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  • DAPI  (5)
  • Chloroplast nuclei  (3)
  • Amyloplast  (2)
  • Biparental inheritance  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Amyloplast formation in cultured tobacco cells. III Determination of the timing of gene expression necessary for starch accumulation (1999)
    Springer
    Plant cell reports 18 (1999), S. 589-594 
    Details
    ISSN: 1432-203X
    Keywords: Key words ADP-glucose starch glycosyl transferase ; Amyloplast ; BY-2 ; Nicotiana tabacum ; Transcription/translation inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When BY-2 cultured tobacco (Nicotiana tabacum L.) cells were transferred to auxin-depleted culture medium containing cytokinin (benzyladenine, 1 mg/l), the starch content per cell started increasing from 18 h of culture and amyloplasts had formed by 48 h. Pulse-treatment of the cells with actinomycin D and cycloheximide for the first 12 h (or longer) of culture significantly decreased the cellular starch content after 48 h, whereas the starch content did not decrease significantly when the cells were released from the inhibition within 6 h. This suggests that nuclear gene expression necessary for amyloplast formation begins 6–12 h after the transfer. Immunoblotting analysis of the accumulation of ADP-glucose starch glycosyl transferase (starch synthase) supported this inference.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050627
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  • 2
    Electronic Resource
    Electronic Resource
    Cytoplasmic DNA apportionment and plastid differentiation during male gametophyte development in Pelargonium zonale (1994)
    Springer
    Sexual plant reproduction 7 (1994), S. 51-56 
    Details
    ISSN: 1432-2145
    Keywords: Cytoplasmic DNA apportionment ; Biparental inheritance ; Plastid differentiation ; Male gametophyte ; Pelargonium zonale
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the male gametophyte of Pelargonium zonale, generative and sperm cells contain cytoplasmic DNA in high density compared to vegetative cells. Cytoplasmic DNA was examined using the DNA fluorochrome DAPI (4'6-diamidino-2-phenylindole) and observed with epifluorescence and electron microscopy. The microspore cell contains a prominent central vacuole before mitosis; mitochondria and plastids are randomly distributed throughout the cytoplasm. Following the first pollen grain mitosis, neither the vegetative cell nor the early generative cell display a distributional difference in cytoplasmic DNA, nor is there in organelle content at this stage. During the maturation of the male gametophyte, however, a significant discrepancy in plastid abundance develops. Plastids in the generative cell return to proplastids and do not contain large starch grains, while those in the vegetative cell develop starch grains and differentiate into large amyloplasts. Plastid nucleoids in generative and sperm cells in a mature male gametophyte are easily discriminated after DAPI staining due to their compactness, while those in vegetative cells stained only weakly. The utility of the hydrophilic, non-autofluorescent resin Technovit 7100 in observing DAPI fluorescence is also demonstrated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00241887
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  • 3
    Electronic Resource
    Electronic Resource
    Behavior of leucoplast nucleoids in the epidermal cell of onion (Allium cepa) bulb (1982)
    Springer
    Protoplasma 110 (1982), S. 177-184 
    Details
    ISSN: 1615-6102
    Keywords: Leucoplast division cycle ; Leucoplast nucleoid ; DAPI ; Allium cepa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The behavior of nucleoids during the leucoplast division cycle in the epidermis of onion (Allium cepa) bulbs was investigated using DNA-specific fluorochrome 4'6-diamidino-2-phenylindole (DAPI) staining. The leucoplast was morphologically amoeboid and continuously changed its shape. A dumbbell-shaped leucoplast divided into two spherical daughter ones by constriction in the middle region of the body. Leucoplasts contained 4–10 mostly spherical, oval, partly rodand dumbbell-shaped nucleoids which were dispersed within the bodies. The proportion of one DNA molecule of a T4 phage particle to the small leucoplast nucleoid in the grain density of negative film was 1 to 0.91. Comparison of the present result and another groups' biochemical results suggested that a small leucoplast nucleoid contains one DNA molecule. The dumbbell-shaped leucoplast probably before division contained about twice as many nucleoids as the spherical leucoplast after division, and each half of the dumbbell contained about half the number of nucleoids. Nucleoids increased in number with growth of the leucoplast. The behavior of nucleoids during the leucoplast division cycle in onion bulbs was basically similar to that during the chloroplast division cycle in higher plants and green algae, which was previously reported (Kuroiwa et al. 1981 b).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01283320
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  • 4
    Electronic Resource
    Electronic Resource
    Fluorescence microscopic study of the formation of giant mitochondrial nuclei in the young ovules ofPelargonium zonale (1990)
    Springer
    Protoplasma 158 (1990), S. 191-194 
    Details
    ISSN: 1615-6102
    Keywords: Pelargonium zonale ; Ovale ; Giant mitochondrial nuclei ; DAPI ; Fluorescence microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The size of mitochondrial genomes in higher plants are known to range from 200 to 2400 kilobase pairs. However, we failed to identify cytochemically any mitochondria that contain an identifiable master mitochondrial genome. In the present experiments, we have found the giant mitochondrial nuclei which have the capacity for including the master mitochondrial genome in the young ovaries ofPelargonium zonale by use of a 4′-6-diamidino-2-phenylindole (DAPI) epifluorescence microscopy, a Technovit embedding, and a video-intensified photon counting system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01323132
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  • 5
    Electronic Resource
    Electronic Resource
    Migration of the cell nucleus during the amoebo-flagellate transformation ofPhysarum polycephalum is mediated by an actin-generated force that acts on the centrosome (1991)
    Springer
    Protoplasma 163 (1991), S. 114-124 
    Details
    ISSN: 1615-6102
    Keywords: Physarum polycephalum ; DAPI ; Fluorescence microscopy ; Centrosome ; Comigration ; Centrosome migration ; Cell-nuclear migration ; Actin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mechanism of cell-nuclear migration during the amoebo-flagellate transformation inPhysarum polycephalum was examined by fluorescence microscopy after staining with a tubulinspecific antibody, rhodamine-conjugated phalloidin and 4′,6-diamidino-2-phenylindole (DAPI). While the round amoeba cells changed to comma-shaped swarm cells within 20min after suspension in buffer, the cell nuclei moved from the central region of each cell to the periphery, each forming a sharp projection in the direction of movement. A centrosome also migrated from the center of the cell to the cell periphery. Since the centrosome was in close contact with the tip that protruded from the cell nucleus throughout the cellnuclear migration, the migration of the cell nucleus and the centrosome could be recognized as comigration. Then the flagella began to elongate from the centrosome and the cells became slender and polarized, adopting the so-called “comma-shape”. On the basis of these observations, the transformation process was classified into three steps: cell-nuclear migration, flagella formation and swarm maturation. The comigration of the cell nucleus and the centrosome was not inhibited by the anti-microtubule drug nocodazole (4 μM) but it was inhibited by the anti-microfilament drug cytochalasin A (4 μM), suggesting that the force of migration is generated by microfilaments. To investigate the role of the centrosome in this comigration in detail, we identified two aberrant strains, defective in swimming ability, from among various laboratory strains. The two strains, TM 4 and J, were found to have defects in cell-nuclear migration. Strain TM 4 had two types of irregular swarm cells: in one, only a part of the cell nucleus projected a thin filamentous structure; and in the other, no cell-nuclear migration occurred. Strain J had two centrosomes per cell and such swarm cells exhibited an attempt of cell-nuclear migration at two sites which corresponded to the position of the centrosome. The characteristics of these two strains indicate that the centrosome is essential for cell-nuclear migration. Our observations suggest that the cell-nuclear migration is mediated by actin-generated forces that act on the centrosome rather than on the cell nucleus itself.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01323335
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  • 6
    Electronic Resource
    Electronic Resource
    Absence of DNA in the basal body ofChlamydomonas reinhardtii by fluorimetry using a video-intensified microscope photon-counting system (1990)
    Springer
    Protoplasma 158 (1990), S. 155-164 
    Details
    ISSN: 1615-6102
    Keywords: Chlamydomonas reinhardtii ; DAPI ; Basal bodies ; Absence of DNA ; Fluorimetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A search was made for DNA in both the basal bodies (BBs) in situ and BBs isolated from cells ofChlamydomonas reinhardtii by high-resolution epifluorescence microscopy after staining with 4′-6-diamidino-2-phenylindole (DAPI), by fluorimetry using a video-intensified microscope photon-counting system (VIMPICS) and by immunofluorescence microscopy after staining with a monoclonal tubulin-specific antibody. The flagella and intracellular microtubules radiate from the BBs. The BBs in young vegetative cells, gametes and young zygotes do not emit fluorescence after staining with DAPI but the spherical cell nucleus, the ovoid chloroplast nuclei and the tiny mitochondrial nuclei emit bright, blue-white fluorescence. Thus, it appears that BBs do not contain larger amounts of DNA than do the other organelles. To avoid the halation effects of fluorescence from the cell debris and cytoplasm and to measure carefully any extremely low levels of DNA that might be present in the organelles, a complex, composed of two flagella, a pair of BBs and the cell nucleus, was isolated from the gametes by treatment with autolysin and 0.1% Triton X-100. After staining with DAPI, the BBs of such complexes exhibit faint fluorescence while the cell nucleus emits strong fluorescence. The point and total intensities of the fluorescence emitted from each portion of the complex were measured with the VIMPICS. When the fluorescence intensity “T” of T 4 phage is taken as a standard, the fluorescence intensities of the flagella, the pair of BBs, the cell nucleus and the nucleus ofEscherichia coli are respectively 0.2 T, 0.40 T, 1452.2 T and 20.4 T. The slight fluorescence emitted from the BB seems to be due to the halation of the fluorescence emitted from the cell nucleus. The intensity of the fluorescence from the BBs is reduced to the intensity of the fluorescence of the flagella when the cell nucleus is removed from the complex. From these results, we conclude that the BBs do not contain DNA. Discrepancies related to the reported presence of DNA in the BBs are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01323128
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  • 7
    Electronic Resource
    Electronic Resource
    Behavior of chloroplasts and chloroplast nuclei during spermatogenesis in the fern,Pteris vittata L. (1988)
    Springer
    Protoplasma 146 (1988), S. 89-100 
    Details
    ISSN: 1615-6102
    Keywords: Antheridium ; Antheridiogen ; Chloroplast ; Chloroplast nuclei ; Pteris vittata L. ; Spermatogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The fate of the chloroplasts and chloroplast nuclei (cp-nuclei) was followed during spermatogenesis in the fernPteris vittata L. by epifluorescence microscopy after staining with 4′-6-diamidino-2-phenylindole (DAPI) and by quantitation of chloroplast DNA (cp-DNA) by fluorimetry using a video intensified microscope photon counting system (VIMPICS). The spores were grown on solid medium that contained antheridiogen (Anptd), and formed an antheridium initial on the protonema cell. The antheridium initial divided and produced 16 spermatocytes and 3 surrounding cells. The chloroplasts in the spermatocytes decreased in volume as cell division was repeated, until finally the volume of each chloroplast was 1/15 of that of the primary chloroplasts. The DNA content of the chloroplasts was also reduced to 1/5 of the original value and when the sperm matured, the fluorescence of cp-DNA disappeared. In the 16-cell spermatocyte, the recognition of the fluorescence of chlorophyll in the chloroplasts with a green excitation filter became difficult. But, the plastids could be observed until the final stage of the sperm. From these observations, it appears that there are two steps in the metamorphosis of chloroplasts during spermatogenesis in the fern. The first step involves the decrease in the volume of chloroplasts, accompanied by reduction of the DNA content, and the second step involves the change of the physical state of chloroplasts to amyloplasts and the disappearance of the cp-DNA from the amyloplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01405917
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  • 8
    Electronic Resource
    Electronic Resource
    The course of chloroplast DNA replication and its relationship to other reproductive processes in the chloroplast and nucleocytoplasmic compartment during the cell cycle of the algaScenedesmus quadricauda (1995)
    Springer
    Protoplasma 188 (1995), S. 245-251 
    Details
    ISSN: 1615-6102
    Keywords: Chloroplast DNA ; Chloroplast nuclei ; DAPI ; Mitotic cycle ; Scenedesmus quadricauda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA containing structures (cellular, chloroplast and mitochondrial nuclei) were stained with the fluorochrome DAPI. Fluorescence intensity, as a measure of DNA content, was estimated during the mitotic cycle in synchronized populations of the chlorococcal alga,Scenedesmus quadricauda. In cells yielding eight daughter cells, three consecutive steps in chloroplast DNA increase occurred over one mitotic cycle. The first step was performed shortly after releasing the daughter cells, the second and third steps occurred consecutively during the first half of the mitotic cycle. Commitment to chloroplast DNA replication was chronologically separated from commitment to division of chloroplast nuclei, revealing that these two chloroplast reproductive steps were under different control mechanisms. The replication of chloroplast DNA occurred at a different time to that of cell-nuclear DNA. The coordination of chloroplast reproductive processes and those in the nucleocytoplasmic compartment were governed by the mutual trophic and metabolic dependency of these compartments rather than by any direct or feedback control controlled by either of them.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01280376
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  • 9
    Electronic Resource
    Electronic Resource
    Uncoupling of chloroplast reproductive events from cell cycle division processes by 5-fluorodeoxyuridine in the algaScenedesmus quadricauda (1996)
    Springer
    Protoplasma 192 (1996), S. 228-234 
    Details
    ISSN: 1615-6102
    Keywords: Cell cycle ; Chloroplast cycle ; Chloroplast fission ; Chloroplast nuclei ; Fluorodeoxyuridine ; Scenedesmus quadricauda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary FdUrd (5-fluorodeoxyuridine), a specific inhibitor of thymidylate synthase, was used to study the relationship between reproductive processes in chloroplast and nucleocytoplasmic compartments of the chlorococcal algaScenedesmus quadricauda. The courses of DNA replication and nuclear division in both the compartments were followed in populations synchronised by the alternation of light and dark periods. DAPI-staining of DNA-containing structures was used for their visualisation and quantification. In contrast with cellular reproductive events, those in chloroplasts were not substantially affected by the presence of FdUrd (25 μg/ml). It was shown that FdUrd specifically blocked nucDNA replication but not ptDNA replication. Thus, cells which had attained commitment to ptDNA replication, fission of pt-nuclei and chloroplast kinesis triggered and terminated these processes while the corresponding cellular processes were blocked. The courses of reproductive processes in chloroplasts were also substantially unaffected in cells grown in the presence of FdUrd for the whole cell cycle. This provided evidence that attainment of commitment to and termination of the entire sequence of reproductive events, including chloroplast fission, were controlled by different mechanisms than the reproductive processes in the nucleocytoplasmic compartment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01273894
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  • 10
    Electronic Resource
    Electronic Resource
    Preferential degradation of plastid DNA with preservation of mitochondrial DNA in the sperm cells ofPelargonium zonale during pollen development (1997)
    Springer
    Protoplasma 197 (1997), S. 217-229 
    Details
    ISSN: 1615-6102
    Keywords: Biparental inheritance ; Fluorescence microscopy ; Immunoelectron microscopy ; Maternal inheritance ; Pelargonium zonale ; Pollen grains
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the present study, we studied changes in organellar DNA in the sperm cells of maturing pollen ofPelargonium zonale, a plant typical to exhibit biparental inheritance, by fluorescence microscopy after staining with 4′,6-diamidino-2-phenylindole (DAPI) and by immunogold electron microscopy using anti-DNA antibody. Fluorescence intensities of DAPI-stained plastid nuclei in generative and sperm cells at various developmental stages were quantified with a video-intensified microscope photon counting system (VIMPCS). Results indicated that the amount of DNA per plastid in generative cells increased gradually during pollen development and reached a maximum value (about 70 T per plastid; 1 T represents the amount of DNA in a particle of T4 phage) in young sperm cells at 5 days before flowering. However, the DNA content of plastids was subsequently reduced to about 20% of the maximum value on the day of flowering. Moreover, the DNA content of the plastid further decreased to 4% of the maximum value when pollen grains were cultured for 6 h in germination medium. In contrast, the amount of DNA per mitochondrion did not decrease significantly around the flowering day. Similar results were also obtained by immunogold electron microscopy using anti-DNA antibody. The density of gold particles on plastids decreased during pollen maturation whereas labelling density on mitochondria remained relatively constant. The number of plastids and mitochondria per generative cell or per pair of sperm cells did not change significantly, indicating that the segregation of DNA by plastid division was not responsible for the decrease in the amount of DNA per plastid. These results indicate that the plastid DNA is preferentially degraded, but the mitochondrial DNA is preserved, in the sperm cells ofP. zonale. While the plastid DNA of the sperm cells decreased before fertilization, it was also suggested that the low DNA contents that remain in the plastids of the sperm cells are enough to account for the biparental inheritance of plastids inP. zonale.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01288031
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