Summary
Cytoskeletal organization and chromosome behavior were studied inTradescantia generative cells prior to and during sperm formation using in vitro grown pollen tubes and fluorescence staining methods. Before pollen germination, the crescent-shaped generative cell contains a reticulate microtubule (Mt) system. The cell elongates dramatically after germination, and its Mts assume a helical to longitudinal arrangement. Chromosome condensation is evident approximately 3hr after germination. Kinetochores appear as dark interruptions in the Mt array, and thus seem to attach directly to interphase fibers. No metaphase plate typical of other cells is observed with either DAPI or anti-tubulin staining. Instead, the chromosomes adopt a twisted or braided arrangement, with kinetochores distributed along the length of the cell and kinetochore fibers linked to each other and to surrounding fibers. Anaphase is characterized by a staggered, overlapping separation of chromosomes and by elongation of Mt branches connecting opposing kinetochore fibers. Cytokinesis appears to utilize a furrowing process; a phragmoplast or cell plate was never seen. As a result of these events, the sperm directly inherit their cytoskeleton from generative cell Mts involved in division. No actin fibers are observed at any stage using rhodamine-phalloidin staining. The results are discussed in terms of other reports on sperm formation, possible mitotic and cytokinetic mechanisms, and past distinctions between Mt arrays in higher plant somatic cells.
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Abbreviations
- CD:
-
cytochalasin D
- DAPI:
-
4′6-diamidino-2-phenyl-indole
- DMSO:
-
dimethylsulfoxide
- K-fiber:
-
kinetochore fiber
- Mf:
-
microfilament
- Mt:
-
microtubule
- PPB:
-
preprophase Mt band
- RP:
-
rhodamine phalloidin
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Palevitz, B.A., Cresti, M. Cytoskeletal changes during generative cell division and sperm formation inTradescantia virginiana . Protoplasma 150, 54–71 (1989). https://doi.org/10.1007/BF01352921
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DOI: https://doi.org/10.1007/BF01352921