Independent pathways leading to apoptotic cell death, oxidative burst and defense gene expression in response to elicitin in tobacco cell suspension culture.
We characterized pharmacologically the hypersensitive cell death of tobacco BY-2 cells that followed treatments with Escherichia coli preparations of INF1, the major secreted elicitin of the late blight pathogen Phytophthora infestans. INF1 elicitin treatments resulted in fragmentation and 180 bp laddering of tobacco DNA as early as 3 h post-treatment. INF1 elicitin also induced rapid accumulation of H2O2 typical of oxidative burst, and the expression of defense genes such as phenylalanine ammonia-lyase (PAL) gene at 1 h and 3 h after elicitin treatment, respectively. To investigate the involvement of the oxidative burst and/or the expression of defense genes in the signal transduction pathways leading to hypersensitive cell death, we analyzed the effect of several chemical inhibitors of signal transduction pathways on the various responses. The results indicated that (a) the cell death required serine proteases, Ca2+ and protein kinases, (b) the oxidative burst was involved in Ca2+ and protein kinase mediated pathways, but elicitin-induced AOS was neither necessary nor sufficient for cell death and PAL gene expression, and (c) the signaling pathway of PAL gene expression required protein kinases. These results suggest that the three signal transduction pathways leading to cell death, oxidative burst and expression of defense genes branch in the early stages that follow elicitin recognition by tobacco cells.
Science and Technology for Energy Conversion, Graduate School of Natural Science and Technology, and Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University, Japan.
M
M
Sasabe
K
K
Takeuchi
S
S
Kamoun
Y
Y
Ichinose
F
F
Govers
K
K
Toyoda
T
T
Shiraishi
T
T
Yamada
D020418:Algal Proteins; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D004926:Escherichia coli; D005656:Fungal Proteins / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D007700:Kinetics; D010650:Phenylalanine Ammonia-Lyase / Q000235:genetics; D010838:Phytophthora; D010947:Plants, Toxic; D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins; D017382:Reactive Oxygen Species / Q000502:physiology; D011994:Recombinant Proteins / Q000494:pharmacology; D016897:Respiratory Burst / Q000187:drug effects / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014026:Nicotiana / Q000166:cytology* / Q000235:genetics / Q000502:physiology*
D020418:Algal Proteins; D005656:Fungal Proteins; D011506:Proteins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; C085101:elicitin, Phytophthora; D011494:Protein Kinases; D010650:Phenylalanine Ammonia-Lyase