Microtubular organization during asymmetrical division of the generative cell inGagea lutea View Full Text


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Article Info

DATE

1995-09

AUTHORS

Hong-Qi Zhang, Jerzy Bohdanowicz, Elisabeth S. Pierson, Yi-Qin Li, Antonio Tiezzi, Mauro Cresti

ABSTRACT

Video microscopy and conventional or Confocal Laser Scanning Microscopy after DAPI staining and anti-α-tubulin labelling were used to study the asymmetrical division of the generative cell (GC) inGagea lutea. Pollen was cultured for up to 8 hr in a medium containing 10% poly (ethylene glycol), 3.0% to 3.8% sucrose, 0.03% casein acid hydrolysate, 15 mM 2-(N-morpholinoethane)-sulphonic acid-KOH buffer (pH 5.9) and salts. In the pollen grain, the GC had a spherical or ovoid shape and contained a fine network of intermingled microtubules. As the GC entered into the pollen tube, it assumed a cylindrical shape with a length often exceeding 250 μm. A cage of microtubules then developed around the nucleus. The presence of dense and thick microtubular bundles in front of the generative nucleus within the GC coincided with the translocation of the nucleus to the leading end of the GC. No pre-prophase band was ever detected, but a distinct prophase spindle of microtubules was formed. In some GCs a tubulin-rich dot became visible at each pole of the spindle. After nuclear envelope breakdown, the bundles of microtubules spread between the chromosomes and became oriented into parallel arrays. The spindle became shorter at metaphase, and there was no tubulin labelling at the site of the metaphase plate. At anaphase, the microtubular apparatus lost its spindle-shape and a bridge of prominent bundles of microtubules connected the two daughter nuclei. At telophase, the site of the cell plate remained unstained by the anti-α-tubulin antibody, but a distinct phragmoplast of microtubules was formed more closely to the leading nucleus, resulting in the formation of unequal sperm cells (SCs). The leading SC was up to 2.5 times smaller than the following SC and it contained a smaller or equal number of nucleoli. More... »

PAGES

269-276

References to SciGraph publications

  • 1987-02. Ultrastructural analysis of the sperm cells of mature pollen of maize,Zea mays in PROTOPLASMA
  • 1984-11. Ultrastructure of the sperm of Plumbago zeylanica in PLANTA
  • 1991-04. Mitosis and microtubule organizational changes in isolated generative cells of Allemanda neriifolia in PLANT REPRODUCTION
  • 1985-10. The spatial association of the sperm cells and vegetative nucleus in the pollen grain ofBrassica in PROTOPLASMA
  • 1993-09. Ontogenesis of monad pollen inPterostylis plumosa (Orchidaceae, Neottioideae) in PLANT SYSTEMATICS AND EVOLUTION
  • 1993-09. Requirements for division of the generative nucleus in cultured pollen tubes ofNicotiana in PROTOPLASMA
  • 1988. The Male Germ Unit of Zea mays: Quantitative Ultrastructure and Three-Dimensional Analysis in SEXUAL REPRODUCTION IN HIGHER PLANTS
  • 1991-06. Cold-stability in the pollen tube cytoskeleton in PROTOPLASMA
  • 1992-03. The organization of the cytoskeleton in the generative cell and sperms ofHyacinthus orientalis in PROTOPLASMA
  • 1994-07. γ-Tubulin in tobacco pollen tubes: association with generative cell and vegetative microtubules in PLANT REPRODUCTION
  • 1989-11. The male germ unit of Rhododendron: quantitative cytology, three-dimensional reconstruction, isolation and detection using fluorescent probes in PLANT REPRODUCTION
  • 1992-03. The microtubule cytoskeleton and the rounding of isolated generative cells ofNicotiana tabacum in PLANT REPRODUCTION
  • 1987-04. Quantitative cytology of the sperm cells of Brassica campestris and B. oleracea in PLANTA
  • 1985-02. Computer-assisted reconstruction of the male germ unit in pollen ofBrassica campestris in PROTOPLASMA
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    DOI

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    39 schema:description Video microscopy and conventional or Confocal Laser Scanning Microscopy after DAPI staining and anti-α-tubulin labelling were used to study the asymmetrical division of the generative cell (GC) inGagea lutea. Pollen was cultured for up to 8 hr in a medium containing 10% poly (ethylene glycol), 3.0% to 3.8% sucrose, 0.03% casein acid hydrolysate, 15 mM 2-(N-morpholinoethane)-sulphonic acid-KOH buffer (pH 5.9) and salts. In the pollen grain, the GC had a spherical or ovoid shape and contained a fine network of intermingled microtubules. As the GC entered into the pollen tube, it assumed a cylindrical shape with a length often exceeding 250 μm. A cage of microtubules then developed around the nucleus. The presence of dense and thick microtubular bundles in front of the generative nucleus within the GC coincided with the translocation of the nucleus to the leading end of the GC. No pre-prophase band was ever detected, but a distinct prophase spindle of microtubules was formed. In some GCs a tubulin-rich dot became visible at each pole of the spindle. After nuclear envelope breakdown, the bundles of microtubules spread between the chromosomes and became oriented into parallel arrays. The spindle became shorter at metaphase, and there was no tubulin labelling at the site of the metaphase plate. At anaphase, the microtubular apparatus lost its spindle-shape and a bridge of prominent bundles of microtubules connected the two daughter nuclei. At telophase, the site of the cell plate remained unstained by the anti-α-tubulin antibody, but a distinct phragmoplast of microtubules was formed more closely to the leading nucleus, resulting in the formation of unequal sperm cells (SCs). The leading SC was up to 2.5 times smaller than the following SC and it contained a smaller or equal number of nucleoli.
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