Bcl-2 suppresses hydrogen peroxide-induced programmed cell death via OsVPE2 and OsVPE3, but not via OsVPE1 and OsVPE4, in rice

Hydrogen peroxide (H2O2) is known to be a key player in apoptosis in animals. The components and pathways regulating H2O2-induced programmed cell death in plants, however, remain largely unknown. In the present study, rice transgenic lines overexpressing Bcl-2, a human apoptotic suppressor, were obtained. These transgenic lines showed increased tolerance to high levels of H2O2, resulting in increased seed germination rates, root elongation, root tip cell viability and chlorophyll retention compared to control lines. In the control lines, treatment with H2O2 resulted in DNA laddering and a clear terminal transferase dUTP nick end labeling signal, which are the hallmarks of programmed cell death. However, this effect was not detected in the Bcl-2-overexpressing transgenic lines. Further investigations indicated that Bcl-2 suppressed H2O2-induced programmed cell death but did not inhibit stress-elicited reactive oxygen species production in rice. RT-PCR revealed that the expression of the two vacuolar processing enzyme genes (i.e. OsVPE2 and OsVPE3) was dramatically induced by H2O2 in the wild-type line but not in the Bcl-2-overexpressing line. Moreover, treatment with H2O2 resulted in the disruption of the vacuolar membrane in the wild-type line. The expression levels of OsVPE1 and OsVPE4 did not significantly differ between the wild-type line and the transgenic line that was treated or untreated with H2O2. The similar roles of Bcl-2 and OsVPEs during endogenous reactive oxygen species-triggered programmed cell death were also confirmed by NaCl stress in rice. To our knowledge, the present study is the first to demonsatrate that Bcl-2 overexpression inhibits H2O2-induced programmed cell death and enhances H2O2 tolerance. We propose that Bcl-2 overexpression in rice suppresses the transcriptional activation of OsVPE2 and OsVPE3, but not of OsVPE1 or OsVPE4.