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  • 1
    Electronic Resource
    Electronic Resource
    Identification of a Novel Plant Virus Promoter using a Potyvirus Infectious Clone (2000)
    Springer
    Virus genes 20 (2000), S. 11-17 
    Details
    ISSN: 1572-994X
    Keywords: plant viral promoter ; caulimovirus ; strawberry vein banding virus ; zucchini yellow mosaic virus ; particle bombardment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A putative promoter from the strawberry vein banding caulimovirus (SVBV) genome was identified by its ability to drive infection with full-length cDNA of the zucchini yellow mosaic RNA potyvirus (ZYMV). A high rate of infection was obtained with the cDNA under control of the SVBV promoter using particle bombardment technology. The SVBV promoter shows 60% homology to the cauliflower mosaic virus 35S promoter in the domain spanning the conserved motifs of CCACT (at −83) and the TATA box (at −31), to the transcription start. The 3′-end one-third of the putative promoter (328 bp) was sufficient to invoke full infectivity with the ZYMV clone, and drove transient reporter gene expression in Solanaceae and Cucurbitaceae transformed with a binary plant transformation vector. Stable expression of a reporter gene (GUS) under control of the truncated SVBV promoter was shown in transformed tobacco shoots in roots, leaves and stems.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008199805099
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  • 2
    Electronic Resource
    Electronic Resource
    The effect of DNA/gold particle preparation technique, and particle bombardment device, on the transformation of barley (Hordeum vulgare) (2000)
    Springer
    Euphytica 111 (2000), S. 67-76 
    Details
    ISSN: 1573-5060
    Keywords: Barley ; Hordeum vulgare ; transformation ; particle bombardment ; particle gun
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Immature embryos of the spring barley variety GoldenPromise, were bombarded with three different particledelivery systems and both transient and stabletransformation examined. In addition, a range oftechniques for the preparation of the DNA coated goldparticles was examined. Fertile transgenic barleyplants were obtained using three particle preparationtechniques which differed in the amount of gold andDNA used for each bombardment. However, only one ofthe particle delivery systems, the PDS 1000/He device,appeared to be effective in yielding transformedbarley plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003700300235
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  • 3
    Electronic Resource
    Electronic Resource
    Direct transformation and plant regeneration of the haploid liverwort Marchantia polymorpha L. (2000)
    Springer
    Transgenic research 9 (2000), S. 179-185 
    Details
    ISSN: 1573-9368
    Keywords: direct transformation ; haploid plant ; Marchantia polymorpha ; particle bombardment ; plant regeneration ; stable integration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Thalli of the haploid liverwort Marchantia polymorpha were successfully used for direct particle bombardment with plasmid pMT, which carries a hygromycin phosphotransferase gene (hpt) controlled by the CaMV 35S promoter and the NOS polyadenylation region. Hygromycin-resistant cell masses arose from the thallus surface and developed directly into hygromycin-resistant thalli. Southern blot analyses indicated that these thalli carried at least 1–4 copies of the hpt gene, which were stably transmitted to their asexual thallus progenies via gemma propagation for three generations. This transformation and direct plant regeneration protocol is expected to be a valuable tool for the molecular analysis of this lower land plant.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008963410465
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  • 4
    Electronic Resource
    Electronic Resource
    Overexpression of The Calcium-Dependent Protein Kinase OsCDPK2 in Transgenic Rice is Repressed by Light in Leaves and Disrupts Seed Development (2000)
    Springer
    Transgenic research 9 (2000), S. 453-462 
    Details
    ISSN: 1573-9368
    Keywords: calcium-dependent ; light regulation ; particle bombardment ; protein kinase ; seed development ; transgenic rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Independent transgenic rice lines overexpressing the rice CDPK isoform OsCDPK2 were generated by particle bombardment. High levels of OsCDPK2 were detected in leaves removed from etiolated plants, as well as in stems and flowers. However, there was no overexpression in green leaves that had been exposed to light, confirming that OsCDPK2 protein stability was subject to light regulation. The morphological phenotype of transgenic plants producing high levels of recombinant OsCDPK2 was normal until the onset of seed development. Flowers developed normally, producing well-shaped ovaries and stigmas, and mature anthers filled with pollen grains. However, seed formation in these plants was strongly inhibited, with only 3–7% of the flowers producing seeds. Seed development was arrested at an early stage. We discuss these data with respect to the possible requirement for specific CDPK isoforms during rice seed 4.4ptdevelopment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1026555021606
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  • 5
    Electronic Resource
    Electronic Resource
    Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns (2000)
    Springer
    Transgenic research 9 (2000), S. 11-19 
    Details
    ISSN: 1573-9368
    Keywords: particle bombardment ; minimal cassette ; cotransformation ; transgenic rice ; integration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Whole plasmids are used in both Agrobacterium-mediated transformation and direct DNA transfer, generally leading to the integration of vector backbone sequences into the host genome along with the transgene(s). This is undesirable, as vector backbone sequences often have negative effects on transgene or endogenous gene expression, and can promote transgene rearrangements. We, therefore, bombarded rice tissue with two constructs: a plasmid containing the bar gene, and a linear DNA fragment isolated from the same plasmid, corresponding to the minimal bar gene expression cassette (promoter, open reading frame and terminator). We recovered phosphinothricin-resistant plants from both experiments, showing that the selectable marker was efficiently expressed. Transformation with such constructs resulted in predominantly 'simple' integration events (one or two bands on Southern blots), producing low-copy-number transgenic plants with a low frequency of transgene rearrangements. Conversely, transformation with supercoiled or linearized whole plasmids generated plants with 'complex' integration patterns, that is, higher copy numbers and frequent transgene rearrangements. We monitored transgenic lines through to the R4 generation and observed no silencing in plants carrying minimal constructs. We also carried out experiments in which rice tissue was simultaneously bombarded with minimal linear hpt and gusA cassettes. We observed robust GUS activity in hygromycin-resistant plants, confirming co-expression of the selectable and nonselectable markers. Furthermore, the efficiency of cotransformation using minimal constructs was the same as that using supercoiled plasmid cointegrate vectors.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008993730505
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  • 6
    Electronic Resource
    Electronic Resource
    Transformation of the tropane alkaloid-producing medicinal plant Hyoscyamus muticus by particle bombardment (2000)
    Springer
    Transgenic research 9 (2000), S. 163-168 
    Details
    ISSN: 1573-9368
    Keywords: Hyoscyamus muticus ; particle bombardment ; transformation ; tropane alkaloids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report an efficient whole plant transformation system for Hyoscyamus muticus, an important medicinal plant of the Solanaceous family. We developed a system using a plasmid carrying the nptII and gusA genes, which was delivered into leaf explants by particle bombardment. Ten percent of bombarded leaf explants formed kanamycin-resistant callus, from which putative transgenic plants were recovered. The nptII gene conferring kanamycin resistance was found to be incorporated into the genome of all transgenic plants screened. Over 50% of the kanamycin resistant plants showed strong expression of the non-selected gusA gene. The majority of transgenic plants reached maturity, could be self pollinated, and produced fertile seed. A simple and efficient whole plant transformation system for this medicinal plant is an important step in furthering our understanding of tropane alkaloid production in plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008912330067
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  • 7
    Electronic Resource
    Electronic Resource
    Cereal crops as viable production and storage systems for pharmaceutical scFv antibodies (2000)
    Springer
    Plant molecular biology 42 (2000), S. 583-590 
    Details
    ISSN: 1573-5028
    Keywords: transgenic cereals ; molecular pharming ; antibodies ; carcinoembryonic antigen ; single-chain Fv fragment ; particle bombardment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This report describes the stable expression of a medically important antibody in the staple cereal crops rice and wheat. We successfully expressed a single-chain Fv antibody (ScFvT84.66) against carcinoembryonic antigen (CEA), a well characterized tumor-associated marker antigen. scFv constructs were engineered for recombinant antibody targeting to the plant cell apoplast and ER. Up to 30 μg/g of functional recombinant antibody was detected in the leaves and seeds of wheat and rice. We confirmed that transgenic dry seeds could be stored for at least five months at room temperature, without significant loss of the amount or activity of scFvT84.66. Our results represent the first transition from model plant expression systems, such as tobacco and Arabidopsis, to widely cultivated cereal crops, such as rice and wheat, for expression of an antibody molecule that has already shown efficacy in clinical applications. Thus, we have established that molecular pharming in cereals can be a viable production system for such high-value pharmaceutical macromolecules. Our findings provide a strong foundation for exploiting alternative uses of cereal crops both in industrialized and developing countries.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006301519427
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  • 8
    Electronic Resource
    Electronic Resource
    Enhanced insect resistance in Thai rice varieties generated by particle bombardment (2000)
    Springer
    Molecular breeding 6 (2000), S. 391-399 
    Details
    ISSN: 1572-9788
    Keywords: cotransformation ; gna gene ; insect resistance ; Oryza sativa L. ; particle bombardment ; transgenic Thai rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We used particle bombardment to transform two elite Thai rice varieties, Khao Dawk Mali 105 (KDML105) and Supanburi 60 (SP60), with the snowdrop lectin gene gna (Galanthus nivalis agglutinin). This gene confers resistance to sap-sucking insects such as the brown planthopper (BPH; Nilaparvata lugens), which is one of the most damaging pests of rice. Traditionally, KDML105 and SP60 have been regarded as recalcitrant to transformation, and this is the first account of successful gene transfer to these varieties. By molecular analysis, we confirmed the recovery of over thirty gna-transgenic lines. GNA protein expression was characterised by western blot analysis, and we achieved expression levels of up to 0.25% total soluble protein. GNA-producing R1 transgenic plants were significantly more resistant to BPH than control plants (P〈0.0001), with 37% and 42% reduction in nymphal survival for constitutive and phloem-specific expression, respectively. Transferring the gna gene to these superior rice varieties thus represents a major step forward for crop improvement in Thailand, and should help to reduce the damage caused by rice pests, and hence increase yields for this vital domestic and export market.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009633703157
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