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  • Ultrastructure  (52)
  • Transformation  (36)
  • Springer  (88)
  • 1995-1999  (42)
  • 1980-1984  (46)
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  • Springer  (88)
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  • 1
    ISSN: 1432-203X
    Keywords: Key words Forage grasses ; Dactylis glomerata L. ; Orchardgrass ; Transformation ; Microprojectile bombardment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Young leaf tissue of orchardgrass (Dactylis glomerata L.) was placed on Schenk and Hildebrandt medium containing 30 µM dicamba. Microprojectiles coated with DNA containing the selectable bar gene (Basta® tolerance) and the reporter gene uidA coding for β-glucuronidase (GUS), both driven by the maize ubiquitin promoter (Ubi1), were propelled into the tissue with a particle inflow gun. Transient GUS expression was observed as blue spots of various sizes on leaf segments. Somatic embryos staining entirely blue were also produced, and embryos germinated on medium containing 3.0 mg 1–1 bialaphos. Leaves of 67 putative transformed plants were painted with 0.1% Basta. Ten showed no reaction, and 6 showed only a localized response. Cultured leaf segments from tolerant plants also produced somatic embryos that expressed GUS. The genetic transformation was confirmed by Southern blot hybridization and PCR analyses of T0 plants and by PCR analyses of somatic embryos produced from T0 plants.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 107 (1981), S. 85-107 
    ISSN: 1615-6102
    Keywords: Male cytoplasmic inheritance ; Plumbago ; Pollen grain ; Pollen tube ; Sperm ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Male gametes ofPlumbago zeylanica were examined in pollen grains and tubes using light and electron microscopy of chemically and physically fixed tissues, and Nomarski interference microscopy of isolated, living sperm cells. Male gametes are elongate, spindleshaped cells containing a nucleus, mitochondria, ER, ribosomes, vesicles, dictyosomes, probable microfilaments, and a variable number of plastids. In mature pollen grains ofP. zeylanica, the two sperm cells are directly linked; they share a transverse cell wall with plasmodesmata and are enclosed together by the inner vegetative cell plasma membrane. One of these two sperms is also associated with the vegetative nucleus as a consistent feature of pollen grain organization. The basis of this association appears to be a long, narrow projection of the sperm cell (averaging 〈 1 μm wide and about 30 μm long) which wraps around the periphery of the vegetative nucleus and occupies embayments of that nucleus. This association is maintained throughout pollen tube growth but becomes less extensive near the completion of tube growth and is severed following tube discharge. The consistent occurrence of the sperm-vegetative nucleus association in pollen grains, tubes and isolated pollen cytoplasm suggests that the two structures may be directly connected, but attempts to visualize this type of connection were unsuccessful. Possibly, the entwining nature and extent of complementary interfaces between vegetative nucleus and sperm may have a role in stabilizing their association. Functionally, the two sperms and vegetative nucleus appear to travel as a linked unit within the pollen tube, possibly increasing the effectiveness of gamete delivery and helping to ensure nearly simultaneous transmission of sperms into the receptive megagametophyte.
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  • 3
    ISSN: 1432-203X
    Keywords: Key wordsPinus radiata ; Transformation ; Biolistic ; Embryogenic tissue
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A biolistic particle delivery system was used to genetically transform embryogenic tissue of Pinus radiata. The introduced DNA contained a uidA reporter gene under the control of either the tandem CaMV 35S or the artificial Emu promoter, and the npt II selectable marker controlled by the CaMV 35S promoter. The average number of stable, geneticin-resistant lines recovered was 0.5 per 200 mg fresh weight bombarded tissue. Expression of the uidA reporter gene was detected histochemically and fluorimetrically in transformed embryogenic tissue and in derived mature somatic embryos and regenerated plants. The integration of uidA and npt II genes into the Pinus radiata genome was demonstrated using PCR amplification of the inserts and Southern hybridisation analysis. The expression of both genes in transformed tissue was confirmed by Northern hybridisation analysis. More than 150 transgenic Pinus radiata plants were produced from 20 independent transformation experiments with four different embryogenic clones.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 133 (1982), S. 11-19 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Ultrastructure ; Mastigocladus laminosus ; Fischerella ; True branching
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The morphology and ultrastructure of the thermophilic cyanobacteriumMastigocladus laminosus were examined by scanning and transmission electron microscopy. Mature cultures consisted of relatively old, wide filaments that branched frequently to form younger, thinner filaments. The cells of the younger filaments had a consistently cylindrical morphology, while those of older filaments were rounded and pleomorphic. The internal ultrastructure of the cells depended somewhat on their age. As young cells became larger and wider, their thylakoids underwent slight rearrangement and spread out toward the center of the cytoplasm. Polyphosphate bodies, carboxysomes (polyhedral bodies), and lipid-body-like structures increased in number as the cells aged, but ribosomes and cyanophycin granules were depleted. Cell division involved septum formation followed by ingrowth of the outer membrane and sheath. Cells in older filaments were separated from each other by a complete layer of sheath material. Septum formation in older cells was also seen to occur parallel to the long axis of the filament, thereby confirming that true branching took place.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Archives of microbiology 130 (1981), S. 204-212 
    ISSN: 1432-072X
    Keywords: Agmenellum quadruplicatum ; Nitrogen starvation ; Ultrastructure ; PATO poststain ; Cyanobacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of nitrogen limitation on the ultrastructure of the unicellular cyanobacterium, Agmenellum quadruplicatum, were studied by thin sectioning transmission electron microscopy. Nitrogen became limiting for growth 14–15 h after transfer to nitrogen-limiting medium, but cultures retained full viability for at least 45 h. The c-phycocyanin: chlorophyll a ratio and cellular nitrogen content of the culture dropped rapidly after 14–15 h, as a progressive deterioration of major cell structures took place. Phycobilisomes were degraded first, followed by ribosomes and, then, thylakoid membranes. These structures were virtually depleted from the cells within 26 h. Intracellular polysaccharide accumulated in place of the normal cell structures throughout this period. Nitrogen limitation did not affect polyphosphate bodies, carboxysomes, lipid granules, the cell envelope, or the extra-cellular glycocalyx. All of the ultrastructural changes resulting from nitrogen limitation were reversed upon addition of nitrate to a starved culture. Most cell structures were restored within 3 h, and restoration was complete within 9 h.
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  • 6
    ISSN: 1432-2242
    Keywords: Maize ; Transformation ; Lysine ; Dihydrodipicolinate synthase ; Aspartate kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Lysine is one of the nutritionally limiting amino acids in food and feed products made from maize (Zea mays L.). Two enzymes in the lysine biosynthesis pathway, aspartate kinase (AK) and dihydrodipicolinate synthase (DHPS), have primary roles in regulating the level of lysine accumulation in plant cells because both enzymes are feedback-inhibited by lysine. An isolated cDNA clone for maize DHPS was modified to encode a DHPS much less sensitive to lysine inhibition. The altered DHPS cDNA was transformed into maize cell suspension cultures to determine the effect on DHPS activity and lysine accumulation. Partially purified DHPS (wildtype plus mutant) from transformed cultures was less sensitive to lysine inhibition than wild-type DHPS from nontransformed cultures. Transformed cultures had cellular free lysine levels as much as four times higher than those of nontransformed controls. Thus, we have shown that reducing the feedback inhibition of DHPS by lysine can lead to increased lysine accumulation in maize cells. Increasing the capacity for lysine synthesis may be an important step in improving the nutritional quality of food and feed products made from maize.
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  • 7
    ISSN: 1432-2242
    Keywords: Key wordsTriticum aestivum ; Transformation ; Microprojectile bombardment ; Chitinase gene ; bar gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Our long-term goal is to control wheat diseases through the enhancement of host plant resistance. The constitutive expression of plant defense genes to control fungal diseases can be engineered by genetic transformation. Our experimental strategy was to biolistically transform wheat with a vector DNA containing a rice chitinase gene under the control of the CaMV 35 S promoter and the bar gene under control of the ubiquitin promoter as a selectable marker. Immature embryos of wheat cv ‘Bobwhite’ were bombarded with plasmid pAHG11 containing the rice chitinase gene chi11 and the bar gene. The embryos were subcultured on MS2 medium containing the herbicide bialaphos. Calli were then transferred to a regeneration medium, also containing bialaphos. Seventeen herbicide-resistant putative transformants (T0) were selected after spraying with 0.2% Liberty, of which 16 showed bar gene expression as determined by the phosphinothricin acetyltransferase (PAT) assay. Of the 17 plants, 12 showed the expected 35-kDa rice chitinase as revealed by Western blot analysis. The majority of transgenic plants were morphologically normal and self-fertile. The integration, inheritance and expression of the chi11 and bar genes were confirmed by Southern hybridization, PAT and Western blot analysis of T0 and T1 transgenic plants. Mendelian segregation of herbicide resistance was observed in some T1 progenies. Interestingly, a majority of the T1 progeny had very little or no chitinase expression even though the chitinase transgene was intact. Because PAT gene expression under control of the ubiquitin promoter was unaffected, we conclude that the CaMV 35 S promoter is selectively inactivated in T1 transgenic wheat plants.
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  • 8
    ISSN: 1432-0878
    Keywords: Ecdysteroids ; Moulting hormones ; Xyleborus ferrugineus pupae ; Ultrastructure ; Prothoracic gland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The prothoracic glands of female pupae of Xyleborus ferrugineus at three ages (0-h, 48-h and 72-h-old) were examined for ultrastructural changes that correlate with high titers of ecdysteroids in the entire pupa. In all three ages, the prothoracic gland cells appear compact with a prominent nucleus. Lysosome-like structures with concentrically oriented internal membrane are observed in both 0-h and 72-h-old gland cells. Mitochondria are abundant in the glands of both ages, and rough endoplasmic reticulum is relatively sparse. Numerous microtubules are present in the prothoracic gland cells of 48-h-old female pupae that contain relatively high titers of ecdysteroids. Rough endoplasmic reticulum also is especially abundant at this age of the pupa. Ecdysone titers, estimated by the radioimmunoassay method (RIA), were 161.04±22.61 pg/mg, 704.25±69.02 pg/mg and 298.72±26.80pg/mg body weight in 0-h, 48-h and 72-h-old female pupae, respectively.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 251 (1996), S. 23-30 
    ISSN: 1617-4623
    Keywords: Chlamydomonas ; Transformation ; Dominant marker ; ble
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A chimeric gene composed of the coding sequence of theble gene fromStreptoalloteichus hindustanus fused to the 5′ and 3′ untranslated regions of theChlamydomonas reinhardtii nuclear geneRBCS2 has been constructed. Introduction of this chimeric gene into the nuclear genome ofC. reinhardtii by co-transformation with theARG7 marker yields Arg+ transformants of which approximately 80% possess theble gene. Of these co-transformants, approximately 3% display a phleomycin-resistant (PmR) phenotype. Western blot analysis using antibodies against theble gene product confirms the presence of the protein in the PmR transformants and genetic analysis demonstrates the co-segregation of theble gene with the phenotype in progeny arising from the mating of a PmR transformant to wild-type strains. Direct selection of PmR transformants was achieved by allowing an 18-h period for recovery and growth of transformed cells prior to selection. This work represents the first demonstration of stable expression and inheritance of a foreign gene in the nuclear genome ofC. reinhardtii and provides a useful dominant marker for nuclear transformation.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 116 (1983), S. 1-13 
    ISSN: 1615-6102
    Keywords: Cuticle ; Peristomatal transpiration ; Stomata ; Ultrastructure ; Funaria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cuticle and pore development in the guard cells ofFunaria were investigated with the electron microscope. Pore cuticle formation is simultaneous with the creation of the pore itself. The morphology of the pore cuticle is unlike that of any cuticle described in the literature. It has many lamellae which are penetrated by electron dense fibrils. Three different cuticular morphologies exist from the pore to the subsidiary cell walls. The cuticles on the pore and outer walls contain fibrils that sometimes reach to the surface. The subsidiary cell cuticle lacks fibrils altogether. It is hypothesized that (1) cuticularization of the middle lamella contributes to ventral wall separation and (2) differences in extent of cuticular fibrils are related to greater water loss from stomata than from subsidiary cells (peristomatal transpiration).
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