 113 (2000), S. 207-210 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; hybrid seed production ; self-incompatibility ; swedes, rutabaga
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Procedures for producing seed of hybrid swedes using self-incompatibility were examined. Single-cross, double-cross and modified double-cross hybrids were compared in isolation plots using natural pollinators and in polythene tunnels using blow-flies. With good coincidence of flowering and the same flower colour, nearly 100% hybrid seed was produced by natural pollinators with the single-crosses, the double-cross and one of the two modified double-cross hybrids; the other modified double-cross hybrid produced 87%hybrid seed. With poor coincidence of flowering and different flower colours the proportion of hybrids dropped to 61%. Using different flower colours and blow-flies as pollinators in polythene tunnels, higher levels of outcrossing were produced than in isolation plots with natural pollinators; the opposite result was obtained when the same flower colour was used.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003941606357
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  • 16
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    Rapid and efficient screening of phosphinothricin tolerant oilseed rape (Brassica napus) with a novel germination test (2000)
    Springer
    Euphytica 113 (2000), S. 119-124 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; germination test ; herbicidetolerance screening ; monitoring ; PAT-ELISA ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A novel screening test is described for the discrimination of transgenic phosphinothricin tolerant oilseed rape from non transgenic rape seedlings. The method is based on the germination of rape seeds on filter paper soaked with a 0.005% phosphinothricin solution. Under these conditions inhibition of seedling development by the herbicide can be observed after 10 days. The germination test gains an advantage over the routinely used herbicide spraying, because it is rapid, needs little space and allows efficient screening of huge numbers of seeds. The assay has been successfully applied to the screening of different transgenic and non transgenic rapeseed varieties/lines and has been compared to other methods such asBasta® spray test, drop test, ELISA-technique and PCR-amplification of the pat gene. This test allows on one hand large screening programmes to monitor the foreign gene in the environment and on the other quality control of seedlots before market introduction of herbicide tolerant oilseed rape.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003940106800
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  • 17
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    Chlorophyll breakdown in oilseed rape (2000)
    Springer
    Photosynthesis research 64 (2000), S. 137-146 
    Details
    ISSN: 1573-5079
    Keywords: Brassica napus ; chlorophyll ; chlorophyll catabolite ; degradation ; porphyrin ; senescence ; tetrapyrrole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chlorophyll catabolism accompanying leaf senescence is one of the most spectacular natural phenomena. Despite this fact, the metabolism of chlorophyll has been largely neglegted until recently. Oilseed rape has been used extensively as a model plant for the recent elucidating of structures of chlorophyll catabolites and for investigation of the enzymic reactions of the chlorophyll breakdown pathway. The key reaction which causes loss of green color is catalyzed in a two-step reaction by pheophorbide a oxygenase and red chlorophyll catabolite reductase. In this Minireview, we summarize the actual knowledge about catabolites and enzymes of chlorophyll catabolism in oilseed rape and discuss the significance of this pathway in respect to chlorophyll degradation during Brassica napus seed development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006456310193
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  • 18
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    Environmental and Developmental Regulation of Trypsin Inhibitor Activity in Brassica napus (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 1411-1422 
    Details
    ISSN: 1573-1561
    Keywords: Brassica napus ; defense-related proteins ; environmental effects ; induced defenses ; proteinase inhibitors ; regulation ; resistance ; trypsin inhibitors ; variation ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We examined several environmental and developmental influences on trypsin inhibitor (TI) activity in leaves of young Brassica napus seedlings in a series of greenhouse experiments. In seedlings of B. napus cv. Westar, TI activity is constitutively present and exhibits a rise then fall through time in the first true leaves of young plants. TI activity is induced by wounding in the first true leaves, but the degree of induction is relatively insensitive to the degree of wounding over a gradient of 5–15% of leaf area damage. TI activity is enhanced in first true leaves of plants in which the cotyledons have been wounded relative to plants in which the cotyledons have not been wounded. TI activity is also enhanced in the second true leaves on plants in which the first true leaves have been wounded. The degree of systemic induction in second true leaves declines additively with plant age, but local induction in the first true leaves is not affected by age. In B. napus cv. Gido, TI activity is constitutively present but is not locally wound-inducible in first true leaves of young plants exposed to the same wounding gradient as cv. Westar. In unwounded plants at the six-leaf stage, TI activity is higher in second true leaves than in fifth true leaves, indicating that TI activity is developmentally regulated in this cultivar.
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    URL: http://dx.doi.org/10.1023/A:1005540107957
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    Breeding to Increase the Concentration of 2-phenylethyl Glucosinolate in the Roots of Brassica napus (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 1811-1820 
    Details
    ISSN: 1573-1561
    Keywords: Brassica napus ; Pratylenchus neglectus ; nematode ; 2-phenylethyl glucosinolate ; isothiocyanate ; pest resistance ; disease break ; variability ; genetic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Root concentrations of 2-phenylethyl glucosinolate in canola, Brassica napus, influence the susceptibility of the crop to the root lesion nematode (Pratylenchus neglectus), as well as the nematicidal effect of root tissues as they degrade in the soil. Plants containing high 2-phenylethyl glucosinolate should therefore reduce soil populations of P. neglectus. A selection program was developed to increase the proportion of total glucosinolates contributed by 2-phenylethyl glucosinolate in the roots of B. napus cv. Dunkeld. Variation within this accession was observed to be stable across the S1 and S2 generations. The segregation observed for 2-phenylethyl glucosinolate percentage suggested that the trait was encoded at a single locus, with the "high" phenotype being dominant. Plants with the high 2-phenylethyl glucosinolate phenotype (〉45% of total glucosinolates) were shown to be significantly more resistant to P. neglectus than otherwise identical "low" phenotypes (〈45% of total).
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    URL: http://dx.doi.org/10.1023/A:1005588405774
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  • 20
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    Occurrence and diversity of plasmids in populations of streptomycetes in soil (2000)
    Springer
    Antonie van Leeuwenhoek 78 (2000), S. 323-329 
    Details
    ISSN: 1572-9699
    Keywords: gene transfer ; plasmids ; Streptomyces
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Studies were made of naturally occurring plasmids hosted in Streptomyces strains isolated from two different terrestrial ecosystems: an agricultural field and a protected forest area. Six out of the 147 screened isolates contained plasmids. The strains containing these plasmids were all isolated from the agricultural soil. Plasmids were not found among the strains isolated from the forest area. Cross hybridization of the six newly isolated plasmids revealed very high similarities between four of them. However, no similarities were found between the six newly isolated plasmids and well studied streptomycete plasmids such as pIJ101 and SCP2*. The host strains of the four similar plasmids belonged to three different species S. anulatus, S. rochei and S. diastaticus. This implies a possible conjugative transfer of these plasmids within the streptomycete population in the agricultural area. The reason for the absence of streptomycete plasmids from the populations derived from the forest area is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1010229215567
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  • 21
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    Effects of Agrobacterium tumefaciens-mediated transformation and tissue culture on storage lipids in Brassica napus (2000)
    Springer
    Euphytica 115 (2000), S. 181-190 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; fatty acid composition ; tissueculture-induced variation ; transformation-induced variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The variation obtained in storage fatty acids induced by the procedures of tissue culture and transformation with Agrobacterium tumefaciens was investigated and compared in rapeseed, Brassica napus, cv. Hanna. An increased variation in the fatty acid profiles was noted after tissue culture and transformation compared with plants derived directly from seeds. In the second generation of rapeseed transformants, T2, the content of oleic acid ranged from 39–72%, 12–31% for linoleic acid and 7–16% for linolenic acid. This could be compared with the oleic acid content in the T2 generation of tissue culture-derived plants which ranged between 47–76% and in seed-derived material where oleic acid ranged between 55–69%.In the T3 generation the ranges in transgenic seeds were decreased but still larger than in the seed derived plants. The range in transgenic plants was 49–64% for oleic acid, 20–28% for linoleic acid and 9–18% for linolenic acid. The most extreme individuals, both highest and lowest in the common fatty acids, were found in the group of transformed plants independent of generation. The total lipid content was also affected by the two treatments and seeds with the lowest and highest lipid content were both found among the transformed plants. In conclusion, care should be taken to use proper controls when performing transformation experiments in order to distinguish variation in the fatty acid profiles induced by the transformation procedure and tissue culture treatments from the changes due to transgenic expression.
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    URL: http://dx.doi.org/10.1023/A:1004026702787
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    Liposomes Containing Interferon-Gamma as Adjuvant in Tumor Cell Vaccines (2000)
    Springer
    Pharmaceutical research 17 (2000), S. 42-48 
    Details
    ISSN: 1573-904X
    Keywords: liposomes ; cancer vaccines ; cytokines ; immunotherapy ; interferon gamma ; gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. Liposomal systems may be useful as a cytokine supplementin tumor cell vaccines by providing a cytokine reservoir at the antigenpresentation site. Here, we examined the effect of liposomeincorporation of mIFNγ on its potency as adjuvant in an established tumor cellvaccination protocol in the murine B16 melanoma model. Adjuvanticityof the mIFNγ-liposomes was compared to that achieved bymIFNγ-gene transfection of the B16 tumor cells. Furthermore, we studiedwhether liposomal incorporation of mIFNγ indeed increases theresidence time of the cytokine at the vaccination site. Methods. C57B1/6 mice were immunized with i) irradiated IFNγ-genetransfected B16 melanoma cells or ii) irradiated wild type B16 cellssupplemented with (liposomal) mIFNγ, followed by a challenge withviable B16 cells. The residence time of the (liposomal) cytokine at thesubcutaneous (s.c.) vaccination site was monitored using radiolabeledmIFNγ and liposomes. Results. Immunization with irradiated tumor cells admixed withliposomal mIFNγ generated comparable protection against B16 challenge asimmunization with mIFNγ-gene modified tumor cells. Irradiated tumorcells admixed with soluble mIFNγ did not generate any protectiveresponses. Radiolabeling studies indicated that free mIFNγ rapidlycleared from the s.c. injection site. Association of [125I]-mIFNγwith liposomes increased the local residence time substantially: liposomalassociation of mIFNγ resulted in a prolonged local residence time ofthe cytokine as reflected by a 4-fold increase of the area under thecurve. The amount of released cytokine in the optimal dose rangecorresponds to the amount released by the gene-transfected cells. Moderate but significant CTL-activity against B16 cells was found for miceimmunized with irradiated cells supplemented with mIFNγ-liposomescompared to untreated control animals. Conclusions. Prolonged presence of mIFNγ at the site of antigenpresentation is crucial for the generation of systemic immune responsesin the B16 melanoma model. These studies show that liposomalencapsulation of cytokines is an attractive strategy for paracrine cytokinedelivery in tumor vaccine development.
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    URL: http://dx.doi.org/10.1023/A:1007514424253
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    Rhizopus and Fusarium are Selected as Dominant Fungal Genera in Rhizospheres of Brassicaceae (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 2387-2399 
    Details
    ISSN: 1573-1561
    Keywords: Rhizopus spp. ; Fusarium spp. ; rhizosphere microorganisms ; Brassicaceae ; Rorippa sylvestris ; Brassica napus ; Brassica juncea ; Lepidium sativum ; myrosinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We isolated several strains of Rhizopus and Fusarium spp. as dominant fungi in the rhizospheres of Brassicaceae plants. The Fusarium isolates showed a higher tolerance of the antifungal constituents of "mustard oil," which originates from the glucosinolates that are characteristic secondary metabolites of the Brassicaceae, than other Fusarium isolates from non-Brassicaceae plants. In contrast, the Rhizopus isolates showed a high tolerance regardless of their source. Myrosinase activity was found in Bn-R-1-1 (Rhizopus sp.) isolated from the rhizoplane of Brassica napus and Ls-F-in-4-1 (Fusarium sp.) isolated from a surface-disinfected root of Lepidium sativum (Brassicaceae). Ls-F-in-4-1 was the Fusarium most tolerant of the Brassicaceae antifungal constituents. These results suggest that fungi in the rhizospheres of Brassicaceae plants may be selected because of secondary metabolites exuded from the roots of host plants.
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    URL: http://dx.doi.org/10.1023/A:1005583012561
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    Expression of murine adenosine deaminase (ADA) in transgenic maize (2000)
    Springer
    Transgenic research 9 (2000), S. 1-9 
    Details
    ISSN: 1573-9368
    Keywords: gene transfer ; selectable marker ; adenosine analogues ; transgenic maize ; adenosine deaminase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A murine adenosine deaminase (ADA) gene, driven by the maize ubi-1 promoter and intron region, was transformed into embryogenic maize callus, along with a bar and gusA gene-containing plasmid, using microparticle bombardment. Selection in the presence of either the herbicide Basta® or the adenosine analogue 2′-deoxyadenosine resulted in transgenic cultures that expressed GUS and accumulated a 41kD protein that immunoprecipated with an ADA-specific polyclonal antibody. ADA enzyme activity was observed in extracts from transgenic callus as well as regenerated plants and progeny. Cultures expressing ADA grew in the presence of 200mg/l 2′-deoxyadenosine, a concentration which completely inhibited the growth of non-transgenic cultures. ADA activity appeared to segregate in progeny of regenerated plants as a single, dominant Mendelian trait. These results suggest that ADA, in combination with adenosine analogue selection, represents a potentially viable selectable marker system for transgenic maize production.
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    URL: http://dx.doi.org/10.1023/A:1008972101370
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  • 25
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    Shoot apical meristems as a target for gene transfer by microballistics (1955)
    Springer
    Euphytica 85 (1955), S. 45-51 
    Details
    ISSN: 1573-5060
    Keywords: meristem ; shoot apex ; ballistic microtargeting ; gene transfer ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The classical approach of gene transfer to a given plant species delivers the foreign gene to transformable cells and then puts the effort into generating plants. This approach is very difficult in many important crop plants, including cereals, and the results of regeneration are very genotype-dependent. In contrast, we use regenerable cells and try to transform them. Shoot apical meristems provide a tissue which regenerates in situ a fertile plant for most given genotypes or species. Transformation of meristem cells may lead to transgenic sectors in chimeras. These sectors may contribute to the gametes and, thus, to transgenic offspring, which then should be homohistonts and not sectorial chimeras like their parents. Our model plant for these studies is wheat. Microtargeting is a ballistic approach which is particularly suitable for the controlled delivery of microprojectiles to meristem cells in situ (Sautter et al., 1991). We summarize in this paper our experience with ballistic microtargeting of transgenes to wheat shoot apical meristem cells in situ.
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    URL: http://dx.doi.org/10.1007/BF00023929
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  • 26
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    A study of different (CaMV 35S and mas) promoter activities and risk assessment of field use in transgenic rapeseed plants (1955)
    Springer
    Euphytica 85 (1955), S. 411-416 
    Details
    ISSN: 1573-5060
    Keywords: Agrobacterium ; Brassica napus ; CaMV 35S promoter ; mas promoter ; gene expression ; risk assessment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Gene fusions between the β-glucuronidase (GUS) reporter gene and the promoters of the cauliflower mosaic virus 35S RNA transcript (CaMV 35S) and the mannopine synthase (mas) genes were introduced into rapeseed varieties via Agrobacterium-mediated transformation. Fluorometric assay of β-glucuronidase activity indicated different expression patterns for the two promoters. In seedlings, the CaMV 35S promoter had maximum activity in the primary roots, while the mas promoter was most active in the cotyledons. Etiolated seedlings cultured in the dark showed reduced activity of the mas promoter. Before vernalization at the rosette stage, both promoters were more active in older plant parts than in younger ones. At this stage the highest activity was recorded in cotyledons. After the plants had bolted reduced promoter function was detected in the upper parts of the transformed plants. Both promoters were found to be functional in the majority of the studied organs of transgenic rapeseed plants, but the promoter activity varied considerably between the organs at different developmental stages. The ability of pollen to transfer the introduced genes to other varieties and related species (e.g. Brassica napus and Diplotaxus muralis) by cross-pollination was studied in greenhouse experiments, and field trials were carried out to estimate the distance for biologically-relevant gene dispersal. In artificial crossing, the introduced marker gene was transferable into other varieties of Brassica napus. In field trials, at a distance of 1 metre from the source of transgenic plants, the frequency of an outcrossing event was relatively high (10-3). Resistant individuals were found at 16 and 32 metres from the transgenic pollen donors, but the frequency of an outcrossing event dropped to 10-5.
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    URL: http://dx.doi.org/10.1007/BF00023974
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  • 27
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    Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants (1955)
    Springer
    Euphytica 85 (1955), S. 81-88 
    Details
    ISSN: 1573-5060
    Keywords: gene transfer ; Hordeum vulgare ; neomycin phosphotransferase II ; particle bombardment ; transgenic barley
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for β-glucuronidase (GUS). Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T0). This chimeric plant passed the transferred nptII gene to its T1 progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T0 spikes and 15 T1 offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T2 and T3 plants were produced by isolating embryos from green grains of transgenic T1 and T2 plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T2 offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed. Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.
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    URL: http://dx.doi.org/10.1007/BF00023933
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    Transfer of disease resistance within the genus Brassica through asymmetric somatic hybridization (1955)
    Springer
    Euphytica 85 (1955), S. 247-253 
    Details
    ISSN: 1573-5060
    Keywords: asymmetric somatic hybridization ; Brassica napus ; Brassica nigra ; disease resistance transfer ; dot blot analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Asymmetric somatic hybrid plants between Brassica napus L. (oilseed rape genome AACC) and a transgenic line of Brassica nigra L. Koch (black mustard genome BB) were tested for their resistance against rapeseed pathogens Phoma lingam (black leg disease) and Plasmodiophora brassicae (club root disease). The transgenic B. nigra line used (hygromycin-resistant, donor) is highly resistant to both fungi, whereas B. napus (recipient) is highly susceptible. The asymmetric somatic hybrids were produced using the donor-recipient fusion method (with X-irradiation of donor protoplasts) reported by Zelcer et al. (1978) for the production of cybrids. Using hygromycin-B for selection, a total of 332 hybrid calli were obtained. Regenerants, resistant or susceptible to both diseases, were selected. Many hybrids expressed resistance to only one pathogen. Dot blot experiments showed that the asymmetric hybrid plants contained varying amounts of the donor genomic DNA. Furthermore, a correlation was detected between the radiation dose and the degree of donor DNA elimination.
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    URL: http://dx.doi.org/10.1007/BF00023953
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  • 29
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    Degradation of oxalic acid by transgenic oilseed rape plants expressing oxalate oxidase (1955)
    Springer
    Euphytica 85 (1955), S. 169-172 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; disease tolerance ; oxalic acid ; oxalate oxidase ; Sclerotinia sclerotiorum ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Oxalic acid is thought to have a primary role in the pathogenicity of several plant pathogens, notably Sclerotinia selerotiorum. A gene coding for the enzyme oxalate oxidase was isolated from barley roots and introduced into oilseed rape as a means of degrading oxalic acid in vivo. This report describes the production of several transgenic plants of oilseed rape and the characterisation of these plants by Southern, Western and enzyme activity assays. Plants were shown to contain an active oxalate oxidase enzyme and were tolerant of exogenously supplied oxalic acid.
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    URL: http://dx.doi.org/10.1007/BF00023945
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  • 30
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    Modification of oilseed rape to produce oils for industrial use by means of applied tissue culture methodology (1955)
    Springer
    Euphytica 85 (1955), S. 323-327 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napus ; fatty acids ; gas chromatography ; Lunaria annua ; protoplast regeneration ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A programme of research was designed to investigate methods for the modification of the fatty acid profiles of high performance lines of oilseed rape (Brassica napus L.) in an attempt to produce lines with enhanced levels of industrially useful fatty acids. The methodology employed to achieve these objectives was based on the exploitation of somaclonal or protoclonal variation, and targeted somatic hybridization using wild cruciferous germplasm as fusion partners. A range of somaclonal lines was produced from shoot regeneration protocols. These lines underwent replicated, randomised glasshouse trials for morphological assessment followed by gas chromatographic analysis to monitor any changes in fatty acid profile. It was found that a small number of lines exhibited potentially useful changes in oleic acid and polyunsaturated fatty acid content. Protoplast regeneration and electrofusion protocols for a range of winter oilseed rape lines were developed, and methods for the isolation and fusion of protoplasts of the wild crucifer Lunaria annua (chosen for its high nervonic acid content) established.
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    URL: http://dx.doi.org/10.1007/BF00023962
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    The sulphur-rich Brazil nut 2S albumin is specifically formed in transgenic seeds of the grain legume Vicia narbonensis (1955)
    Springer
    Euphytica 85 (1955), S. 181-192 
    Details
    ISSN: 1573-5060
    Keywords: Vicia narbonensis ; gene transfer ; gene expression ; seeds ; 2S albumin ; methionine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Epicotyl explants were co-cultivated with Agrobacterium tumefaciens EHA101 to transfer a chimeric 2S albumin gene construct carried in the binary Ti plasmid vectors pGSGLUC1 or pGA472 into the grain legume Vicia narbonensis. This gene encoding the sulphur-rich Brazil nut albumin was under the control of either the CaMV 35S promoter which permits gene expression in all organs, or the Vicia faba legumin B4 promoter which elicits seed-specific gene expression. After callus formation and selection for kanamycin resistance, somatic embryos were induced which, in the case of transformation with the vector pGSGLUC1, were screened for GUS activity. Embryos that produced GUS were in addition analysed for 2S albumin formation. Selected transgenic embryos were cloned by multiple shoot regeneration. Rooted and fertile plants were obtained by grafting transgenic shoots on the appropriate seedlings. R1 and R2 generations were raised and analysed for GUS as well as 2S albumin gene expression. Expression of the 35S promoter/2S albumin gene fusion took place in all organs of the transgenic plants including the cotyledons of seeds, whereas seed-specific gene expression was found in transformants with the legumin promoter/2S albumin gene fusion. The 2S albumin accumulated in the 2S protein fraction of transgenic seeds and its primary translation product was processed into the 9 and 3 kDa polypeptide chains. The foreign protein was localised in the protein bodies of the grain legume. Analysis of the R2 plants indicated Mendelian inheritance of the 2S albumin gene. In homozygous V. narbonensis plants the amounts of 2S albumin were twice that present in the corresponding heterozygous plants. Whereas only low level formation of the foreign protein was achieved if the gene was under the control of the 35S promoter, approximately 3.0% of the soluble seed protein was 2S albumin if seed-specific gene expression was directed by the legumin B4 promoter. Some of these transformants exhibited a three-fold increase in the methionine content of the salt-soluble protein fraction extracted from seeds.
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    URL: http://dx.doi.org/10.1007/BF00023947
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    Strategies for variety-independent genetic transformation of important cereals, legumes and woody species utilizing particle bombardment (1955)
    Springer
    Euphytica 85 (1955), S. 13-27 
    Details
    ISSN: 1573-5060
    Keywords: gene transfer ; crop species ; particle bombardment ; transgenic plants ; cereals ; legumes ; woody plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The limiting component in the creation of transgenic crops has been the lack of effective means to introduce foreign genes into elite germplasm. However, the development of novel direct DNA transfer methodology, by-passing limitations imposed by Agrobacterium-host specificity and cell culture constraints, has allowed the engineering of almost all major crops, including formerly recalcitrant cereals, legumes and woody species. The creation of transgenic rice, wheat, maize, barley, oat, soybean, phaseolus, peanut, poplar, spruce, cotton and others, in an efficient and in some cases, variety-independent fashion, is a significant step towards the routine application of recombinant DNA methodology to the improvement of most important agronomic crops. In this review we will focus on key elements and advantages of particle bombardment technology in order to evaluate its impact on the accelerated commercialization of products based on agricultural biotechnology and its utility in studying basic plant developmental processes and function through transgenesis. Fundamental differences between conventional gene transfer methods, utilizing Agrobacterium vectors or protoplast/suspension cultures, and particle bombardment will be discussed in depth.
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    URL: http://dx.doi.org/10.1007/BF00023926
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