Apoptosis-like cell death of Brassica napus leaf protoplasts
Article first published online: 24 NOV 2002
Volume 156, Issue 3, pages 417–426, December 2002
How to Cite
Watanabe, M., Setoguchi, D., Uehara, K., Ohtsuka, W. and Watanabe, Y. (2002), Apoptosis-like cell death of Brassica napus leaf protoplasts. New Phytologist, 156: 417–426. doi: 10.1046/j.1469-8137.2000.00536.x
- Issue published online: 24 NOV 2002
- Article first published online: 24 NOV 2002
- Received: 23 June 2002 Accepted: 9 September 2002
- Brassica napus;
- programmed cell death;
- • 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.