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Keywords:

  • apoptosis;
  • Brassica napus;
  • necrosis;
  • programmed cell death;
  • protoplast

Summary

  • •  
    The cleavage of nuclear DNA into oligonucleosomal fragments that is the hallmark of apoptosis in animal cells occurs during the culture of Brassica napus leaf protoplasts.
  • •  
    The changes in nuclei of cultured Brassica napus leaf protoplasts were studied by propidium iodide (PI) and 4′, 6-diamino-2-phenylindole, dihydrochloride (DAPI) staining, transmission electron microscopy, flow cytometry analysis, and DNA laddering staining with ethidium bromide and Southern hybridization.
  • •  
    Free 3′-OH termini of nuclear DNA fragments were labelled with DIG-dUTP, catalyzed by terminal deoxynucleotidyl transferase (TdT), and used as probes for Southern hybridization. This method (TUNEL on membrane) allowed visualization of DNA fragments with 3′-OH termini on a nylon membrane.
  • •  
    These results suggest that loss of viability of protoplast with culture time is accompanied by apoptosis-like cell death. However, the forms or processes undergoing to apoptotic cell death in B. napus leaf protoplasts appears to be different in some details to those in animal cells.