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  • Populus  (4)
  • Biolistics-GUS-355-Conifer  (1)
  • 1
    ISSN: 1432-203X
    Keywords: Biolistics-GUS-355-Conifer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plasmid DNA containing the reporter gene uidA encoding β-glucuronidase (GUS), driven by the cauliflower mosaic virus 35S promoter, was introduced on high-velocity microprojectiles into cultured cotyledons of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco]. Transient gene expression was measured by counting the number of distinct loci of GUS activity per cotyledon. Contrary to published results on angiosperms, repeated bombardments did not increase expression in Douglas-fir. Expression varied significantly among cotyledons from different seedlings. The amount of time between DNA delivery and treatment of cotyledons with auxins and cytokinins strongly affected GUS expression. The optimal cytokinin pretreatment produced an average of 20 loci per cotyledon. In several experiments, more than 95% of the treated cotyledons exhibited at least some transient expression. Expression remained constant up to three days following DNA delivery into cotyledons.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 59-71 
    ISSN: 1573-5044
    Keywords: antibiotics ; cocultivation ; organogenesis ; Populus ; tissue culture ; thidiazuron
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method for Agrobacterium-mediated transformation of hybrid poplar (Populus alba x P. grandidentata cv. ‘Crandon’) suspension cultures and regeneration of transformed plants is described. Transformants were recovered when suspension cultures were inoculated with Agrobacterium tumefaciens at a density of 107 colony-forming units ml-1, cocultivated for 48 h, and plated to cellulose acetate filters on Woody Plant Medium containing 4.5 μM 2,4-dichlorophenoxyacetic acid and 250 mg l-1 cefotaxime. Levels of cefotaxime greater than 250 mg l-1 were unnecessary for control of residual bacteria and inhibited callus growth. Transgenic plants were regenerated by culturing the transformed callus on media containing 0.11 to 27 μM thidiazuron. In contrast to thidiazuron, N6-benzyladenine had a negative effect on shoot regeneration; the callus became necrotic when we attempted to induce shoots with concentrations of 1.1 to 8.9 μM, and growth was inhibited when concentrations of 0.11 or 0.22 μM were used to regenerate callus from suspension cultures. Following cocultivation of poplar suspension cultures, we recovered transgenic plants containing the maize transposon Ac, and callus containing an insect toxin gene from Bacillus thuringiensis.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5028
    Keywords: AGAMOUS ; cottonwoods ; dioecy ; floral development ; MADS-box ; Populus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To investigate the homeotic systems underlying floral development in a dioecious tree, and to provide tools for the manipulation of floral development, we have isolated two Populus trichocarpa genes, PTAG1 and PTAG2, homologous to the Arabidopsis floral homeotic gene AGAMOUS (AG). PTAG1 and PTAG2 are located on separate linkage groups, but their non-coding regions are highly similar, consistent with a phylogenetically recent duplication. Intron/exon structure is conserved in relation to AG and the Antirrhinum AG orthologue, PLENA (PLE), and low-stringency Southern analysis demonstrated the absence of additional genes in the poplar genome with significant PTAG1/2 homology. PTAG1 and PTAG2 exhibit an AG-like floral expression pattern, and phylogenetic analysis of the AG subfamily strongly supports evolutionary orthology to C-class organ identity genes. The high degree of similarity shared by PTAG1 and PTAG2 in both sequence (89% amino acid identity) and expression indicates that they are unlikely to be functionally associated with specification of tree gender. Unexpectedly, PTAG transcripts were consistently detected in vegetative tissues.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1572-9788
    Keywords: flowering ; floral homeotic genes ; ablation ; cosuppression ; antisense ; transgenes ; Populus ; conifer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Containment of transgenes inserted into genetically engineered forest trees will probably be necessary before most commercial uses are possible. This is a consequence of (1) high rates of gene dispersal by pollen and seed, (2) proximity of engineered trees in plantations to natural or feral stands of interfertile species, and (3) potentially undesirable ecological effects if certain transgenes become widely dispersed. In addition to gene containment, engineering of complete or male sterility may stimulate faster wood production, reduce production of allergenic pollen, and facilitate hybrid breeding. We review the regulatory and ecological rationale for engineering sterility, potentially useful floral genes, strategies for creating sterility-causing transgenes, and problems peculiar to engineering sterility in forest trees. Each of the two primary options — ablating floral tissuesvia floral promoter-cytotoxin fusions, and disrupting expression of essential floral genes by various methods of gene suppression — has advantages and disadvantages. Because promoters from structural and enzymatic floral-specific genes often work well in heterologous species, ablation methods based on these genes probably will not require cloning of homologs from angiosperm trees. Methods that inhibit gene expression will require cloning of tree genes and may be more prone to epigenetic variability, but should allow assay of transgene efficacy in seedlings. Practical constraints include the requirement for vegetative propagation if complete sterility is engineered and the need for highly stable forms of sterility in long-lived trees. The latter may require suppression of more than one floral gene or employment of more than one genetic mechanism for sterility.
    Type of Medium: Electronic Resource
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  • 5
    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
    Type of Medium: Electronic Resource
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