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In this study, the differences in reactive oxygen species (ROS) generation and abscisic acid (ABA) accumulation in senescing leaves were investigated by early-senescence-leaf ( esl ) mutant and its wild type, to clarify the relationship among ABA levels, ROS generation, and NADPH oxidase (Nox) in senescing leaves of rice ( Oryza sativa ). The temporal expression levels of  OsNox  isoforms in senescing leaves and their expression patterns in response to ABA treatment were determined through quantitative real-time reverse transcription PCR (qRT-PCR). Results showed that the flag leaf of the  esl  mutant generated more O 2 -   concentrations and accumulated higher ABA levels than the wild-type cultivar did in the grain-filling stage. Exogenous ABA treatment induced O 2 -   generation; however, it was depressed by diphenyleneiodonium chloride (DPI) pretreatment in the detached leaf segments. This finding suggested the involvement of NADPH oxidase in ABA-induced O 2 -   generation. The  esl  mutant exhibited significantly higher expression of  OsNox2 ,  OsNox5 ,  OsNox6 , and  OsNox7  in the initial of grain-filling stage, followed by sharply decrease. The transcriptional levels of  OsNox1 ,  OsNox3 , and  OsFR07  in the flag leaf of the  esl  mutant were significantly lower than those in the wild-type cultivar. The expression levels of  OsNox2 ,  OsNox5 ,  OsNox6 , and  OsNox7  were significantly enhanced by exogenous ABA treatments. The enhanced expression levels of  OsNox2  and  OsNox6  were dependent on the duration of ABA treatment. The inducible expression levels of  OsNox5  and  OsNox7  were dependent on ABA concentrations. By contrast, exogenous ABA treatment severely repressed the transcripts of  OsNox1 ,  OsNox3 , and  OsFR07  in the detached leaf segments. Therefore,  OsNox2 ,  OsNox5 ,  OsNox6 , and  OsNox7  were probably involved in the ABA-induced O 2 -   generation in the initial stage of leaf senescence. Subsequently, other oxidases activated in deteriorating cells were associated with ROS generation and accumulation in the senescing leaves of the  esl  mutant. Conversely,  OsNox1 ,  OsNox3 , and  OsFR07  were not associated with ABA-induced O 2 -   generation during leaf senescence.

plos one

Involvement of NADPH oxidase isoforms in the production of O2− manipulated by ABA in the senescing leaves of early-senescence-leaf (esl) mutant rice (Oryza sativa)