Apoptosis in suspension culture of tobacco cells induced by heat shock
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The induction of apoptosis in suspension culture of tobacco cells by heat shock is reported for the first time. Heat treatment (48° for 4 h) of tobacco cells led to the appearance of typical hallmarks of apoptosis. It was demonstrated by DNA laddering analysis that the cells treated with heat shock at 48°C for 4 h had a serious degradation of nuclear DNA into multi-nucleosomal sizes, suggesting that heat shock activated endogenous nuclease which led to DNA cleavage at the linkage sites between the nucleosomes, but ladders were very faint for DNA from2 and 9 h heat-treated cells. The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) detection also showed that most of these treated cells (48°C for4 h) displayed positive reactions, indicating a serious DNA 3′-OH cleavage in their nuclei. Moreover, some other cytological changes in apoptotic cells, such as cell shrinkage, chromatin aggregation, nucleus collapse, have also been observed by 4′, 6′-diamidino-2-phenyl-indole (DAPI) staining.
Keywordsapoptosis DNA ladder TUNEL Nicotiana tabacum
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