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Cloning and functional characterization of plant pathogen inducible promoters is of great significance for their use in the effective management of plant diseases. The rice gene  CYP76M7  was up regulated at 24, 48, and 72 hours post inoculation (hpi) with two isolates of  Magnaporthe oryzae  Mo-ei-11 and Mo-ni-25. In this study, the promoter of  CYP76M7  gene was cloned from rice cultivar HR-12, characterized and functionally validated. The Transcription Start Site of  CYP76M7  was mapped at 45 bases upstream of the initiation codon. To functionally validate the promoter, 5′ deletion analysis of the promoter sequences was performed and the deletion fragments fused with the β -glucuronidase (GUS)  reporter gene were used for generating stable transgenic  Arabidopsis  plants as well as for transient expression in rice. The spatial and temporal expression pattern of GUS in transgenic  Arabidopsis  plants and also in transiently expressed rice leaves revealed that the promoter of  CYP76M7  gene was induced by  M. oryzae . The induction of  CYP76M7  promoter was observed at 24 hpi with  M. oryzae . We report that, sequences spanning -222 bp to -520 bp, with the cluster of three W-boxes, two ASF1 motifs and a single GT-1 element may contribute to the  M. oryzae  inducible nature of  CYP76M7  promoter. The promoter characterized in this study would be an ideal candidate for the overexpression of defense genes in rice for developing durable blast resistance rice lines.

Cloning and functional validation of early inducible Magnaporthe oryzae responsive CYP76M7 promoter from rice